Antioxidants doi: 10.3390/antiox14010091

Authors: Hongjin Zhou Zhongxiong Fan Yuan Li Xuelian Liu Bo Wang Jianguo Xing Jiang He Ruifang Zheng Jinyao Li

To investigate the structure–antioxidant activity relationship, Pleurotus ferulae polysaccharides were extracted using ultrasonic (U-PFPS) and microwave/ultrasonic-assisted methods (MU-PFPS). Compared to U-PFPS with a molecular weight of 1.566 × 103 kDa, MU-PFPS exhibited a lower molecular weight of 89.26 kDa. In addition, unlike U-PFPS, which is primarily composed of glucose (Glu:Man:Gal = 91.1:3.5:5.4), MU-PFPS has a more balanced composition of Glu:Man:Gal in the ratio of 39.4:27.8:32.8 and contains more branched chains. Furthermore, antioxidant analysis revealed that high concentration (at concentrations above 600 μg/mL) MU-PFPS demonstrated stronger protective effects against oxidative damage in RAW264.7 cells than U-PFPS did. Collectively, these data suggest that lower molecular weight and higher branching degree of polysaccharides at appropriate concentrations may correlate with enhanced antioxidant enzyme activities. Our work provides a method for isolating polysaccharides with higher antioxidant activity and offers insights into the structure–activity relationship of polysaccharides, laying the foundation for future applications in polysaccharide modification and structural characterization.

Antioxidants doi: 10.3390/antiox14010090

Authors: Robert J. Aitken Alexandra Wilkins Natasha Harrison Mohammad Bahrami Zamira Gibb Kaitlin McIntosh Quan Vuong Sarah Lambourne

(1) Background: The RoXstaTM system has been developed as a rapid, effective means of profiling different types of antioxidant activity. The purpose of this study was to examine its performance utilizing a diverse array of biological fluids including semen, blood plasma, serum, urine, saliva, follicular fluid and plant extracts. (2) Methods: The RoXstaTM system was used to assess the ability of different fluids to suppress free radical formation as well as scavenge a variety of toxic oxygen metabolites including free radicals and both hydrogen and organic peroxides. (3) Results: Human semen was shown to have significantly (p < 0.001) more peroxide scavenging power than any other fluid tested (10–14 mM vitamin C equivalent compared with 1–2 mM for blood serum or plasma), while urine was particularly effective in scavenging free radicals and preventing free radical formation (p < 0.001). The powerful antioxidant properties of human semen were shown to reside within the seminal plasma (SP) fraction, rather than the spermatozoa, and to be resistant to snap freezing in liquid nitrogen. Moreover, comparative studies demonstrated that human SP exhibited significantly (p < 0.001) higher levels of antioxidant potential than any other species examined (stallion, bull, dog) and that this intense activity reflected the relative vulnerability of human spermatozoa to peroxide attack. (4) Conclusions: The RoXstaTM system provides valuable information on the antioxidant profile of complex biological fluids, supporting its diagnostic role in conditions associated with oxidative stress. Based on the results secured in this study, human semen is identified as a particularly rich source of antioxidants capable of scavenging both hydrogen and organic peroxides, in keeping with the high susceptibility of human spermatozoa to peroxide-mediated damage.

Antioxidants doi: 10.3390/antiox14010089

Authors: Ioan Pet Igori Balta Nicolae Corcionivoschi Tiberiu Iancu Ducu Stef Lavinia Stef Iuliana Cretescu

The absence of efficient on-farm interventions against white spot syndrome viral (WSSV) infections can cause significant economic losses to shrimp farmers. With this exploratory study we aimed to test, both in vitro and in vivo, the efficacy of an organic acid mixture (Aq) against WSSV infections in shrimp. In vitro, using shrimp gut primary cells (SGP), 2% Aq significantly reduced WSSV infection and the amounts of H2O2 released but had no impact on CAT and SOD expression. In vivo, in a shrimp challenge test, 2% Aq significantly downregulated the expression of proteins involved in WSSV virulence, such as the lipopolysaccharide-β-1,3-glucan-binding protein (LGBP) and the TLR signalling pathway (LvECSIT), and increased the expression of HO-1 oxygenase. Additionally, at 2% Aq, the expression of the digestive-related enzyme carboxypeptidase B was upregulated in the gut, alongside a significant decrease in IL-22 expression, a cytokine usually increased during WSSV infection in shrimp. A low mortality rate (7.33%) was recorded in infected shrimp treated with 2% Aq compared to the 96.66% mortality in the absence of Aq. The peritrophic membrane (PM) was proven essential to ensure Aq efficacy, as the infected and treated PM deficient shrimp (PM−) had a mortality rate of 27.8%, compared to only 9.34% mortality in the infected shrimp at 2% Aq and in the presence of PM (PM+). Aq significantly increased the expression of mucin-1, mucin-2, mucin-5AC, mucin-5B, and mucin-19 in both PM+ and PM− shrimp. Conclusively, organic acid in mixtures can protect farmed shrimp against WSSV infection and increase their survivability through a mediated gut health effect which includes resistance to oxidative stress and improved immunity.

Antioxidants doi: 10.3390/antiox14010088

Authors: Debora Dessì Giacomo Fais Paolo Follesa Giorgia Sarais

The rising global focus on healthy lifestyles and environmental sustainability has prompted interest in repurposing plant-based by-products for health benefits. With increasing life expectancy, the incidence of neurodegenerative diseases—characterized by complex, multifactorial mechanisms such as abnormal protein aggregation, mitochondrial dysfunction, oxidative stress, and inflammation—continues to grow. Medicinal plants, with their diverse bioactive compounds, offer promising therapeutic avenues for such conditions. Myrtus communis L., a Mediterranean plant primarily used in liquor production, generates significant waste rich in antioxidant and anti-inflammatory properties. This study explores the neuroprotective potential of Myrtus berry by-products in a cellular model of neurodegeneration. Using PC12 cells exposed to 6-hydroxydopamine (6-OHDA), we assessed cell viability via MTT assay and measured reactive oxygen species (ROS) production using DCFDA fluorescence. Additionally, we analyzed the expression of genes linked to oxidative stress and neuronal function, including AChE, PON2, Grin1, Gabrd, and c-fos, by RT-PCR. Our findings reveal that Myrtus extract significantly protects against 6-OHDA-induced cytotoxicity, reduces ROS levels, and modulates the expression of key stress-related genes, underscoring its potential as a neuroprotective agent. These results highlight the therapeutic promise of Myrtus extracts in mitigating neurodegenerative processes, paving the way for future interventions.

Antioxidants doi: 10.3390/antiox14010087

Authors: Xiao Zhou Li Wang Jiuli Dai Huiyan Jia Kai Shi Jian Zhao Shufang Chen

Spartina anglica (SA), a plant rich in dietary fiber, has demonstrated considerable potential for enhancing gut health and antioxidant capacity in animals. This study investigates the integration of SA as a novel dietary ingredient for Zhedong white geese, with a specific focus on evaluating its effects on growth performance, nutrient digestibility, antioxidant capacity, intestinal health, and cecal microbiota composition. A total of 360 1-day-old Zhedong white geese with an average weight of 114.94 ± 0.81 g were randomly allocated to 4 dietary treatments, with 6 replicates per treatment and 15 geese per pen. The 4 dietary treatments included different SA supplement levels: a control group receiving a basal diet (CON), and three experimental groups supplemented with 3% SA (SA3), 6% SA (SA6), and 12% SA (SA12). Supplementation with 6% SA significantly enhanced the final body weight, average daily gain, and feed conversion ratio (FCR) compared to the CON group (p < 0.05). In contrast, the SA12 group exhibited reduced digestibility of crude protein and ether extract, relative to the SA3 and SA6 groups (p < 0.05). The highest antioxidant capacity was observed in the SA6 and SA12 groups, while the lowest was recorded in the CON group. SA supplementation did not significantly influence serum biochemical parameters or organ indices but increased cecum length (p < 0.05). Notably, SA supplementation markedly improved intestinal morphology, although excessive levels appeared to compromise these benefits. Additionally, SA supplementation significantly enhanced the richness and diversity of cecal microbiota and increased short-chain fatty acid concentrations. In conclusion, SA at an optimal supplementation level of 6% may be effectively utilized in Zhedong white geese diets to improve growth performance, gut health, and antioxidant capacity.

Antioxidants doi: 10.3390/antiox14010085

Authors: Bin Du Baojun Xu

Oxidative stress is caused by various intrinsic and extrinsic factors [...]

Antioxidants doi: 10.3390/antiox14010086

Authors: Natalia Adamczuk Mirosława Krauze-Baranowska Justyna Ośko Małgorzata Grembecka Piotr Migas

The aim of this study was to compare the antioxidant potential in the fruits of different hybrids of Rubus idaeus and Rubus occidentalis (four hybrids) against the fruit of known cultivars of both species (R. idaeus—three cultivars; R. occidentalis—five cultivars) and, using chemometric analysis, to select factors affecting the level of polyphenols and antioxidant properties. Antioxidant activity was determined using the ABTS, DPPH and FRAP tests. Chemometric analysis enabled the separation of R. idaeus and R. occidentalis cultivars and classified the hybrid R. idaeus/R. occidentalis R1314701 as belonging to the R. occidentalis species. Moreover, two hybrids, Rubus occidentalis/Rubus idaeus R1613411 and R. idaeus/R. occidentalis R1613409, can be classified as a purple raspberry. Crossbreeding species/cultivars of the Rubus genus may result in an increased content of anthocyanins, but on the other hand, it may lead to a reduction in free radical scavenging activity in the ABTS and DPPH. Spearman’s correlations confirm the correlations between the total polyphenol content and antioxidant activity in the DPPH, ABTS and FRAP, as well as the anthocyanin content and antioxidant activity in the ABTS and FRAP tests. Chemometric analysis can be an effective tool in determining the species affiliation of obtained hybrids and cultivars.

Antioxidants doi: 10.3390/antiox14010084

Authors: Francisco Aguirre Franco Tacchi Mayalen Valero-Breton Josué Orozco-Aguilar Sabrina Conejeros-Lillo Josefa Bonicioli Renata Iturriaga-Jofré Daniel Cabrera Jorge A. Soto Mauricio Castro-Sepúlveda Marianny Portal-Rodríguez Álvaro A. Elorza Andrea Matamoros Felipe Simon Claudio Cabello-Verrugio

Sarcopenia corresponds to a decrease in muscle mass and strength. CCL5 is a new myokine whose expression, along with the CCR5 receptor, is increased in sarcopenic muscle. Therefore, we evaluated whether CCL5 and CCR5 induce a sarcopenic-like effect on skeletal muscle tissue and cultured muscle cells. Electroporation in the tibialis anterior (TA) muscle of mice was used to overexpress CCL5. The TA muscles were analyzed by measuring the fiber diameter, the content of sarcomeric proteins, and the gene expression of E3-ligases. C2C12 myotubes and single-isolated flexor digitorum brevis (FDB) fibers were also treated with recombinant CCL5 (rCCL5). The participation of CCR5 was evaluated using the antagonist maraviroc (MVC). Protein and structural analyses were performed. The results showed that TA overexpression of CCL5 led to sarcopenia by reducing muscle strength and mass, muscle-fiber diameter, and sarcomeric protein content, and by upregulating E3-ligases. The same sarcopenic phenotype was observed in myotubes and FDB fibers. We showed increased reactive oxygen species (ROS) production and carbonylated proteins, denoting oxidative stress induced by CCL5. When the CCR5 was antagonized, the effects produced by rCCL5 were prevented. In conclusion, we report for the first time that CCL5 is a novel myokine that exerts a sarcopenic-like effect through the CCR5 receptor.

Antioxidants doi: 10.3390/antiox14010083

Authors: Vittoria Cammisotto Emanuele Valeriani Pasquale Pignatelli Francesco Violi

Metabolic dysfunction-associated steatotic liver disease (MASLD) is characterized by lipid accumulation in the liver due to an excess in their supplies or an impairment in their management. While some patients remain stable for years, a proportion of them progress up to steatohepatitis (MASH). MASLD links with systemic pathways being associated with metabolic and non-metabolic diseases. Although liver lipid accumulation represents the first hit for MASLD, the pathophysiology of its development and progression to MASH remains not completely understood. Oxidative stress has received particular attention in recent years, as most of the oxidative process occurs in the liver, which is also the target of oxidative stress-induced damage. Growing evidence linked the activity of nicotinamide adenine dinucleotide phosphate (NADPH) oxidases (NOX) to the increased liver production of reactive oxygen species up to liver damage and fibrosis. NOX acts both in hepatocytes and in non-parenchymal hepatic cells, contributing to hepatocyte lipotoxicity, impaired hepatic microcirculation, hepatic stellate, and mesenchymal stem cells activation and proliferation. This review aims to summarize the current knowledge on the involvement of oxidative stress in the MASLD–MASH transition, focusing on the role of NOX isoforms, and to suggest targeting NOX as a therapeutic approach in MASLD.

Antioxidants doi: 10.3390/antiox14010082

Authors: Jessica Cusato Anna Mulasso Micol Ferrara Alessandra Manca Guido Accardo Alice Palermiti Miriam Antonucci Gianluca Bianco Francesco Chiara Jacopo Mula Maria Cristina Tettoni Simone Cuomo Giulia Trevisan Stefano Bonora Giovanni Di Perri Corrado Lupo Alberto Rainoldi Antonio D’Avolio

High levels of reactive oxygen species (ROS) are present in people living with HIV (PLWH), produced by intense physical activity; in response, our body produces antioxidant molecules. ROS influence the expression of gene-encoding enzymes and transporters involved in drug biotransformation. In addition, pharmacogenetics can influence transporter activity, and thus drug exposure. Currently, no studies concerning this topic are present in the literature. The aim of this study was to investigate whether some antioxidant molecules, physical exercise, and genetic variants could affect dolutegravir (DTG) concentrations in PLWH, switching from triple to dual therapy. Thirty PLWH were recruited and analyzed at baseline (triple therapy), and 6 months after (dual therapy). Physical capacities were investigated using validated tools. Drug concentrations and oxidative stress biomarkers levels were evaluated through liquid chromatography coupled with tandem mass spectrometry, while genetic variants through real-time PCR. No statistical differences were suggested for drug concentrations, with the exception of intracellular DTG (p = 0.047). Statistically significant correlations between DTG plasma concentrations and white blood cells (p = 0.011; S = 0.480) and cytoplasmic N-acetyl-cysteine (p = 0.033; S = −0.419) were observed. Finally, white blood cells and BMI remained in the final multivariate regression model as predictors of DTG concentrations. This is the first study showing possible factors related to oxidative stress impacting DTG exposure.

Antioxidants doi: 10.3390/antiox14010081

Authors: Vanessa Bertolucci André Felipe Ninomiya Giovanna Barbarini Longato Luisa Oliveira Kaneko Nilson Nonose Pedro Paulo Menezes Scariot Leonardo Henrique Dalcheco Messias

This narrative review explores the potential effects of Propolis and its bioactive compounds on bone health. Propolis, a resinous product collected by bees, is renowned for its antimicrobial, anti-inflammatory, and antioxidant properties. Recent research emphasizes its positive role in osteogenesis, primarily through the modulation of osteoclast and osteoblast activity via molecular pathways. Key mechanisms include reducing inflammatory cytokines, protecting against oxidative stress, and upregulating growth factor essential for bone formation. While compounds such as Caffeic Acid Phenethyl Ester, Apigenin, Quercetin, and Ferulic Acid have been well-documented, emerging evidence points to the significant roles of less-studied compounds like Pinocembrin, Kaempferol, p-Coumaric acid, and Galangin. This review synthesizes the current literature, focusing on the mechanisms by which these bioactive compounds influence osteogenesis. Firstly, it explores the techniques for characterizing bioactive compounds presented in propolis, the chemogeographic variations in its composition, and the effects of both crude extracts and isolated compounds on bone tissue, offering a comprehensive analysis of recent findings across different experimental models. Further, it discusses the effects of Propolis compounds on bone health. In summary, these compounds modulate signaling pathways, including nuclear factor kappa beta, wingless-related integration site, mitogen-activated protein kinase, vascular endothelial growth factor, and reactive oxygen species. These pathways influence the receptor activator of nuclear factor kappa-β/receptor activator of nuclear factor kappa-β ligand/osteoprotegerin system, fostering bone cell differentiation. This regulation mitigates excessive osteoclast formation, stimulates osteoblast activity, and ultimately contributes to the restoration of bone homeostasis by maintaining a balanced bone remodeling process.

Antioxidants doi: 10.3390/antiox14010080

Authors: Francesco Abbiati Ivan Orlandi Stefania Pagliari Luca Campone Marina Vai

Glucosinolates (GSLs) are nitrogen/sulfur-containing glycosides widely present in the order of Brassicales, particularly in the Brassicaceae family. Camelina (Camelina sativa (L.) Crantz) is an oilseed plant belonging to this family. Its seeds, in addition to a distinctive fatty acid composition, contain three aliphatic GSLs: glucoarabin, glucocamelinin, and homoglucocamelinin. Our study explored the impact of these GSLs purified from Camelina press cake, a by-product of Camelina oil production, on yeast chronological aging, which is the established model for simulating the aging of post-mitotic quiescent mammalian cells. Supplementing yeast cells with GSLs extends the chronological lifespan (CLS) in a dose-dependent manner. This enhancement relies on an improved mitochondrial respiration efficiency, resulting in a drastic decrease of superoxide anion levels and an increase in ATP production. Furthermore, GSL supplementation affects carbon metabolism. In particular, GSLs support the pro-longevity preservation of TCA cycle enzymatic activities and enhanced glycerol catabolism. These changes contribute positively to the phosphorylating respiration and to an increase in trehalose storage: both of which are longevity-promoting prerequisites.

Antioxidants doi: 10.3390/antiox14010079

Authors: Eleanor C. Ransdell-Green Janina Baranowska-Kortylewicz Dong Wang

Hydroxyl radicals (•OH), the most potent oxidants among reactive oxygen species (ROS), are a major contributor to oxidative damage of biomacromolecules, including DNA, lipids, and proteins. The overproduction of •OH is implicated in the pathogenesis of numerous diseases such as cancer, neurodegenerative disorders, and some cardiovascular pathologies. Given the localized nature of •OH-induced damage, detecting •OH, specifically near DNA and within organelles, is crucial for understanding their pathological roles. The major challenge of •OH detection results from their short half-life, high reactivity, and low concentrations within biological systems. As a result, there is a growing need for the development of highly sensitive and selective probes that can detect •OH in specific cellular regions. This review focuses on the advances in fluorescence probes designed to detect •OH near DNA and within cellular organelles and membranes. The key designs of the probes are highlighted, with emphasis on their strengths, applications, and limitations. Recommendations for future research directions are given to further enhance probe development and characterization.

Antioxidants doi: 10.3390/antiox14010078

Authors: Julia Tortajada-Pérez Andrea del Valle Carranza Cristina Trujillo-del Río Mar Collado-Pérez José María Millán Gema García-García Rafael Pascual Vázquez-Manrique

Lipid metabolism plays a critical role in maintaining cellular integrity, especially within the nervous system, where lipids support neuronal structure, function, and synaptic plasticity. However, this essential metabolic pathway is highly susceptible to oxidative stress, which can lead to lipid peroxidation, a damaging process induced by reactive oxygen species. Lipid peroxidation generates by-products that disrupt many cellular functions, with a strong impact on proteostasis. In this review, we explore the role of lipid oxidation in protein folding and its associated pathological implications, with a particular focus on findings in neurodegeneration from Caenorhabditis elegans studies, an animal model that remains underutilized. Additionally, we highlight the effectiveness of different methodologies applied in this nematode to deepen our understanding of this intricate process. In the nervous system of any animal, including mammals and invertebrates, lipid oxidation can disturb the delicate balance of cellular homeostasis, leading to oxidative stress, the build-up of toxic by-products, and protein misfolding, key factors in neurodegenerative diseases. This disruption contributes to the pathogenesis of neurodegenerative disorders such as Alzheimer’s, Parkinson’s, or Huntington’s disease. The findings from Caenorhabditis elegans studies offer valuable insights into these complex processes and highlight potential avenues for developing targeted therapies to mitigate neurodegenerative disease progression.

Antioxidants doi: 10.3390/antiox14010077

Authors: Smara Sigdel Shuzhen Chen Gideon Udoh Jinju Wang

Our group has recently demonstrated that exercise intervention affects the release and function of bone marrow endothelial progenitor cell-derived extracellular vesicles (EVs) in transgenic hypertensive mice. Whether such an exercise regimen can impact circulating EVs (cEVs) remains unknown. In this study, we investigated the influence of exercise on cEV level and function. Transgenic hypertensive mice (Alb1-Ren) underwent 8-week treadmill exercise (10 m/min for 1 h, 5 days per week). Age- and sex-matched sedentary Alb1-Ren mice served as controls. cEVs were isolated from the blood of exercised and sedentary mice and are denoted as ET-cEV and nET-cEV, respectively. cEVs were labeled to determine their uptake efficiency and pathways. The functions of cEVs were assessed in an Angiotensin II (Ang II) plus hypoxia-injured cerebral microvascular endothelial cell (mBMEC) injury model. Cellular migration ability and oxidative stress were evaluated. We found that treadmill exercise stimulated cEV release, and ET-cEVs were more prone to be internalized by mBMECs. The ET-cEV internalization was mediated by macropinocytosis and endocytosis pathways. Functional studies showed that ET-cEVs can improve the compromised migration capability of mBMECs challenged by Ang II plus hypoxia. Additionally, ET-cEV treatment upregulated the expression of p-Akt/Akt in mBMECs. Compared to nET-cEVs, ET-cEVs significantly reduced ROS overproduction in Ang II plus hypoxia-injured mBMECs, associated with decreased Nox2 expression. All these findings suggest that exercise-intervened cEVs can protect cerebral microvascular endothelial cells against hypertensive and hypoxic injury.

Antioxidants doi: 10.3390/antiox14010076

Authors: Fanni Ősz Aamir Nazir Krisztina Takács-Vellai Zsolt Farkas

Mutations in highly conserved genes encoding components of the electron transport chain (ETC) provide valuable insights into the mechanisms of oxidative stress and mitochondrial ROS (mtROS) in a wide range of diseases, including cancer, neurodegenerative disorders, and aging. This review explores the structure and function of the ETC in the context of its role in mtROS generation and regulation, emphasizing its dual roles in cellular damage and signaling. Using Caenorhabditis elegans as a model organism, we discuss how ETC mutations manifest as developmental abnormalities, lifespan alterations, and changes in mtROS levels. We highlight the utility of redox sensors in C. elegans for in vivo studies of reactive oxygen species, offering both quantitative and qualitative insights. Finally, we examine the potential of C. elegans as a platform for testing ETC-targeting drug candidates, including OXPHOS inhibitors, which represent promising avenues in cancer therapeutics. This review underscores the translational relevance of ETC research in C. elegans, bridging fundamental biology and therapeutic innovation.

Antioxidants doi: 10.3390/antiox14010075

Authors: Hamza Goujet Ismail Kaarouch Abderrahim Malki Modou Mamoune Mbaye Rosalie Cabry Noureddine Louanjli Taha Rhouda Moncef Benkhalifa

Sperm cryopreservation provides patients undergoing oncological, surgical, or infertility treatments the opportunity to conceive their own children, using assisted reproductive technologies. However, the freezing-thawing process can negatively influence both the quantity and the quality of spermatozoa, mainly due to an excessive production of reactive oxygen species and/or an impaired antioxidant defense system in sperm. Aromatic and medicinal plants synthesize essential oils with antioxidant proprieties as a part of their ecological adaptation to environmental stress, thanks to their rich bioactive phytochemical components. This study aimed to assess sperm progressive motility, viability, plasma membrane functionality, and lipid peroxidation levels of human cryopreserved normozoospermic (n = 51) and asthenozoospermic (n = 51) samples without or with the addition of Thymus satureoides (TSEO) (20 µg/mL), Artemisia vulgaris (AVEO) (48 µg/mL), and Rosmarinus officinalis (ROEO) (13 µg/mL) essential oils. Sperm parameters were significantly better preserved with ROEO in both normozoospermic (p < 0.05) and asthenozoospermic samples (p < 0.01). In contrast, TSEO had a negative impact for both groups (p < 0.05). Meanwhile, no significant effects were observed with AVEO. In summary, the study revealed that in vitro addition of essential oils as antioxidant agents during cryopreservation can be either beneficial, which helps preserve sperm parameters and fertilizing potential, or detrimental as spermicidal agents.

Antioxidants doi: 10.3390/antiox14010074

Authors: Muhammad Junaid Rao Bingsong Zheng

Plants have evolved complex mechanisms to cope with diverse abiotic stresses, with the phenylpropanoid pathway playing a central role in stress adaptation. This pathway produces an array of secondary metabolites, particularly polyphenols, which serve multiple functions in plant growth, development, regulating cellular processes, and stress responses. Recent advances in understanding the molecular mechanisms underlying phenylpropanoid metabolism have revealed complex regulatory networks involving MYB transcription factors as master regulators and their interactions with stress signaling pathways. This review summarizes our current understanding of polyphenol-mediated stress adaptations in plants, emphasizing the regulation and function of key phenylpropanoid pathway compounds. We discussed how various abiotic stresses, including heat and chilling stress, drought, salinity, light stress, UV radiation, nanoparticles stress, chemical stress, and heavy metal toxicity, modulate phenylpropanoid metabolism and trigger the accumulation of specific polyphenolic compounds. The antioxidant properties of these metabolites, including phenolic acids, flavonoids, anthocyanins, lignin, and polyphenols, and their roles in reactive oxygen species scavenging, neutralizing free radicals, membrane stabilization, and osmotic adjustment are discussed. Understanding these mechanisms and metabolic responses is crucial for developing stress-resilient crops and improving agricultural productivity under increasingly challenging environmental conditions. This review provides comprehensive insights into integrating phenylpropanoid metabolism with plant stress adaptation mechanisms, highlighting potential targets for enhancing crop stress tolerance through metabolic adjustment.

Antioxidants doi: 10.3390/antiox14010073

Authors: Alessandra Modesti Rosamaria Militello Alice Tanturli Alice Santi Massimo Gulisano Cristian Petri Luca Pengue Alessio Pellegrino Pietro Amedeo Modesti Simone Luti

Background: Male professional soccer players frequently compete in multiple matches weekly, and each match significantly impacts their homeostasis, health, and performance. This study evaluates players response at 48 h post-match by combining biological and GPS data. Investigating biochemical and performance metrics offers insights into the physical demands of high-intensity exercise, essential for optimizing performance, recovery, and overall athlete health. Methods: The study involved an Italian “Serie A” team, and we assessed players’ effort during a single match using GPS data and compared it to “Serie A” averages. Additionally, we evaluated oxidative stress and metabolism 48 h after the match. Results: At 48 h post-match, there were no signs of oxidative stress and changes in salivary IgA levels, but total antioxidant potential was significantly low. Moreover, increased plasma metabolites linked to energy production were also observed. Conclusions: The results indicate that 48 h after a match in “Serie A”, well-trained athletes showed no oxidative stress, to the detriment of the antioxidant potential, along with increased metabolites crucial for energy production. Combining GPS and metabolic analysis enhances player performance, informs tactical decisions, and supports team success, fostering data-driven approaches in soccer.

Antioxidants doi: 10.3390/antiox14010072

Authors: Alfredo Caturano Maria Rocco Giuseppina Tagliaferri Alessia Piacevole Davide Nilo Giovanni Di Lorenzo Ilaria Iadicicco Mariarosaria Donnarumma Raffaele Galiero Carlo Acierno Celestino Sardu Vincenzo Russo Erica Vetrano Caterina Conte Raffaele Marfella Luca Rinaldi Ferdinando Carlo Sasso

Type 2 diabetes mellitus (T2DM) is a chronic metabolic disorder that significantly increases the risk of cardiovascular disease, which is the leading cause of morbidity and mortality among diabetic patients. A central pathophysiological mechanism linking T2DM to cardiovascular complications is oxidative stress, defined as an imbalance between reactive oxygen species (ROS) production and the body’s antioxidant defenses. Hyperglycemia in T2DM promotes oxidative stress through various pathways, including the formation of advanced glycation end products, the activation of protein kinase C, mitochondrial dysfunction, and the polyol pathway. These processes enhance ROS generation, leading to endothelial dysfunction, vascular inflammation, and the exacerbation of cardiovascular damage. Additionally, oxidative stress disrupts nitric oxide signaling, impairing vasodilation and promoting vasoconstriction, which contributes to vascular complications. This review explores the molecular mechanisms by which oxidative stress contributes to the pathogenesis of cardiovascular disease in T2DM. It also examines the potential of lifestyle modifications, such as dietary changes and physical activity, in reducing oxidative stress and mitigating cardiovascular risks in this high-risk population. Understanding these mechanisms is critical for developing targeted therapeutic strategies to improve cardiovascular outcomes in diabetic patients.

Antioxidants doi: 10.3390/antiox14010071

Authors: Eun-Bin Kwon Buyun Kim Young-Eun Kim Sung-Joon Na Sang Mi Han Soon Ok Woo Hong Min Choi Siwon Moon Young Soo Kim Jang-Gi Choi

To combat influenza A virus (IAV) infection, it is vital to develop effective therapeutic strategies, including immunomodulators. In this study, we examined the antiviral effects of Hovenia dulcis Thunb. honey (HDH) against IAV using RAW 264.7 cells. HDH treatment significantly reduced IAV infection and viral protein expression. Moreover, it enhanced the production of interferon (IFN)-β, activated the innate immune response through the cyclic GMP–AMP synthase (cGAS)–stimulator of interferon genes (STING) pathway, and upregulated IFN signaling through signal transducer and activator of transcription (STAT)1/2 phosphorylation and interferon-stimulated gene (ISG) expression. In addition, HDH decreased IAV-induced intracellular and mitochondrial reactive oxygen species (ROS) production by upregulating the expression of antioxidant proteins, such as Sirt3 and SOD2. The results suggest that HDH is a potential therapeutic agent inhibiting viral replication and boosting host antiviral immunity.

Antioxidants doi: 10.3390/antiox14010070

Authors: Guangqi Guan Yaoxing Chen Yulan Dong

Oxidative stress (OS) refers to the production of a substantial amount of reactive oxygen species (ROS), leading to cellular and organ damage. This imbalance between oxidant and antioxidant activity contributes to various diseases, including cancer, cardiovascular disease, diabetes, and neurodegenerative conditions. The body’s antioxidant system, mediated by various signaling pathways, includes the AMPK-SIRT1-FOXO pathway. In oxidative stress conditions, AMPK, an energy sensor, activates SIRT1, which in turn stimulates the FOXO transcription factor. This cascade enhances mitochondrial function, reduces mitochondrial damage, and mitigates OS-induced cellular injury. This review provides a comprehensive analysis of the biological roles, regulatory mechanisms, and functions of the AMPK-SIRT1-FOXO pathway in diseases influenced by OS, offering new insights and methods for understanding OS pathogenesis and its therapeutic approaches.

Antioxidants doi: 10.3390/antiox14010069

Authors: Maciej Sylwester Bryś Krzysztof Olszewski Maciej Bartoń Aneta Strachecka

The increasing prevalence of monocultures has reduced floral diversity, diminishing pollen diet variety for bees. This study examines the impact of monofloral pollen diets (hazel, rapeseed, pine, buckwheat, Phacelia, goldenrod) on the antioxidant enzyme activities in the fat body from tergite 3, tergite 5, sternite, and hemolymph of honey bees. We show that pollen from plants such as rapeseed, Phacelia, buckwheat, and goldenrod (rich in phenolic compounds and flavonoids) increases the activities of SOD, CAT, GST, and GPx in the fat body and hemolymph compared to the control group. Although it is commonly known that a monodiet is one of the stress factors for bees, the increase in the activities of these enzymes in the hemolymph and fat body of workers fed with pollen candy compared to those fed only sugar candy has a positive (although inconclusive) effect. These activities in the hemolymph and fat body of bees fed with pollen from anemophilous plants are usually lower compared to those in bees fed with candy containing 10% pollen from rapeseed, Phacelia, buckwheat, or goldenrod. Further research is needed to fully understand the complex interactions among monofloral pollen diets, antioxidant enzyme activities, and the overall physiology of honey bees.

Antioxidants doi: 10.3390/antiox14010068

Authors: Lamprini Zissi Virginia D. Dimaki Vassiliki S. Birba Vassiliki C. Galani Vassiliki Magafa Sophia Hatziantoniou Fotini N. Lamari

This study investigated the potential of natural deep eutectic solvents (NADESs) for extracting bioactive compounds from the aerial parts of two mountain tea taxa, Sideritis clandestina ssp. peloponnesiaca (Boiss. & Heldr.) Baden and Sideritis raeseri Boiss. & Heldr. ssp. raeseri. Five NADEs, composed of betaine, glycerol, glucose, urea, citric acid, and sucrose, were evaluated for their extraction efficiency compared to conventional solvents (water and 70% ethanol). The total phenolic content (TPC) and antioxidant activity (FRAP and DPPH assays) were determined. Results showed that water was not a good extraction solvent. Despite the great solvent-dependent differences, most NADEs, particularly the betaine–glycerol–glucose mixture (BGG4), exhibited comparable or even superior extraction efficiency and antioxidant activity compared to 70% ethanol. The secondary metabolites in the BGG4 and 70% ethanol extracts were determined with HPLC-MS. The BGG4 extracts of both Sideritis taxa had a rich phenolic profile, with the major ingredients being chlorogenic acid, verbascoside, and non-, mono- and di-acetylated allosyl hypolaetin glycosides. Although distinct quantitative differences in their composition compared to the respective 70% ethanol extracts, and between them were noted, overall, the content of secondary metabolites in both S. raeseri extracts was lower than that of the S. clandestina extracts. These findings suggest that NADEs, particularly BGG4, are promising green solvents for extracting bioactive compounds from Sideritis taxa, paving the way for potential applications in the development of natural and sustainable cosmetic products.

Antioxidants doi: 10.3390/antiox14010067

Authors: Minhang Tu Gentan Cai Longfei Ma Leyan Yan Tian Wang Zhendan Shi Chao Wang Zhe Chen

Background: In vitro findings on the biological functions of Lycium barbarum flavonoids (LBFs) as feed additives are limited. This study aimed to explore the effects of different concentrations of LBFs on the growth performance, immune function, intestinal barrier, and antioxidant capacity of meat ducks. A total of 240 one-day-old male meat ducks were randomly allocated to four groups, each receiving a basal diet supplemented with 0 (control), 250, 500, or 1000 mg/kg of LBFs for 42 d. Results: The results showed that dietary supplementation with 500 mg/kg of LBFs resulted in a significant increase in average daily feed intake, body weight, average daily gain, and feed conversion ratio. Dietary supplementation with 500 or 1000 mg/kg of LBFs resulted in significant decreases in serum levels of D-lactic acid and lipopolysaccharide. Dietary supplementation with 500 mg/kg LBFs significantly decreased diamine oxidase activity and enhanced the activities of catalase, total antioxidant capacity, and glutathione peroxidase in the jejunal mucosa, as well as the activity of total superoxide dismutase and the content of glutathione in the ileal mucosa, while significantly lowering the content of malondialdehyde in the ileal mucosa. Dietary supplementation with 500 mg/kg LBFs significantly up-regulated the mRNA expression of genes associated with intestinal barrier function and antioxidant capacity in the jejunal and ileal mucosa, as well as the protein expression of these antioxidant genes, and led to a significant reduction in the mRNA expression of pro-apoptotic and inflammatory-related genes. Conclusions: The addition of LBFs to the diet improved the growth performance, intestinal barrier function, immune response, and antioxidant capacity of the ducks, which may be closely associated with the activation of the Nrf2 signaling pathway and the inhibition of the NF-κB signaling pathway. The optimal dietary inclusion level of LBFs in ducks was 500 mg/kg.

Antioxidants doi: 10.3390/antiox14010066

Authors: Dario Troise Costanza Allegra Luciana Antonia Cirolla Silvia Mercuri Barbara Infante Giuseppe Castellano Giovanni Stallone

The complement system plays a crucial role in regulating the inflammatory responses in kidney transplantation, potentially contributing to early decline in kidney function. Ischemia–reperfusion injury (IRI) is among the factors affecting graft outcomes and a primary contributor to delayed graft function. Complement activation, particularly the alternative pathway, participates in the pathogenesis of IRI, involving all kidney compartments. In particular, tubular epithelial cells often acquire a dysfunctional phenotype that can exacerbate complement activation and kidney damage. Currently, complement-modulating drugs are under investigation for the treatment of kidney diseases. Many of these drugs have shown potential therapeutic benefits, but no effective clinical treatments for renal IRI have been identified yet. In this review, we will explore drugs that target complement factors, complement receptors, and regulatory proteins, aiming to highlight their potential value in improving the management of renal IRI.

Antioxidants doi: 10.3390/antiox14010065

Authors: Pilar Dominguez-Verano Nadia Jacobo-Herrera Andrés Castell-Rodríguez Octavio Canales-Alvarez Maria Margarita Canales-Martinez Marco Aurelio Rodriguez-Monroy

Propolis is a resinous substance produced by bees that has several biomedical properties that could contribute to the repair process of the gastric mucosa, such as antioxidant, anti-inflammatory, healing, and gastroprotective properties. Thus, this study aimed to determine the chemical composition of Mexicali propolis, its antioxidant capacity, and its effect on gastric repair. Three polarity-directed extracts were obtained: the ethanolic extract, the ethyl acetate extract, and the hexane extract. The antioxidant activity, total phenolic content (TPC), and flavone/flavonol content were determined for each extract. The chemical composition was analysed using HPLC—TOF—MS (High—Performance Liquid Chromatography—Time—Of—Flight Mass Spectrometry) and GC—MS (Gas Chromatography–Mass Spectrometry), and a total of 52 compounds were identified. The results revealed that the ethanolic extract had the greatest effect on free radical scavenging and the content of bioactive compounds. On the basis of these results, the effect of the Mexicali ethanolic extract of propolis (MeEEP) on gastric repair was subsequently evaluated. Prior to the evaluation, MeEEP was found to exhibit low oral toxicity, as determined under the Organisation for Economic Co-operation and Development (OECD) 425 guidelines. Gastric injury was induced in male C57BL/6 mice by intragastric administration of indomethacin (10 mg/kg). MeEEP (300 mg/kg) was administered 6 h after the induction of injury using indomethacin and daily thereafter. The mice were sacrificed at 12, 24, and 48 h to assess the effect. As a result, MeEEP enhanced the repair of the gastric lesion by decreasing the percentage of the bleeding area and attenuating the severity of histological damage, as demonstrated by H&E staining. This effect was associated with a reduction in MPO enzyme activity and in the levels of the proinflammatory cytokines TNF-α, IL-1β, and IL-6, maintaining controlled inflammation in gastric tissue. Furthermore, the administration of the extract increased SOD enzymatic activity and GSH levels, reducing the degree of oxidative damage in the gastric tissue, as demonstrated by low MDA levels. Finally, after evaluating the effect on apoptosis via immunohistochemistry, MeEEP was shown to reduce the expression of the proapoptotic marker Bax and increase the expression of the antiapoptotic marker Bcl-2. In conclusion, these findings suggest that MeEEP may enhance gastric repair through a cytoprotective mechanism by controlling inflammation exacerbation, reducing oxidative stress, and regulating apoptosis. These mechanisms are primarily attributed to the presence of pinocembrin, tectochrysin, chrysin, apigenin, naringenin, acacetin, genistein, and kaempferol. It is important to highlight that this study provides a preliminary exploration of the reparative effect of Mexican propolis, describing the potential mechanisms of action of the compounds present in Mexicali propolis.

Antioxidants doi: 10.3390/antiox14010064

Authors: Ivana Tlak Gajger Josipa Vlainić

Antioxidants have gained significant importance in modern nutrition [...]

Antioxidants doi: 10.3390/antiox14010063

Authors: Lei Jiang Minggang Xiao Rongfeng Huang Juan Wang

Salinity affects crop growth and productivity, and this stress can be increased along with drought or high temperature stresses and poor irrigation management. Cultivation of salt-tolerant crops plays a critical role in enhancing crop yield under salt stress. In the past few decades, the mechanisms of plant adaptation to salt stress have been described, especially relying on ionic homeostasis, reactive oxygen species (ROS) scavenging, and phytohormone signaling. The studies of these molecular mechanisms have provided a basis for breeding new salt-tolerant crop germplasm and have facilitated the entry into the era of molecular breeding of salt-tolerant crops. In this review, we outline the recent progress in the molecular regulations underlying crop salt tolerance, focusing on the double-edged sword effect of ROS, the regulatory role of phytohormones, and the trade-off effects of ROS and phytohormones between crop yield and salt tolerance. A future challenge is to identify superior alleles of key salt-tolerant genes that will accelerate the breeding of high-yield and salt-tolerant varieties.

Antioxidants doi: 10.3390/antiox14010062

Authors: Sho Nishida Tamaki Ishima Daiki Iwami Ryozo Nagai Kenichi Aizawa

Tacrolimus (TAC)-induced chronic nephrotoxicity (TAC nephrotoxicity) is a serious issue for long-term graft survival in kidney transplantation. However, the pathophysiology of TAC nephrotoxicity remains unclear. In this study, we analyzed whole blood samples from mice that developed TAC nephrotoxicity in order to discover its mechanism. Mice were divided into a TAC group and a control group (n = 5 per group). The TAC group received TAC subcutaneously (1 mg/kg/day for 28 days), while the control group received normal saline instead. After the administration period, whole blood was collected and metabolomic analysis was performed, revealing significant changes in 56 metabolites. The major metabolic changes were related to uremic toxins, vascular damage, and NAD+. NAD+ levels were significantly lower in the TAC group, and ADP-ribose, nicotinamide, and nicotinamide N-oxide, which are degradation products of NAD+, were significantly higher, suggesting impairment of the NAD+ salvage pathway. NAD+ deficiency suggests cellular aging and mitochondrial dysfunction, which may induce vascular damage and chronic kidney disease. Our study demonstrated a correlation between low NAD+ levels and the pathophysiology of TAC nephrotoxicity.

Antioxidants doi: 10.3390/antiox14010061

Authors: Mina Park Sunghye Cho Dongtak Jeong

Cardiac ischemia–reperfusion injury (IRI) occurs when blood flow is restored to the myocardium after a period of ischemia, leading to oxidative stress and subsequent myocardial cell damage, primarily due to the accumulation of reactive oxygen species (ROS). In our previous research, we identified that miR-25 is significantly overexpressed in pressure overload-induced heart failure, and its inhibition improves cardiac function by restoring the expression of SERCA2a, a key protein involved in calcium regulation. In this study, we aimed to investigate the role of miR-25 in the context of ischemia–reperfusion injury. We found that miR-25 was markedly upregulated under hypoxic conditions in both in vitro and in vivo models. Through in silico analysis, we identified Sestrin3 (SESN3), an antioxidant protein known for its protective effects against oxidative stress, as a novel target of miR-25. Based on these findings, we hypothesized that inhibiting miR-25 would restore Sestrin3 expression, thereby reducing ROS-induced myocardial cell damage and improving cardiac function. To test this hypothesis, we employed two model systems: a hypoxia/reoxygenation (H/R) stress model using H9c2 myoblasts and a surgically induced ischemia–reperfusion injury mouse model. Our results demonstrated that the use of miR-25 inhibitors significantly improved cardiac function and reduced myocardial damage in both models through the restoration of SESN3 expression. In conclusion, our findings suggest that targeting miR-25 may serve as a novel therapeutic modality to alleviate oxidative damage in the heart.

Antioxidants doi: 10.3390/antiox14010060

Authors: Abdullah S. Seddiek Kaiwen Chen Fanlin Zhou Muhindo Mwizerwa Esther Abdelaziz Elbarbary Hazem Golshany Angelo Uriho Li Liang

Whey protein isolate (WPI) has functional properties such as gelation and emulsification. Emulsion gels combine the benefits of both emulsions and hydrogels. In this study, WPI hydrogels and emulsion gels were developed with goji oil (GO) as the oil phase by the inclusion of blueberry extract (BE) in the protein matrix. Heat-denatured WPI (hWPI) particles and emulsions were characterized in terms of size distribution, ζ-potential, interfacial protein, and anthocyanin partition. The inclusion of anthocyanins-rich blueberry extract led to the aggregation of hWPI particles, but it also increased the interfacial protein of 10% goji oil emulsions to 20% and decreased their size distribution to 120 and 325 nm. WPI hydrogels and emulsion gels were analyzed in terms of their water-holding capacity, which decreased from 98% to 82% with the addition of blueberry extract and goji oil. Syneresis, rheological, and morphological characteristics were also analyzed. The gelation time of hWPI particles and emulsions was shortened from 24 h to 12 h when incorporating blueberry extract to form a dense network. The network was the most homogeneous and densest in the presence of 3% blueberry extract and 5% goji oil. The co-inclusion of blueberry extract and goji oil increased the syneresis during the freeze–thaw cycles, with the values rising from 13% to 36% for 5% BE hydrogel and BE-containing emulsion gels after the first cycle. All WPI hydrogels and emulsion gels exhibit predominantly elastic behavior. Moreover, anthocyanin release, antioxidant activity, and the fatty acid composition profile were also analyzed during in vitro digestion. Soluble and free anthocyanins in the digested medium were reduced with the goji oil content but increased with the blueberry extract content. The stability of polyunsaturated fatty acids in the digested medium was improved by the addition of blueberry extract. The antioxidant activity of the digested medium increased with the content of blueberry extract but decreased with the content of goji oil. The ABTS∙+ scavenging capacities decreased from 63% to 49% by increasing the content of GO from 0% to 10% and they increased from 48% to 57% for 5% BE and 10% GO emulsion gels as the BE content increased from 0% to 5% after 6 h of digestion. The data gathered should provide valuable insights for future efforts to co-encapsulate hydrophilic and hydrophobic agents, thereby enhancing their stability, bioavailability, and functional properties for potential applications in food industries.

Antioxidants doi: 10.3390/antiox14010059

Authors: Marin Kuntić Ivana Kuntić Jiayin Zheng Leonardo Nardi Matthias Oelze Arijan Valar Dominika Mihaliková Lea Strohm Henning Ubbens Qi Tang Liyu Zhang Guilherme Horta Paul Stamm Omar Hahad Dilja Krueger-Burg Huige Li Sebastian Steven Adrian Gericke Michael J. Schmeisser Thomas Münzel Andreas Daiber

Noise pollution is a known health risk factor and evidence for cardiovascular diseases associated with traffic noise is growing. At least 20% of the European Union’s population lives in noise-polluted areas with exposure levels exceeding the recommended limits of the World Health Organization, which is considered unhealthy by the European Environment Agency. This results in the annual loss of 1.6 million healthy life years. Here, we investigated the protective effects of cardiovascular drug interventions against aircraft noise-mediated cardiovascular complications such as elevated oxidative stress or endothelial dysfunction. Using our established mouse exposure model, we applied mean sound pressure levels of 72 dB(A) for 4 d. C57BL/6 mice were treated with the beta-blocker propranolol (15 mg/kg/d s.c. for 5 d) or the alpha-blocker phenoxybenzamine (1.5 mg/kg/d s.c. for 5 d) and noise-exposed for the last 4 d of the drug administration. Short-term noise exposure caused hypertension (measured by tail-cuff blood pressure monitoring) and impaired endothelial function (measured by isometric tension recording in the aorta and video microscopy in cerebral arterioles in response to acetylcholine). Noise also increased markers of oxidative stress and inflammation. Treatment of mice with propranolol and phenoxybenzamine prevented endothelial and microvascular dysfunction, which was supported by a decrease in markers of inflammation and oxidative stress in heart tissue and the brain. Amelioration of noise-induced hypertension (systolic blood pressure) was not observed, whereas pulse pressure was lowered by trend. This study provides a novel perspective mitigating the adverse effects of noise pollution, especially in vulnerable groups with medication, a rationale for further pharmacological human studies.

Antioxidants doi: 10.3390/antiox14010058

Authors: Xin Xu Hongwei Zhang Kun Meng Hongying Cai Weiwei Liu Liye Song Zihan Zhang Qijun Zhu Xiling Han Yunsheng Han Peilong Yang

Limosilactobacillus reuteri, a recognized probiotic, improves intestinal health in animals, but the mechanism remains unclear. This study investigates the mechanisms by which L. reuteri ZY15, isolated from healthy pig feces, mitigates intestinal barrier damage and inflammation caused by oxidative stress in Enterotoxigenic Escherichia coli (ETEC) K88-challenged mice. The results indicated that L. reuteri ZY15 increased antioxidant capacity by reducing serum reactive oxygen species (ROS) and superoxide dismutase (SOD) levels. L. reuteri ZY15 enhanced the intestinal barrier by upregulating mucin 1, mucin 2, occludin, zonula occludens-1 (ZO-1), and claudin-1 expressions in protein and mRNA levels. It significantly alleviated intestinal inflammation by reducing the proinflammatory cytokines interleukin-1β (IL-1β), interferon-γ (IFN-γ), tumor necrosis factor-α (TNF-α), and interleukin-17 (IL-17) mRNA and protein levels. Notably, L. reuteri ZY15 suppressed intestinal inflammation by inhibiting AKT/mTOR/HIF-1α/RORγt/IL-17 pathway activation. Additionally, it significantly altered the structure of gut microorganisms by enriching Akkermansia and Clostridia_UCG.014, and thereby re-establishing colonization resistance and alleviating ETEC K88-induced intestinal barrier damage and inflammation in mice. Taken together, our findings reveal the protective mechanism of L. reuteri ZY15 in mice challenged with ETEC K88 by regulating AKT/mTOR/HIF-1α/RORγt/IL-17 signaling and microbial imbalance. Leveraging these properties, live L. reuteri ZY15 offers a promising alternative treatment for Escherichia coli-induced diarrhea in weaned piglets.

Antioxidants doi: 10.3390/antiox14010057

Authors: Ying Wang Lanjie Li Danni Chen Jiaheng Shan Meijuan Yi Hiroyuki Osada Minoru Yoshida Lan Xiang Jianhua Qi

Geniposidic 4-isoamyl ester (GENI) with anti-aging effects is a new iridoid glycoside derivative from Gardenia jasminoides Ellis found in our previous study. In this study, to indicate whether this compound has anti-Alzheimer’s disease (AD) effect, the galactose-induced AD mice and naturally aging mice with AD were used to do drug efficacy evaluation. Furthermore, the Western blot, small interfering RNA (siRNA), drug affinity responsive target stability (DARTS), cellular thermal shift assay (CESTA), liquid chromatography-tandem mass spectrometry (LC/MS-MS), adenosine 5′-monophosphate-activated protein kinase (AMPK) mutants and surface plasmon resonance (SPR) analysis were utilized to clarify the mechanism of action and identify target protein of this molecule. GENI exerts anti-AD efficacy in galactose-induced AD mice and naturally aging mice with AD through neuroprotection and modification of autophagy and neuron inflammation. Moreover, AMPK as the target protein of GENI to produce an anti-AD effect is identified and the ASP148, ASP157, and ASP166 of the AMPK α subunit and lysine (LYS)148, aspartic acid (ASP)156, LYS309, and ASP316 in the AMPK γ subunit as binding sites are confirmed. Meanwhile, the AMPK/unc-51-like autophagy-activating kinase 1 (ULK1)/microtubule-associated protein 1 light chain 3 beta (LC3B) and AMPK/mammalian target of rapamycin (mTOR) signaling pathways involved in anti-AD effects of GENI. The findings provide a new perspective on treating neurodegenerative diseases by activating AMPK for the energy metabolism disorder.

Antioxidants doi: 10.3390/antiox14010056

Authors: Fabrizio Termite Sebastiano Archilei Francesca D’Ambrosio Lucrezia Petrucci Nicholas Viceconti Roberta Iaccarino Antonio Liguori Antonio Gasbarrini Luca Miele

Metabolic dysfunction-associated steatotic liver disease (MASLD) is a prevalent chronic liver condition marked by excessive lipid accumulation in hepatic tissue. This disorder can lead to a range of pathological outcomes, including metabolic dysfunction-associated steatohepatitis (MASH) and cirrhosis. Despite extensive research, the molecular mechanisms driving MASLD initiation and progression remain incompletely understood. Oxidative stress and lipid peroxidation are pivotal in the “multiple parallel hit model”, contributing to hepatic cell death and tissue damage. Gut microbiota plays a substantial role in modulating hepatic oxidative stress through multiple pathways: impairing the intestinal barrier, which results in bacterial translocation and chronic hepatic inflammation; modifying bile acid structure, which impacts signaling cascades involved in lipidic metabolism; influencing hepatocytes’ ferroptosis, a form of programmed cell death; regulating trimethylamine N-oxide (TMAO) metabolism; and activating platelet function, both recently identified as pathogenetic factors in MASH progression. Moreover, various exogenous factors impact gut microbiota and its involvement in MASLD-related oxidative stress, such as air pollution, physical activity, cigarette smoke, alcohol, and dietary patterns. This manuscript aims to provide a state-of-the-art overview focused on the intricate interplay between gut microbiota, lipid peroxidation, and MASLD pathogenesis, offering insights into potential strategies to prevent disease progression and its associated complications.

Antioxidants doi: 10.3390/antiox14010055

Authors: Kazushi Watanabe Hiroyuki Kinoshita Tomohito Okamoto Kazumasa Sugiura Shingo Kawashima Tetsuro Kimura

Albumin, the most abundant protein, contributes significantly to various physiological processes, indicating its multifunctional properties. It has drawn the attention of scientists and physicians because of its primary role in maintaining osmotic pressure and involvement in transporting numerous small molecules, including hormones, fatty acids, and drugs. A growing body of evidence has recently illustrated an additional aspect of albumin’s antioxidant properties. Therefore, based on recent research findings, this review article delves into the molecular and biochemical aspects of albumin’s antioxidative capabilities. We highlight the multifaceted significance of proteins in oxidative stress and their relation to pathologies in obstetrics and gynecology. In particular, we focused on preeclampsia, in which oxidative stress is closely involved in the pathogenesis, and renal dysfunction leads to increased albumin excretion into the urine, resulting in hypoalbuminemia. In addition, we discussed the role of albumin in preeclampsia pathogenesis, diagnosis, and patient prognosis. Understanding the antioxidant properties of albumin opens new avenues for therapeutic intervention and sheds light on novel strategies for combating preeclampsia associated with oxidative damage. In this study, we employed the PubMed database to search for articles that assessed the antioxidant properties of albumin, with a specific focus on obstetric diseases, particularly preeclampsia. The last update of the search was conducted in January 2024.

Antioxidants doi: 10.3390/antiox14010054

Authors: Alessia Silla Angela Punzo Francesca Bonvicini Matteo Perillo Marco Malaguti Antonello Lorenzini Ismaela Foltran Dario Mercatante Mara Mandrioli Maria Teresa Rodriguez-Estrada Silvana Hrelia Cristiana Caliceti

Agricultural food waste and by-products could provide high-value compounds that positively affect human and environmental health, thus representing promising ingredients for cosmeceutical products. This study explores the biological activities of tomato skin (HP) and pomegranate peel (PPE) extracts on oral mucosa to evaluate their possible use in mouthwashes. The biological activities of the extracts and the mouthwash (MW) containing them were evaluated in Human Primary Gingival Epithelial cells (HGECs). The antioxidant and anti-inflammatory activities were analyzed in HGECs injured with lipopolysaccharides. After 24 h of treatment with PPE, HP, and MW, significant antioxidant activity and an increased Superoxide Dismutase 1 expression (p < 0.01) were observed. Additionally, the extracts significantly reduced the expression of tumor necrosis factor α (p < 0.05) and Monocyte Chemoattractant Protein 1 (p < 0.001), suggesting an anti-inflammatory role. Lastly, the antibacterial activity was assessed against Streptococcus mutans and Streptococcus sanguinis by the broth microdilution method and agar cup diffusion test for the extracts and the mouthwash, respectively, demonstrating strong effectiveness against both oral streptococcus species. Results demonstrate the potential of HP and PPE in reducing oxidative stress, inflammation, and bacterial proliferation within oral mucosa, highlighting food waste up-cycling as a resource for human health.

Antioxidants doi: 10.3390/antiox14010053

Authors: Angelo Santoro Antonio Ricci Manuela Rodriquez Michela Buonocore Anna Maria D’Ursi

Investigating amyloid–β (Aβ) peptides in solution is essential during the initial stages of developing lead compounds that can influence Aβ fibrillation while the peptide is still in a soluble state. The tendency of the Aβ(1–42) peptide to misfold in solution, correlated to the aetiology of Alzheimer’s disease (AD), is one of the main hindrances to characterising its aggregation kinetics in a cell-mimetic environment. Moreover, the Aβ(1–42) aggregation triggers the unfolded protein response (UPR) in the endoplasmic reticulum (ER), leading to cellular dysfunction and multiple cell death modalities, exacerbated by reactive oxygen species (ROS), which damage cellular components and trigger inflammation. Antioxidants like curcumin, a derivative of Curcuma longa, help mitigate ER stress by scavenging ROS and enhancing antioxidant enzymes. Furthermore, evidence in the literature highlights the effect of curcumin on the secondary structure of Aβ(1–42). This explorative study investigates the Aβ(1–42) peptide conformational behaviour in the presence of curcumin and six derivatives using circular dichroism (CD) to explore their interactions with lipid bilayers, potentially preventing aggregate formation. The results suggest that the synthetic tetrahydrocurcumin (THC) derivative interacts with the amyloid peptide in all the systems presented, while cyclocurcumin (CYC) and bisdemethoxycurcumin (BMDC) only interact when the peptide is in a less stable conformation. Molecular dynamics simulations helped visualise the curcuminoids’ effect in an aqueous system and hypothesise the importance of the peptide surface exposition to the solvent, differently modulated by the curcumin derivatives.

Antioxidants doi: 10.3390/antiox14010052

Authors: Dongyu Huang Jiaze Gu Hualiang Liang Mingchun Ren Chunyu Xue

A seven-week trial was designed to evaluate the effects of dietary seaweed polysaccharide (SP) supplementation on the growth performance and physiological health of largemouth bass. The results reveal that the 0.05SP group showed the best growth performance. The mRNA expression levels of tor, 4ebp1, and igf1 genes were remarkably down-regulated in the 0.15SP and 0.2SP groups compared to the control group. The CAT activities were significantly increased in the 0.05SP and 0.1SP groups, and the GSH-Px activity was increased in the 0.15SP group. The expression of the immune response-related gene nfκb was significantly down-regulated in the 0.1SP group, and those of tnfα and il-8 were at the maximum in the control group. Moreover, the expression of il-10 in the 0.15SP and 0.2SP groups was significantly down-regulated. Furthermore, endoplasmic reticulum stress (ERS)-related expression of atf6 was the highest in the control group. Furthermore, the chopα and bax expression levels in the 0.15SP and 0.2SP groups were significantly down-regulated compared with other groups. In addition, the highest expression level of bcl-xl was observed in the 0.15SP group. Finally, the quadratic regression analysis of antioxidant, immune, and ERS core parameters (CAT, nf-κb, and bcl-xl) determined 0.06–0.11% to be the optimal SP supplemental level in largemouth bass diets.

Antioxidants doi: 10.3390/antiox14010051

Authors: Fan Chen Yining Zhao Yanfa Dai Ning Sun Xuezheng Gao Jiajun Yin Zhenhe Zhou Ke-jia Wu

Ulcerative colitis (UC) is a chronic immune disease that is difficult to cure. We recently found that chick early amniotic fluid (ceAF) has notable anti-inflammatory and antioxidative properties, through its active components. This study demonstrates the potential of ceAF as a protective agent against UC. UPLC-MS mass spectrometry identified key components of ceAF, including various fatty acids and nucleosides. In vitro, ceAF improved viability in DSS-induced Caco-2 cells, reduced pro-inflammatory cytokines IL-1β and TNF-α, and increased the anti-inflammatory cytokine IL-10. It also upregulated the tight junction proteins ZO-1 and occludin. In DSS-induced UC mice, ceAF treatment alleviated weight loss, colon shortening, and disease activity, while improving histopathology, crypt depth, and colonic fibrosis. Mechanistically, ceAF’s anti-inflammatory effects are mediated by inhibiting the overactivation of TCR signaling through the LCK/ZAP70/LAT pathway. Our findings suggest that ceAF could be a valuable nutritional intervention for UC, potentially enhancing existing functional foods aimed at managing this condition.

Antioxidants doi: 10.3390/antiox14010050

Authors: Jirawadee Kaewda Surintorn Boonanuntanasarn Papungkorn Sangsawad Pimpisut Manassila Chatsirin Nakharuthai

Due to its lack of the L-gulonolactone oxidase (GULO) enzyme, Nile tilapia is unable to synthesize vitamin C; thus, it requires an adequate level of exogenous vitamin C in its diet. To enhance antioxidant properties and vitamin C-related effects, we employed recombinant technology to integrate the GULO-encoding gene into the Bacillus subtilis chromosome. In this study, fish were divided into four groups: those fed with a basal diet (CON), a basal diet + vitamin C (VC), a basal diet + wild-type B. subtilis (BS), and a basal diet + recombinant B. subtilis (BS+GULO). After 90 days of the feeding trial, the BS+GULO groups showed the highest improvements in final weight, weight gain, specific growth rate, average daily gain, and relative growth rate. The VC, BS, and BS+GULO groups exhibited increased total immunoglobulin and lysozyme activity; however, only the VC and BS+GULO groups showed elevated alternative complement 50 levels, phagocytic activity and improved antioxidant parameters compared to the control. HPLC and qRT-PCR analyses revealed elevated serum vitamin C and intestinal GULO mRNA levels in the BS+GULO group. A challenge test showed increased pro-inflammatory gene expression and immune response against S. agalactiae in the BS+GULO group, indicating improved antagonistic activity over wild-type B. subtilis.

Antioxidants doi: 10.3390/antiox14010049

Authors: Eunsun Lee Dongki Yang Jeong Hee Hong

The relationship between oxidative stress and cancer has been extensively studied and highlighted, along with its role in various aspects of angiogenesis. The modulation of oxidative levels and the adaptive mechanisms of oxidative stress in cancer systems are attractive research themes for developing anti-cancer strategies. Reactive oxygen species (ROS) are involved in various pathophysiological processes and play crucial roles in DNA damage and angiogenesis. Although cancer cells have developed various adaptive defense mechanisms against oxidative stress, excessive ROS production has been proposed as an anti-cancer strategy to induce cellular apoptosis. In particular, natural-source-based antioxidants have been identified as effective against cancers, and various delivery platforms have been developed to enhance their efficacy. In this review, we highlighted the anti-cancer components (plumbagin, quercetin, resveratrol, curcumin, xanthatin, carvacrol, telmisartan, and sulforaphane) that modulate ROS levels and the recent targeting platforms used to increase the application of anti-cancer drugs and the developed delivery platforms with diverse mechanisms of action. Further, we summarized the actual doses used and the effects of these drug candidates in various cancer systems. Overall, this review provides beneficial research themes for expanding cancer-targeting fields and addressing limited applications in diverse cancer types.

Antioxidants doi: 10.3390/antiox14010048

Authors: Francesca Gorini Alessandro Tonacci

Congenital heart disease (CHD) represents the major cause of infant mortality related to congenital anomalies globally. The etiology of CHD is mostly multifactorial, with environmental determinants, including maternal exposure to ambient air pollutants, assumed to contribute to CHD development. While particulate matter (PM) is responsible for millions of premature deaths every year, overall ambient air pollutants (PM, nitrogen and sulfur dioxide, ozone, and carbon monoxide) are known to increase the risk of adverse pregnancy outcomes. In this literature review, we provide an overview regarding the updated evidence related to the association between maternal exposure to outdoor air pollutants and CHD occurrence, also exploring the underlying biological mechanisms from human and experimental studies. With the exception of PM, for which there is currently moderate evidence of its positive association with overall CHD risk following exposure during the periconception and throughout pregnancy, and for ozone which shows a signal of association with increased risk of pooled CHD and certain CHD subtypes in the periconceptional period, for the other pollutants, the data are inconsistent, and no conclusion can be drawn about their role in CHD onset. Future epidemiological cohort studies in countries with different degree of air pollution and experimental research on animal models are warranted to gain a comprehensive picture of the possible involvement of ambient air pollutants in CHD etiopathogenesis. While on the one hand this information could also be useful for timely intervention to reduce the risk of CHD, on the other hand, it is mandatory to scale up the use of technologies for pollutant monitoring, as well as the use of Artificial Intelligence for data analysis to identify the non-linear relationships that will eventually exist between environmental and clinical variables.

Antioxidants doi: 10.3390/antiox14010047

Authors: Mario de la Fuente-Muñoz Marta Román-Carmena Sara Amor María C. Iglesias-de la Cruz Patricia Martorell Sonia Guilera-Bermell Reme García Bou Antonio M. Inarejos-García Ángel L. García-Villalón Miriam Granado

Arterial hypertension has a high prevalence in the population and is considered both a cardiovascular disease and an important risk factor for the development of other cardiovascular diseases. Tea consumption shows antihypertensive effects due to its composition in terms of bioactive substances such as flavan-3-ols and xanthines. The aim of this study was to assess the possible beneficial effects of two tea extracts, one of white tea (ADM® White Tea; WTE) and another one composed of a mixture of black tea and green tea (ADM® Tea Complex; CTE), on the cardiovascular alterations induced by angiotensin II (AngII) infusion in mice. For this purpose, four groups of C57BL/6J male mice were used: (1) mice fed on a standard diet for 8 weeks and infused with saline for the last 4 weeks (controls); (2) mice fed on a standard diet for 8 weeks and infused with AngII for the last 4 weeks (AngII); (3) mice fed on a standard diet supplemented with 1.6% WTE and infused with AngII for the last 4 weeks (AngII + WTE); (4) mice fed on a standard diet supplemented with 1.6% TC and infused with AngII for the last 4 weeks (AngII + CTE). Both tea extracts exerted anti-inflammatory and antioxidant effects in arterial tissue and reduced AngII-induced endothelial dysfunction in aorta segments. Moreover, supplementation with WTE or CTE reduced the Ang-II-induced overexpression of AT1R and increased AngII-induced downregulation of AT2R in arterial tissue. However, only supplementation with CTE significantly increased the circulating levels of angiotensin 1-7 and reduced systolic blood pressure. In the heart, supplementation with both tea extracts attenuated AngII-induced cardiac hypertrophy and reduced ischemia-reperfusion-induced oxidative stress and apoptosis in myocardial tissue. In conclusion, supplementation with WTE or CTE attenuates AngII-induced cardiovascular damage through their anti-inflammatory, antioxidant, and antiapoptotic effects. In addition, supplementation with CTE also exerts antihypertensive effects, and so it may constitute an avenue through which to support cardiovascular health.

Antioxidants doi: 10.3390/antiox14010046

Authors: Ignacija Vlašić Antonio Krstačić-Galić Anđela Horvat Nada Oršolić Anja Sadžak Lucija Mandić Suzana Šegota Maja Jazvinšćak Jembrek

Although commonly appreciated for their anti-oxidative and neuroprotective properties, flavonoids can also exhibit pro-oxidative activity, potentially reducing cell survival, particularly in the presence of metal ions. Disrupted copper homeostasis is a known contributor to neuronal dysfunction through oxidative stress induction. This study investigated the effects of myricitrin (1–20 μg/mL) on copper-induced toxicity (0.5 mM CuSO4) in the neuroblastoma SH-SY5Y cell line. At non-toxic concentrations, myricitrin exacerbated copper’s toxic effects. The myricitrin-induced decrease in survival was accompanied with increased reactive oxygen species (ROS) production, reduced superoxide dismutase activity, and a lower GSH/GSSG ratio. In combination with copper, myricitrin also activated caspase-3/7, promoted nuclear chromatin changes, and compromised membrane integrity. At the protein level, myricitrin upregulated p53 and PUMA expression. The toxic effects of myricitrin were alleviated by the p38 inhibitor SB203580, the intracellular calcium chelator BAPTA-AM, and the NMDA receptor blocker MK-801, highlighting the significant role of the ROS/p53/p38 axis in cell death and the critical involvement of calcium ions in apoptosis induction. The atomic force microscopy was used to assess the surface morphology and nanomechanical properties of SH-SY5Y cells, revealing changes following myricitrin treatment. This research highlights the toxic potential of myricitrin and emphasizes the need for caution when considering flavonoid supplementation in conditions with elevated copper levels.

Antioxidants doi: 10.3390/antiox14010044

Authors: Hao Wu Baochun Qin Guang Yang Pengyun Ji Yu Gao Lu Zhang Bingyuan Wang Guoshi Liu

As the global climate changes, high temperatures will cause heat stress, which significantly affects the productive efficiency of livestock. Currently, there is a lack of efficient methods to use in targeting this issue. In this study, we report that melatonin supplementation may represent an alternative method to reduce the negative impact of heat stress on livestock, particularly in Hainan black goats. Our results show that melatonin treatment increased the average daily gain of Hainan black goats that were exposed to constantly high temperatures for two months compared to controls. Our mechanistic exploration revealed that melatonin treatment not only reduced the oxidative stress and inflammatory reaction caused by heat stress but also improved goats’ metabolic capacity, promoting their growth and development. More importantly, for the first time, we observed that melatonin treatment modified the abundance of the intestinal microflora, altering the metabolism of the goats, which further improved their tolerance to constant heat stress.

Antioxidants doi: 10.3390/antiox14010045

Authors: Min Jiang Xuekai Zhang Xiao Jin Binlin Shi Yuanqing Xu Zheqi Wang

Artemisia ordosica Krasch, a traditional Chinese herbal medicine, possesses antibacterial, antiviral, and anti-inflammatory properties. The aim of this experiment was to investigate the therapeutic effect of Artemisia ordosica Krasch extraction (AOE) in treating colitis induced by dextran sulfate sodium (DSS) in mice. The in vitro antioxidant activity of AOE was evaluated by assessing its iron reduction capacity and scavenging capacity towards 1,1-diphenyl-2-picrylhydrazyl (DPPH) and hydroxyl radicals (·OH). The protective effect of AOE on colitis in mice was determined by monitoring key indicators such as body weight, colon length, and survival rate in mice, as well as by assessing the expression of colon-related genes and cytokine levels. We evaluated the impact of AOE on intestinal microbiota by measuring the 16s sequencing of cecal contents and bile acid metabolism. The results showed that the iron reduction capacity of AOE was positively correlated with its concentration. The half-maximal inhibitory concentrations (IC50) for scavenging DPPH and hydroxyl radicals were 3.126 mg/mL and 6.139 mg/mL, with a 95% confidence interval of 95%. In vivo studies demonstrated that AOE reduced DSS-induced colitis in mice by increasing the colon length, enhancing antioxidant enzyme activity, inhibiting inflammatory cell infiltration, suppressing the formation of TNF-α and IL-6, and reducing malondialdehyde (MDA) levels. qPCR analysis revealed that AOE reversed the down-regulation of Claudin mRNA expression, and altered the composition of cecal microbiota, thus mitigating DSS-induced colitis. AOE plays a crucial role in alleviating colitis in mice and effectively improves DSS-induced colitis, highlighting its potential as a therapeutic agent for inflammatory bowel diseases.

Antioxidants doi: 10.3390/antiox14010043

Authors: Paola Venditti Gaetana Napolitano

Mitochondria are found, with rare exceptions [...]

Antioxidants doi: 10.3390/antiox14010042

Authors: Blanca Martínez-Inda Nerea Jiménez-Moreno Irene Esparza Carmen Ancín-Azpilicueta

Cocoa and coffee are two of the world’s most important crops. Therefore, their by-products are generated in large quantities. This work proposes a simple method for the valorization of these residues by obtaining phenolic compounds and melanoidins by solid–liquid extraction using different hydroalcoholic solutions as extracting solvents (0, 25, 50, 75, 100% ethanol). Extracts of both by-products presented the highest antioxidant capacity and total phenolic and melanoidin content when using 50–75% ethanol in the solvent. Among all the extracts, those obtained from spent coffee grounds at 75% ethanol showed the highest concentrations of total phenolic compounds (13.5 ± 1.3 mmol gallic acid equivalents/g dry matter) and melanoidins (244.4 ± 20.1 mg/g dry matter). Moreover, the sun protection factor values of the coffee extracts obtained with 50 and 75% of ethanol as extraction solvent (7.8 ± 0.9 and 8.5 ± 0.7, respectively) showed their potential for use in the cosmetic sector. The most important phenolic compounds identified in the coffee by-products extracts were phenolic acids, and most of them were found in higher concentration in extracts obtained with lower percentages of ethanol (0–25%). Protocatechuic acid was the most abundant phenolic in cocoa extracts, with concentrations ranging from 18.49 ± 2.29 to 235.35 ± 5.55 µg/g dry matter, followed by 4-hydroxybenzoic acid, (-)-epicatechin and (+)-catechin. Esculetin was found in both coffee and cocoa extracts, which had not been reported to date in these residues. In summary, the use of 75% ethanol as an extraction solvent seems a good strategy to obtain extracts rich in phenolic compounds from food by-products rich in melanoidins, such as coffee and cocoa by-products. The high antioxidant potential of these extracts makes them of great interest for the cosmetic and nutraceutical industries.

Antioxidants doi: 10.3390/antiox14010041

Authors: Zhen Yang Jingyuan Man Haoyu Liu Di Wu Qiangwen Gu Hongjuan Zhang Yu Liu Dan Shao Baocheng Hao Shengyi Wang

Oxidative stress refers to the phenomenon in which the redox balance of the body is disrupted in response to stimuli, leading to an excessive accumulation of reactive oxygen species in vivo, which can lead to a variety of diseases. In contrast to artificial antioxidants, whose safety is controversial, natural antioxidants, which are widely available, pharmacologically active, and have little toxic side effects, are expected to be candidates for the treatment of oxidative stress-related diseases. Polygonum viviparum L. (PV) is a natural herbal medicine with antioxidant properties and is used as a traditional medicine in the Tibetan Plateau region. However, there are few studies that have focused on its antioxidant activity and mechanism of action in vitro and in vivo. Therefore, the present study firstly demonstrated that PV could exert good in vitro antioxidant effects by scavenging DPPH radicals and inhibiting the production of hydroxyl radicals through in vitro experiments. Secondly, PV was proven to attenuate the effects of oxidative stress on body weight gain and thymus development by establishing the Senna leaf-induced diarrhea model in rats, as well as to increase the activity of antioxidant enzymes and the content of non-enzymatic antioxidants in the intestinal tract and to enhance the rats’ own antioxidant defenses, to mitigate the oxidative damage caused by diarrhea. Subsequently, the application of the cellular oxidative stress model evidenced that PV could play a protective role against cellular oxidative stress by inhibiting the overaccumulation of ROS in macrophages. Furthermore, the candidate antioxidant targets of PV were analyzed and screened using a comprehensive network pharmacology method, and their expression were then examined at the mRNA level and protein level. Our results suggest that PV may protect against H2O2-induced oxidative damage in macrophages by activating BCL2L1 and inhibiting ESR1, JAK2/STAT3, and MMP2. These findings open new perspectives on the antioxidant mechanism of PV and the prospect of developing it as a novel natural antioxidant drug.

Antioxidants doi: 10.3390/antiox14010040

Authors: Nisa Beril Sen Irena Vovk Hasan Kırmızıbekmez Etil Guzelmeric

Qualitative and quantitative differences in the chemical composition between bee pollen originated from Castanea sativa (Türkiye and Slovenia), Salix spp. (Türkiye and Slovenia), and Quercus spp. (Türkiye) and androecia of Castanea sativa, Salix alba, and Quercus pubescens (apetalous trees) were evaluated for the first time by new high-performance thin-layer chromatography (HPTLC) and ultra-performance liquid chromatography (UPLC) methods using marker compounds. N1,N5,N10-tricaffeoylspermidine was isolated, and its structure was elucidated by nuclear magnetic resonance (NMR) and high-resolution mass spectrometry (HRMS). It was the main and the marker compound common to bee pollen (≈3–41 mg/g) and androecia (≈3–6 mg/g) samples. To the best of our knowledge, this is the first report of the identification of N1,N5,N10-tricaffeoylspermidine in bee pollen originated from Salix spp. and androecia of C. sativa, S. alba, and Q. pubescens. The botanical origins of bee pollen were determined via phytochemical profiling using HPTLC-image analyses showing that bee pollen from the same botanical source had almost identical profiles regardless of collection location, geographical differences, and the bee race. In vitro tests and HPTLC-effect-directed analyses (EDAs) were performed to assess antioxidant and xanthine oxidase (XO) inhibitory activities of bee pollen, androecia, and N1,N5,N10-tricaffeoylspermidine. HPTLC-EDA combined with image analyses was used for comparing the activities of bee pollen, androecia, N1,N5,N10-tricaffeoylspermidine, and also other marker compounds (quercetin, myricitrin, hyperoside, quercitrin, isoquercitrin, and rutin). The remarkable bioactivity of N1,N5,N10-tricaffeoylspermidine was for the first time evaluated by HPTLC-EDA and in vitro tests. This is the first study performing HPTLC-XO inhibitory activity analyses on the HPTLC NH2 F254S plates. Further bioactivity studies on botanically and chemically well-characterized bee pollen samples are needed to aid in the use of bee pollen-containing supplements in the prevention and treatment of diseases.

Antioxidants doi: 10.3390/antiox14010039

Authors: Mehboob Ahmed Umair Riaz Haimiao Lv Muhammad Amjad Sohail Ahmed Shaokat Ali Muhammad Usman Ghani Guohua Hua Liguo Yang

Inflammation disrupts the normal function of granulosa cells (GCs), which leads to ovarian dysfunction and fertility decline. Inflammatory conditions such as polycystic ovary syndrome (PCOS), primary ovarian insufficiency (POI), endometriosis, and age-related ovarian decline are often associated with chronic low-grade inflammation. Nicotinamide mononucleotide (NMN) is an important precursor of NAD+ and has gained attention for its potential to modulate cellular metabolism, redox homeostasis, and mitigate inflammation. This study investigated the protective roles of NMN against lipopolysaccharide LPS-mediated inflammation in GCs. The results of this experiment demonstrated that LPS had negative effects on GCs in term of reduced viability and proliferation rates and upregulated the production of pro-inflammatory cytokines, including interleukin-1 beta (IL-1β), interleukin-6 (IL-6), cyclooxygenase-2 (Cox-2), and tumor necrosis factor-alpha (TNF-α). Notably, the levels of NAD+ and NAD+/NADH ratio in GCs were reduced in response to inflammation. On the other hand, NMN supplementation restored the NAD+ levels and the NAD+/NADH ratio in GCs and significantly reduced the expression of pro-inflammatory markers at both mRNA and protein levels. It also enhanced cell viability and proliferation rates of GCs. Furthermore, NMN also reduced apoptosis rates in GCs by downregulating pro-apoptotic markers, including Caspase-3, Caspase-9, and Bax while upregulating anti-apoptotic marker Bcl-2. NMN supplementation significantly reduced reactive oxygen species ROS and improved steroidogenesis activity by restoring the estradiol (E2) and progesterone (P4) levels in LPS-treated GCs. Mechanistically, this study found that NMN suppressed the activation of the TLR4/NF-κB/MAPK signaling pathways in GCs, which regulates inflammatory processes. In conclusion, the findings of this study revealed that NMN has the potential to reduce LPS-mediated inflammatory changes in GCs by modulating NAD+ metabolism and inflammatory signaling pathways. NMN supplementation can be used as a potential therapeutic agent for ovarian inflammation and related fertility disorders.

Antioxidants doi: 10.3390/antiox14010038

Authors: Zhaoshan Zhang Jiawei Guo

Oxidative stress plays a pivotal role in the pathogenesis and progression of cardiovascular diseases (CVDs). This review focuses on the signaling pathways of oxidative stress during the development of CVDs, delving into the molecular regulatory networks underlying oxidative stress in various disease stages, particularly apoptosis, inflammation, fibrosis, and metabolic imbalance. By examining the dual roles of oxidative stress and the influences of sex differences on oxidative stress levels and cardiovascular disease susceptibility, this study offers a comprehensive understanding of the pathogenesis of cardiovascular diseases. The study integrates key findings from current research in three comprehensive ways. First, it outlines the major CVDs associated with oxidative stress and their respective signaling pathways, emphasizing oxidative stress’s central role in cardiovascular pathology. Second, it summarizes the cardiovascular protective effects, mechanisms of action, and animal models of various antioxidants, offering insights into future drug development. Third, it discusses the applications, advantages, limitations, and potential molecular targets of gene therapy in CVDs, providing a foundation for novel therapeutic strategies. These tables underscore the systematic and integrative nature of this study while offering a theoretical basis for precision treatment for CVDs. A major contribution of this study is the systematic review of the differential effects of oxidative stress across different stages of CVDs, in addition to the proposal of innovative, multi-level intervention strategies, which open new avenues for precision treatment of the cardiovascular system.

Antioxidants doi: 10.3390/antiox14010037

Authors: Hee-Jin Song Ayun Seol Jumin Park Ji-Eun Kim Tae-Ryeol Kim Ki-Ho Park Eun-Seo Park Su-Jeong Lim Su-Ha Wang Ji-Eun Sung Youngwoo Choi Heeseob Lee Dae-Youn Hwang

Oxidative stress is the key cause of the etiopathogenesis of several diseases associated with constipation. This study examined whether the green pine cone can improve the symptoms of constipation based on the antioxidant activities. The changes in the key parameters for the antioxidant activity and laxative effects were examined in the loperamide (Lop)-induced constipation of Sprague-Dawley (SD) rats after being treated with the methanol extracts of green pine cone (MPC, unripe fruits of Pinus densiflora). MPC contained several bioactive compounds, including diterpenoid compounds such as dehydroabietic acid, taxodone, and ferruginol. In addition, it exhibited high scavenging activity against 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2′-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radicals. These effects of MPC successfully reflected the improvement in nicotinamide adenine dinucleotide phosphate oxidase (NADP) H oxidase transcription, superoxide dismutase (SOD) levels, and nuclear factor erythroid 2-related factor 2 (Nrf2) phosphorylation levels in the mid colon of Lop+MPC-treated SD rats. Furthermore, significant improvements in the stool parameters, gastrointestinal (GI) transit, intestine length, and histopathological structure of the mid colon were detected in the Lop-induced constipation rats after MPC treatment. The other parameters, including the regulators for the adherens junction (AJ) and tight junction (TJ), and GI hormone secretion for laxative effects, were improved significantly in Lop+MPC-treated SD rats. These effects were also verified in Lop+MPC-treated primary rat intestine smooth muscle cells (pRISMCs) through analyses for antioxidant defense mechanisms. Overall, the finding of this study offers novel scientific evidence that MPC could be considered as a significant laxative for chronic constipation based on its antioxidant activity.

Antioxidants doi: 10.3390/antiox14010036

Authors: Dala N. Daraghmeh Rafik Karaman

Red blood cells (RBCs) are a vital component of the body’s oxygen supply system. In addition to being pro-oxidants, they are also essential components of the body’s antioxidant defense mechanism. RBCs are susceptible to both endogenous and exogenous sources of oxidants. Oxyhemoglobin autoxidation is the primary source of endogenous RBC oxidant production, which produces superoxide radicals and hydrogen peroxide. Potent exogenous oxidants from other blood cells and the surrounding endothelium can also enter RBCs. Both enzymatic (like glutathione peroxidase) and non-enzymatic (like glutathione) mechanisms can neutralize oxidants. These systems are generally referred to as oxidant scavengers or antioxidants, and they work to neutralize these harmful molecules (i.e., oxidants). While their antioxidative capabilities are essential to their physiological functions and delivering oxygen to tissues, their pro-oxidant behavior plays a part in several human pathologies. The redox-related changes in RBCs can have an impact on their function and fate. The balance between pro-oxidants and antioxidants determines the oxidative status of cells, which affects signal transduction, differentiation, and proliferation. When pro-oxidant activity exceeds antioxidative capacity, oxidative stress occurs, leading to cytotoxicity. This type of stress has been linked to various pathologies, including hemolytic anemia. This review compiles the most recent literature investigating the connections between RBC redox biochemistry, antioxidants, and diverse disorders.

Antioxidants doi: 10.3390/antiox14010035

Authors: Jie Ding Jiahui Liu Qingqi Guo Na Zhang

This study investigates the anti-aging effects of various concentrations of blueberry anthocyanins (BA) on the lifespan and health-related phenotypes of Caenorhabditis elegans. Blueberry anthocyanins were administered at concentrations of 50.0 μg/mL, 200.0 μg/mL, and 500.0 μg/mL, and their effects on nematode lifespan, locomotion, pharyngeal pumping rate, and the accumulation of lipofuscin and reactive oxygen species (ROS) were examined. Transcriptomic analysis was conducted to explore the regulatory effects of BA on anti-aging molecular pathways and key genes in C. elegans. Results showed a significant, dose-dependent extension of lifespan, improvement in locomotion and pharyngeal pumping rate, and reduction in lipofuscin and ROS accumulation. Transcriptomic analysis revealed that BA activated anti-aging pathways such as FOXO, IIS, and PI3K/Akt, upregulating critical genes like daf-16. These findings highlight the potential of blueberry anthocyanins as promising anti-aging agents through multiple physiological and molecular mechanisms.

Antioxidants doi: 10.3390/antiox14010034

Authors: Yu Cai Hua Yang Hui Xu Shanglai Li Bingru Zhao Zhibo Wang Xiaolei Yao Feng Wang Yanli Zhang

Oxidative stress is a significant factor in the death of granulosa cells (GCs), leading to follicular atresia and consequently limiting the number of dominant follicles that can mature and ovulate within each follicular wave. Follicular fluid contains a diverse array of metabolites that play crucial roles in regulating GCs’ proliferation and oocyte maturation, which are essential for follicle development and female fertility. However, the mechanisms behind metabolite heterogeneity and its effects on GCs’ function remain poorly understood. Here, we identified elevated nicotinamide levels in the follicular fluid of high-prolificacy sheep, correlated with oxidative stress in GCs, by an integrated analysis. In vitro experiments demonstrated that supplementation with β-nicotinamide mononucleotide (NMN) significantly increased the levels of nicotinamide adenine dinucleotide (NAD+) and adenosine triphosphate (ATP) in GCs. NMN treatment effectively reduced Lipopolysaccharide (LPS)-induced apoptosis and mitigated mitochondrial dysfunction, while also decreasing the production of reactive oxygen species (ROS), thereby enhancing the activity of the antioxidant defense system. Importantly, NMN treatment improved the impairments in steroid hormone levels induced by LPS. Mechanistically, the protective effects of NMN against GCs function were mediated via the AMPK/mTOR pathway. Collectively, our findings elucidate the metabolic characteristics associated with sheep prolificacy and demonstrate that NMN effectively protects GCs from LPS-induced dysfunction and enhances ovarian responsiveness via the AMPK/mTOR pathway. These findings also position NMN as a potential novel metabolic biomarker in enhancing ovarian function.

Antioxidants doi: 10.3390/antiox14010033

Authors: Yue Yan Changhe Wei Xin Liu Xin Zhao Shanmei Zhao Shuai Tong Guoyou Ren Qin Wei

The volatility, instability, and water insolubility of Camphora longepaniculata essential oil (CLEO) limit its practical applications in the food, pharmaceutical, and cosmetics industries. CLEO nanoemulsions (CLNEs) were formulated and characterized to overcome the aforementioned issues. The volatile compounds of CLEO were identified by gas chromatography–mass spectrometry. CLNEs were prepared using EL-40 (5%, w/w) as the surfactant via the high-pressure homogenization method. The formation of nanoemulsions was verified by Fourier transform infrared spectroscopy and transmission electron microscopy. Homogenized nanoemulsions had smaller particle sizes of 39.99 ± 0.47 nm and exhibited enhanced stability. The nanostructured CLEO showed an antibacterial effect against Escherichia coli and Staphylococcus aureus. The antibacterial mechanism was explored through bacterial morphology and intracellular lysate leakage. CLNEs disrupted the structure of bacterial cells and impaired the permeability of cell membranes, resulting in the leakage of bacterial intracellular contents. The nanoemulsions exhibited superior radical scavenging ability compared to the pure oil. Furthermore, evaluations of the cellular safety of the CLNEs demonstrated that the survival rate of exposed HOECs was greater than 90%, with an apoptosis rate of less than 10% in a concentration range. The results demonstrated that nanoemulsification improved the stability, solubility in aqueous media, and bioavailability of CLEO, thereby broadening its potential industrial applications as a natural antibacterial and antioxidant agent.

Antioxidants doi: 10.3390/antiox14010032

Authors: Angelika Szymańska Anna Nowak Anna Lipert Ewa Kochan

Ginseng has multi-directional pharmacological properties. Some data suggest that ginseng can enhance physical endurance, which, in turn, leads to protection of the cardiovascular system. However, not all experiments are conclusive. For this reason, the main aim of this research was to perform a meta-analysis and review of studies published between the years 2013 and 2023 concerning the ginseng effect on physical performance in animal and human models. Medline, Pubmed, and ClinicalKey electronic databases were used to analyze data. The search strategy included the following criteria: ginseng and exercise; ginseng supplementation; and ginseng supplements. The results suggest that ginseng supplementation may have a positive effect on CK levels in animal studies. Similar observations were stated in relation to serum lactate and BUN. Furthermore, a human study showed a significant increase in exercise time to exhaustion and VO2 max after supplementation. The review of the literature and conducted meta-analysis identified that ginseng supplementation may have a positive effect on exercise endurance. Due to the fact that most of the current studies were based on animal models, further research on human models is needed to identify the most effective dosage or form of applied ginseng to be a supportive element in CVD management.

Antioxidants doi: 10.3390/antiox14010031

Authors: Maja Molnar Martina Jakovljević Kovač Lidija Jakobek Lovro Mihajlović Valentina Pavić

This study explores the green extraction of phenolic antioxidants from Aronia melanocarpa fruit using choline-chloride-based deep eutectic solvents (DESs) as an eco-friendly alternative to conventional solvents. Sixteen DESs, prepared by combining choline chloride with various hydrogen bond donors, were characterized for their physical properties, including viscosity, polarity, and pH, and applied to extract phenolics from Aronia melanocarpa. High-performance liquid chromatography (HPLC) quantified key phenolic compounds, including neochlorogenic and chlorogenic acid, quercetin derivatives, and cyanidin derivatives, as well as total phenolic acids, flavanols, and anthocyanins. The results revealed that DES composition and physical properties significantly influenced extraction efficiency and antioxidant activity. Additionally, the intrinsic antioxidant activity of DESs contributed substantially to the overall activity of the extracts, particularly in DESs containing organic acids or thiourea. Choline chloride/tartaric acid DES demonstrated the highest total phenolic content, attributed to its high viscosity and strongly acidic pH, while choline chloride/thiourea DES, with low viscosity and slightly acidic pH, exhibited the greatest antioxidant activity. This study highlights how tailoring DES formulations can optimize the extraction of target compounds while accounting for the solvent’s intrinsic properties. The findings support the potential application of DESs as environmentally friendly solvents in the food, pharmaceutical, and cosmetic industries.

Antioxidants doi: 10.3390/antiox14010030

Authors: Kai Wang Ling Qin Huan Lin Mengke Yao Junhan Cao Qing Zhang Changfeng Qu Yingying He Jinlai Miao Ming Liu

Mycosporine-glycine (M-Gly), a member of the mycosporine-like amino acid (MAA) family, is known for its potent antioxidant and anti-inflammatory properties. However, its in vivo efficacy in alleviating acute skin photodamage, primarily caused by oxidative stress, has not been well explored. In this investigation, 30 female ICR mice were divided into four groups: a control group and three Ultraviolet B (UVB)-exposed groups treated with saline or M-Gly via intraperitoneal injection for 30 days. At the end of the experiment, UVB exposure caused erythema, wrinkling, collagen degradation, and mast cell infiltration in mouse dorsal skin. M-Gly treatment improved skin appearance and reduced mast cell numbers, while also elevating antioxidant levels, including superoxide dismutase (SOD), catalase (CAT), and glutathione (GSH). Furthermore, M-Gly reduced inflammatory cytokines, such as tumor necrosis factor-alpha (TNF-α), interleukin-6 (IL-6), and IL-1β, typically upregulated after UVB exposure. M-Gly also protected skin collagen by upregulating type I procollagen and decreasing MMP-1 levels. Skin metabolomic profiling identified 34 differentially abundant metabolites, and transcriptomic analysis revealed 752 differentially expressed genes. The combined metabolomic and transcriptomic data indicate that M-Gly’s protective effects may involve the regulation of ion transport, cellular repair, metabolic stability, collagen preservation, and the Nrf2/HO-1 pathway. These findings highlight M-Gly’s potential as an endogenous antioxidant for protecting skin from UVB-induced damage.

Antioxidants doi: 10.3390/antiox14010029

Authors: George J. Kontoghiorghes

Epidemiological studies have suggested that following long-term, low-dose daily aspirin (LTLDA) administration for more than 5 years at 75–100 mg/day, 20–30% of patients (50–80 years old) had a lower risk of developing colorectal cancer (CRC) and about the same proportion in developing iron deficiency anemia (IDA). In cases of IDA, an increase in iron excretion is suspected, which is caused by aspirin chelating metabolites (ACMs): salicylic acid, salicyluric acid, 2,5-dihydroxybenzoic acid, and 2,3-dihydroxybenzoic acid. The ACMs constitute 70% of the administered aspirin dose and have much longer half-lives than aspirin in blood and tissues. The mechanisms of cancer risk reduction in LTLDA users is likely due to the ACM’s targeting of iron involved in free radical damage, iron-containing toxins, iron proteins, and associated metabolic pathways such as ferroptosis. The ACMs from non-absorbed aspirin (about 30%) may also mitigate the toxicity of heme and nitroso-heme and other iron toxins from food, which are responsible for the cause of colorectal cancer. The mode of action of aspirin as a chelating antioxidant pro-drug of the ACMs, with continuous presence in LTLDA users, increases the prospect for prophylaxis in cancer and other diseases. It is suggested that the anticancer effects of aspirin depend primarily on the iron-chelating antioxidant activity of the ACMs. The role of aspirin in cancer and other diseases is incomplete without considering its rapid biotransformation and the longer half-life of the ACMs.

Antioxidants doi: 10.3390/antiox14010028

Authors: Hangfan Li Zibin Wu Baisheng Yu Jinyuan Chen Chuang Yang Yongqing Guo Baoli Sun

This study investigated the effects of dietary capsaicin supplementation on antioxidant capacity, immune function, and gut microbiota in periparturient dairy cows. Twenty Holstein cows with an average parity of 2.5 ± 0.76, milk production of 31.30 ± 2.39 kg, and 36.10 ± 2.38 days to calving were randomly assigned to either a control group fed a basal diet or a treatment group supplemented with 1.2 g/head/day of capsaicin. The supplementation was administered during an evaluation period spanning from 28 days before delivery to 21 days after delivery using a randomized block experimental design. Results showed that capsaicin significantly reduced milk somatic cell count and pro-inflammatory cytokines (TNF-α, IL-1β, and IL-6) while enhancing serum antioxidant enzymes (SOD, GSH-Px, and CAT) and immunoglobulin levels (IgG, IgA, and IgM). Moreover, capsaicin altered gut microbiota composition, increasing the relative abundance of beneficial genera. These findings suggest that dietary capsaicin supplementation during the transition period improves lactation performance and supports immune function, as well as alleviates oxidative stress. This study highlights the potential of capsaicin as a practical dietary strategy for enhancing productivity in dairy farming.

Antioxidants doi: 10.3390/antiox14010027

Authors: Yi Xue Tao Wang Ji-Peng Liu Qi Chen Xiao-Long Dai Min Su Yu-Hang Cheng Cheng-Chao Chu Yun-Qing Ren

Skin is a vital barrier for the human body, protecting against external environmental influences and maintaining internal homeostasis. In addition, an imbalance of oxidative stress and antioxidant mechanisms can lead to skin-related diseases. Thus, for treating skin-related diseases, antioxidant therapy may be an important strategy to alleviate these symptoms. However, traditional drug therapies have limitations in treating these conditions, such as lack of lasting effect and insufficient skin permeability. Recently, nano-antioxidants, with their good permeability, sustained-release ability, multifunctionality, and other beneficial characteristics, have showed their advances in the exploration of skin-related diseases from research on safe therapies to clinical practice. Hereby, we review the latest research and advancements in nano-antioxidants for skin-related diseases. We categorize skin-related diseases into four main groups: skin inflammatory diseases, skin damage caused by ultraviolet rays, skin wound healing, and other skin-related conditions. Additionally, we summarize the prospects and potential future directions for nano-antioxidant drugs in treating skin-related diseases.

Antioxidants doi: 10.3390/antiox14010026

Authors: Ines Angelini Mariangela Centrone Giusy Rita Caponio Annarita Di Mise Andrea Gerbino Marianna Ranieri Giovanna Valenti Grazia Tamma

The water channel AQP3 is an aquaglyceroporin expressed in villus epithelial cells, and it plays a role in water transport across human colonic surface cells. Beyond water, AQP3 can mediate glycerol and H2O2 transport. Abnormal expression and function of AQP3 have been found in various diseases often characterized by altered cell growth and proliferation. Here, the beneficial effects of MOMAST® have been evaluated. MOMAST® is an antioxidant-patented natural phenolic complex obtained from olive wastewater (OWW) of the Coratina cultivar. Treatment of human colon HCT8 cells with MOMAST® reduced cell viability. Confocal studies and Western Blotting analysis demonstrated that treatment with MOMAST® significantly decreased the staining and the expression of AQP3. Importantly, functional studies revealed that the reduction of AQP3 abundance correlates with a significant decrease in glycerol and H2O2 uptake. Indeed, the H2O2 transport was partially but significantly reduced in the presence of MOMAST® or DFP00173, a selective inhibitor of AQP3. In addition, the MOMAST®-induced AQP3 decrease was associated with reduced epithelial-mesenchymal transition (EMT)-related proteins such as vimentin and β-catenin. Together, these findings propose MOMAST® as a potential adjuvant in colon diseases associated with abnormal cell growth by targeting AQP3.

Antioxidants doi: 10.3390/antiox14010025

Authors: Mariusz Sieminski Michalina Reimus Maria Kałas Ewelina Stępniewska

Traumatic brain injury (TBI) is a disease resulting from external physical forces acting against the head, leading to transient or chronic damage to brain tissue. Primary brain injury is an immediate and, therefore, rather irreversible effect of trauma, while secondary brain injury results from a complex cascade of pathological processes, among which oxidative stress and neuroinflammation are the most prominent. As TBI is a significant cause of mortality and chronic disability, with high social costs all over the world, any form of therapy that may mitigate trauma-evoked brain damage is desirable. Melatonin, a sleep–wake-cycle-regulating neurohormone, exerts strong antioxidant and anti-inflammatory effects and is well tolerated when used as a drug. Due to these properties, it is very reasonable to consider melatonin as a potential therapeutic molecule for TBI treatment. This review summarizes data from in vitro studies, animal models, and clinical trials that focus on the usage of melatonin in TBI.

Antioxidants doi: 10.3390/antiox14010024

Authors: Yaoxiang Li Shivani Bansal Baldev Singh Meth M. Jayatilake William Klotzbier Marjan Boerma Mi-Heon Lee Jacob Hack Keisuke S. Iwamoto Dörthe Schaue Amrita K. Cheema

Exposure to ionizing radiation disrupts metabolic pathways and causes oxidative stress, which can lead to organ damage. In this study, urinary metabolites from mice exposed to high-dose and low-dose whole-body irradiation (WBI HDR, WBI LDR) or partial-body irradiation (PBI BM2.5) were analyzed using targeted and untargeted metabolomics approaches. Significant metabolic changes particularly in oxidative stress pathways were observed on Day 2 post-radiation. By Day 30, the WBI HDR group showed persistent metabolic dysregulation, while the WBI LDR and PBI BM2.5 groups were similar to control mice. Machine learning models identified metabolites that were predictive of the type of radiation exposure with high accuracy, highlighting their potential use as biomarkers for radiation damage and oxidative stress.

Antioxidants doi: 10.3390/antiox14010023

Authors: Aekkhaluck Intharuksa Warunya Arunotayanun Mingkwan Na Takuathung Yaowatat Boongla Siripat Chaichit Suthiwat Khamnuan Anchalee Prasansuklab

Particulate matter (PM), particularly fine (PM2.5) and ultrafine (PM0.1) particles, originates from both natural and anthropogenic sources, such as biomass burning and vehicle emissions. These particles contain harmful compounds that pose significant health risks. Upon inhalation, ingestion, or dermal contact, PM can penetrate biological systems, inducing oxidative stress, inflammation, and DNA damage, which contribute to a range of health complications. This review comprehensively examines the protective potential of natural products against PM-induced health issues across various physiological systems, including the respiratory, cardiovascular, skin, neurological, gastrointestinal, and ocular systems. It provides valuable insights into the health risks associated with PM exposure and highlights the therapeutic promise of herbal medicines by focusing on the natural products that have demonstrated protective properties in both in vitro and in vivo PM2.5-induced models. Numerous herbal medicines and phytochemicals have shown efficacy in mitigating PM-induced cellular damage through their ability to counteract oxidative stress, suppress pro-inflammatory responses, and enhance cellular defense mechanisms. These combined actions collectively protect tissues from PM-related damage and dysfunction. This review establishes a foundation for future research and the development of effective interventions to combat PM-related health issues. However, further studies, including in vivo and clinical trials, are essential to evaluate the safety, optimal dosages, and long-term effectiveness of herbal treatments for patients under chronic PM exposure.

Antioxidants doi: 10.3390/antiox14010022

Authors: Jianqin Zhang Dianfeng Zheng Naijie Feng Aaqil Khan Rui Deng Jian Xiong Linchong Ding Zhiyuan Sun Jiahuan Li Xiaohui Yang Chen Wu

Salt stress inhibits rice seed germination. Strigolactone (GR24) plays a vital role in enhancing plant tolerance against salt stress. However, GR24’s impact on the metabolism of stored substances and endogenous hormones remains unclear. This study investigated the impact of exogenous GR24 on the metabolism of stored substances and endogenous hormones during the early stages of rice seed germination under salt stress. The results showed that salt stress significantly reduced the germination rate, germination potential, germination index, radicle length, germ length, and fresh and dry weights of the radicle and germ under salt stress. Pre-treatment (1.2 μmol L−1 GR24) significantly reduced the inhibition of salt stress on rice seed germination and seedling growth. GR24 promoted the decomposition of starch by enhancing the activities of α-amylase, β-amylase, and total amylase and improved the levels of soluble sugars and proteins and the conversion rate of substances under salt stress. GR24 effectively enhanced the activities of superoxide dismutase (SOD), catalase (CAT), and ascorbate peroxidase (APX); increased ascorbic acid (ASA) and glutathione (GSH) levels; and reduced malondialdehyde (MDA) content. This reduced the oxidative damage of salt stress. Furthermore, GR24 significantly increased the contents of strigolactones (SLs), auxin (IAA), gibberellin (GA3), cytokinin (CTK) as well as IAA/ABA, CTK/ABA, GA/ABA, and SL/ABA ratios and reduced abscisic acid (ABA) levels. The current findings indicate that GR24 effectively mitigates the adverse impact salt stress by regulating antioxidant enzyme activity and endogenous hormone balance.

Antioxidants doi: 10.3390/antiox14010021

Authors: Radosław Dutczak Marita Pietrucha-Dutczak

Electroretinography (ERG) is a non-invasive technique for evaluating the retinal function in various ocular diseases. Its results are useful for diagnosing ocular disorders and assessing disease progression or treatment effectiveness. Since numerous studies are based on animal models, validating the ERG results from animals is pivotal. The first part of this paper presents basic information on the types of ERG tests used on rodents, and the second part describes the recorded functional changes in rodents’ retinas when various antioxidant treatments for diabetic retinopathy were used. Our study showed that among the tests for diabetic retinopathy diagnosis in rodents, full-field ERG is accurate and the most commonly used, and pattern ERG and the photopic negative response of the flash ERG tests are rarely chosen. Furthermore, antioxidants generally protect retinas from functional losses. Their beneficial influence is expressed in the preserved amplitudes of the a- and b-waves and the oscillatory potentials. However, prolonging the drug exposure showed that the antioxidants could delay the onset of adverse changes but did not stop them. Future studies should concentrate on how long-term antioxidant supplementation affects the retinal function.

Antioxidants doi: 10.3390/antiox14010020

Authors: Lucia Bartella Fabio Mazzotti Ilaria Santoro Ines Rosita Talarico Leonardo Di Donna

A novel and efficient analytical protocol based on paper spray tandem mass spectrometry was developed for the determination of isomeric O-glycoside flavanones in citrus juices and beverages. This approach significantly reduces sample preparation time and solvent consumption compared to traditional chromatographic techniques. By exploiting the unique fragmentation patterns of these compounds, accurate quantification of both diglycosides and their individual isomers (neohesperidoside and rutinose derivatives) was achieved. The method demonstrated excellent analytical performance, with high accuracy, selectivity, and reproducibility. The impact of matrix effects was mitigated through the construction of ratio calibration curves, ensuring reliable quantification in complex matrices. Finally, a simple DPPH experiment to assay the antioxidant activity of each single positional isomer was performed, indicating the superior ability of neohesperidose conjugates. This simplified method offers a valuable tool for quality control, authenticity assessment and the study of health benefits associated with citrus consumption.

Antioxidants doi: 10.3390/antiox14010019

Authors: Philippe Guerre Elodie Lassallette Amélie Guerre Didier Tardieu

This study aimed to assess the effects of a diet containing 20.8 mg FB1 + FB2/kg over four and nine days on oxylipin (OL) profiles in the liver and brain of chickens. A total of 96 OLs, derived from seven polyunsaturated fatty acids (PUFAs) via the cyclooxygenase (COX), lipoxygenase (LOX), cytochrome P450 (P450), and non-enzymatic pathways, were measured using HPLC-MS/MS. In the liver, a significant increase in epoxide P450-derived OLs was detected by day 4, with smaller but notable increases in COX- and LOX-derived OLs by day 9. These alterations were independent of whether the parent PUFA was ω6 or ω3. However, OLs derived from 18-carbon (C18) PUFAs, such as linoleic acid and alpha-linolenic acid, showed greater increases compared to those derived from C20 or C22 PUFAs. The diol/epoxide ratios in the liver decreased at four and nine days, suggesting that fumonisins did not induce an inflammatory response. In the brain, at four days, the most discriminative OLs were derived from ω3-PUFAs, including docosahexaenoic acid, docosapentaenoic acid, and alpha-linolenic acid, via the LOX pathway. By nine days, several OLs derived from arachidonic acid, spanning all enzymatic pathways, became discriminative. In general, the diol/epoxide ratios in the brain were decreased at 4 days and then returned to the initial levels. Taken together, these results show strong effects of fumonisins on OLs in the liver and brain that are both specific and distinct.

Antioxidants doi: 10.3390/antiox14010018

Authors: Laura Giannotti Benedetta Di Chiara Stanca Francesco Spedicato Daniele Vergara Eleonora Stanca Fabrizio Damiano Luisa Siculella

Cannabinoids include both endogenous endocannabinoids and exogenous phytocannabinoids, such as cannabidiol (CBD), and have potential as therapeutic agents in cancer treatment due to their selective anticancer activities. CBD exhibits both antioxidant and pro-oxidant effects depending on its concentration and cell types. These properties allow CBD to influence oxidative stress responses and potentially enhance the efficacy of antitumor therapies. In this study, we treated U87MG glioma cells with low dose (1 μM) CBD and evaluated its molecular effects. Our findings indicate that CBD reduced cell viability by 20% (p < 0.05) through the alteration of mitochondrial membrane potential. The alteration of redox status by CBD caused an attempt to rescue mitochondrial functionality through nuclear localization of the GABP transcription factor involved in mitochondria biogenesis. Moreover, CBD treatment caused an increase in autophagic flux, as supported by the increase in Beclin-1 and the ratio of LC3-II/LC3-I. Due to mitochondria functionality alteration, pro-apoptotic proteins were induced without activating apoptotic effectors Caspase-3 or Caspase-7. The study of the transcription factor NRF2 and the ubiquitin-binding protein p62 expression revealed an increase in their levels in CBD-treated cells. In conclusion, low-dose CBD makes U87MG cells more vulnerable to cytotoxic effects, reducing cell viability and mitochondrial dynamics while increasing autophagic flux and redox systems. This explains the mechanisms by which glioma cells respond to CBD treatment. These findings highlight the therapeutic potential of CBD, suggesting that modulating NRF2 and autophagy pathways could represent a promising strategy for glioblastoma treatment.

Antioxidants doi: 10.3390/antiox14010016

Authors: Diego Salagre Miguel Navarro-Alarcón Luis Gerardo González Mohamed A. Elrayess Marina Villalón-Mir Rocío Haro-López Ahmad Agil

Endoplasmic reticulum (ER) stress is a crucial factor in the progression of obesity-related type 2 diabetes (diabesity), contributing to skeletal muscle (SKM) dysfunction, calcium imbalance, metabolic inflexibility, and muscle atrophy. The ER and mitochondria together regulate intracellular calcium levels, and melatonin, a natural compound with antioxidant properties, may alleviate these challenges. Our previous research showed that melatonin raises intracellular calcium and preserves muscle structure by enhancing mitochondrial function in obese diabetic rats. This study further explores melatonin’s potential to reduce ER stress in the vastus lateralis (VL) muscle by modulating the unfolded protein response (UPR) and restoring calcium levels disrupted by diabesity. Five-week-old Zücker diabetic fatty (ZDF) rats and lean littermates of both sexes were divided into control and melatonin-treated groups (10 mg/kg/day for 12 weeks). Flame atomic absorption spectrometry results showed that melatonin restored VL intraorganellar calcium homeostasis, increasing calcium levels in mitochondria and reducing them in the ER by raising the activity and expression of calcium transporters in both sexes of ZDF rats. Melatonin also decreased ER stress markers (GRP78, ATF6, IRE1α, and PERK) and reduced pro-apoptosis markers (Bax, Bak, P-JNK, cleaved caspase 3 and 9) while increasing Bcl2 levels and melatonin receptor 2 (MT2) expression. These findings suggest that melatonin may protect against muscle atrophy in obese and diabetic conditions by mitigating ER stress and calcium imbalance, highlighting its therapeutic potential.

Antioxidants doi: 10.3390/antiox14010017

Authors: Yang-Jin Shen Yin-Cheng Huang Yi-Chuan Cheng

Spinal cord injury (SCI) initiates a cascade of secondary damage driven by oxidative stress, characterized by the excessive production of reactive oxygen species and other reactive molecules, which exacerbate cellular and tissue damage through the activation of deleterious signaling pathways. This review provides a comprehensive and critical evaluation of recent advancements in antioxidant-based therapeutic strategies for SCI, including natural compounds, RNA-based therapies, stem cell interventions, and biomaterial applications. It emphasizes the limitations of single-regimen approaches, particularly their limited efficacy and suboptimal delivery to injured spinal cord tissue, while highlighting the synergistic potential of combination therapies that integrate multiple modalities to address the multifaceted pathophysiology of SCI. By analyzing emerging trends and current limitations, this review identifies key challenges and proposes future directions, including the refinement of antioxidant delivery systems, the development of multi-targeted approaches, and strategies to overcome the structural complexities of the spinal cord. This work underscores the pressing need for innovative and integrative therapeutic approaches to advance the clinical translation of antioxidant-based interventions and improve outcomes for SCI patients.

Antioxidants doi: 10.3390/antiox14010015

Authors: Jinsick Kim Min Young Go Chae Young Jeon Jung Un Shin Mujun Kim Hye Won Lim Dong Wook Shin

Diabetic foot ulcers represent a severe complication of diabetes, often resulting in amputation and high mortality rates. Currently, there are no treatments for diabetic foot ulcers other than antibiotics and dressings. In this study, we evaluated the wound-healing effects of an antidiabetic agent pinitol in lipopolysaccharide (LPS)-damaged human dermal fibroblasts (HDFs) and streptozotocin (STZ)-induced diabetic rat models with a foot wound. Our findings indicated that pinitol enhanced cell migration, proliferation, and wound healing by activating Nrf2, thereby mitigating oxidative stress and inflammatory responses at the wound site. Additionally, pinitol restored mitochondrial energy metabolism, decreased matrix metalloproteinase (MMP) activity, and increased collagen deposition. Furthermore, pinitol facilitated angiogenesis, contributing to improved wound healing. Taken together, these findings suggest that pinitol could be a promising therapeutic agent for the treatment of diabetic foot ulcers.

Antioxidants doi: 10.3390/antiox14010014

Authors: Jie Li Haonan Zheng Jiahui Liu Jie Ding Qingqi Guo Na Zhang

Constipation is a prevalent global health issue that greatly affects human well-being. However, many existing treatments are associated with side effects, necessitating the development of alternative approaches. In this study, a balanced fatty acid red pine seed direct-drinking oil (SFA:MUFA:PUFA = 1.14:1.08:1, n − 6:n − 3 = 4.17:1) was formulated using red pine seed oil as the base oil, blended with coconut oil, rice bran oil, and camellia oil. The study investigated the effects and mechanisms of this red pine seed direct-drinking oil in alleviating constipation in mice. Results showed that, compared to normal mice, constipated mice exhibited symptoms of dry stools, difficulty defecating, abnormal neurotransmitter levels, oxidative stress, and colonic tissue damage. Additionally, the protein expression levels of occludin and claudin-1 were reduced by 86.11% and 25.00%, respectively (p < 0.05), while mRNA expression levels decreased by 70.80% and 59.00% (p < 0.05). Red pine seed direct-drinking oil intake improved defecation, reduced serum levels of vasoactive intestinal peptide (VIP), endothelin-1 (ET-1), and nitric oxide (NO), and increased substance P (SP) levels. Furthermore, it also significantly elevated serum levels of superoxide dismutase (SOD) and catalase (CAT) (p < 0.05), alleviated colonic tissue damage, and upregulated the protein and mRNA expression levels of occludin and claudin-1 (p < 0.05). These findings suggest that red pine seed direct-drinking oil alleviates constipation in mice by enhancing intestinal motility, regulating serum neurotransmitters, mitigating oxidative stress, repairing intestinal barrier damage, and increasing tight junction protein expression. This study represents the first use of red pine seed direct-drinking oil to alleviate constipation in mice, providing a novel approach to improving symptoms in individuals with constipation.

Antioxidants doi: 10.3390/antiox14010011

Authors: Yurong Li Juanjuan Wei Zimao Ye Chen Ji Wenji Li Li Xu Zhiqin Zhou

Citrus peel essential oils (CPEOs) have demonstrated substantial medicinal potential for glioblastoma treatment because of their extensive antitumor effects, low potential for drug resistance, and ability to cross the human blood–brain barrier. In this study, the chemical compositions of five CPEOs were analyzed via gas chromatography–mass spectrometry (GC-MS). CCK8 assays were used to evaluate the ability of five CPEOs to inhibit U251 human glioblastoma cells, and XLB and RA were selected for further investigation. Through wound healing assays and cell cycle and apoptosis analyses via flow cytometry, it was revealed that these CPEOs inhibited cell migration, arrested the cell cycle at G1/G0, and induced apoptosis with similar levels of inhibition. After CPEOs treatment, the intracellular Ca2+ content and reactive oxygen species levels in U251 cells increased significantly, whereas the mitochondrial membrane potential decreased. Additionally, the antioxidant enzyme system (SOD, POD, CAT, and GR) and the nonenzymatic defense system (GSH) were inhibited, leading to an increase in lipid peroxidation. qRT–PCR indicated the significant upregulation of intracellular calcium ion signaling pathways and the upregulation of mitochondrial apoptosis-related genes. Additionally, the activation of calcicoptosis-related indicators induced by the CPEOs could be reversed by inhibitor treatment, confirming that both of the selected CPEOs inhibit tumors by activating calcicoptosis-related pathways. These findings highlight the immense potential of CPEOs in healthcare and pharmaceutical applications by not only providing a scientific basis for the potential application of CPEOs in the treatment of glioblastoma but also offering new insights for the development of novel antitumor drugs.

Antioxidants doi: 10.3390/antiox14010013

Authors: Ah Young Yang Jung-Yeon Kim Mi-Gyeong Gwon Hyun Hee Kwon Jaechan Leem Eon-Ju Jeon

Chronic kidney disease (CKD) progresses through mechanisms involving inflammation, fibrosis, and oxidative stress, leading to the gradual structural and functional deterioration of the kidneys. Tormentic acid (TA), a triterpenoid compound with known anti-inflammatory and antioxidant properties, shows significant potential in counteracting these pathological processes. This study explored the protective role of TA in a unilateral ureteral obstruction (UUO)-induced CKD model. Mice received TA through intraperitoneal injections at a dosage of 5 mg/kg per day for 8 consecutive days, commencing a day before the UUO procedure. The TA treatment significantly improved both structural and functional kidney injury. It suppressed cytokine expression and reduced immune cell infiltration, inhibited the activation of the mitogen-activated protein kinase cascade, and alleviated endoplasmic reticulum stress. Moreover, TA displayed potent anti-fibrotic effects by reversing epithelial-to-mesenchymal transition and inhibiting Smad2/3 activation, reducing extracellular matrix deposition. TA also mitigated oxidative stress by attenuating lipid peroxidation and boosting antioxidant defenses. Additionally, it inhibited apoptosis and ferroptosis by reducing oxidative stress and modulating key cell death markers. Collectively, these findings indicate that TA provides comprehensive renoprotection in the UUO model by effectively targeting inflammation, fibrosis, oxidative stress, and tubular cell death in CKD progression.

Antioxidants doi: 10.3390/antiox14010012

Authors: Chuang Liu Jun Dang Minchen Wu

The exploration of drug targets has always been a priority in new drug research, and this work is even more essential for natural active compounds. Saxifraga tangutica is a traditional Tibetan medicine with excellent antioxidant properties. In this study, an alkaloid, N-p-coumaroyl-N’-caffeoylputrescine (PCC), was first isolated from the plant, Saxifraga tangutica, with a DPPH scavenging rate of 0.936 μg/mL. To further identify its target, the drug affinity responsive target stability technique and multiple public databases were integrated to retrieve a total of 317 common targets from comprehensive screening. A further bioinformatics analysis not only identified 13 hub targets but also indicated PCC as having biological activities against cancer and affecting metabolic diseases. Integrating reverse virtual docking, molecular dynamics simulations, and cellular thermal shift assays ultimately focused on HSP90AA1 as the target of PCC. An in vitro study on liver (HepG2) cells and breast (MCF-7) cancer cells revealed that PCC modulates HSP90AA1, subsequently affecting Mut-p53 expression, triggering a cascade effect that reduced adriamycin-induced drug resistance in cells. Furthermore, a prediction of the absorption, distribution, metabolism, excretion, and toxicity was also applied to evaluate the drug-like properties of PCC. Overall, the integrated strategy used in this study successfully identified the target of PCC, providing a valuable paradigm for future research on the action targets of natural products.

Antioxidants doi: 10.3390/antiox14010010

Authors: Jian Lyu Ning Jin Xianglan Ma Xueyun Yin Li Jin Shuya Wang Xuemei Xiao Jihua Yu

Free amino acids, polyphenols, and anthocyanins were quantified in 30 Chinese eggplant varieties. Moreover, antioxidant capacity characterizations including 2, 2’-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), ferric-reducing antioxidant power (FRAP), and 2,2-diphenyl-1-picrylhydrazyl (DPPH) were performed. The total amino acid content of the 30 eggplant varieties ranged from 15,267.19 to 26,827.4 mg kg−1 DW. The most abundant amino acids were glutamic acid, arginine, and aspartic acid. The coefficients of variation (CV) for the 20 amino acids ranged from 5.85 to 106.14%, of which 18 free amino acids had CVs > 20%. Total polyphenol and anthocyanin contents ranged from 17,097.41 to 39,474.98 µg g−1 DW and 5.28 to 978.32 µg g−1 DW, respectively. The variability of both polyphenol and anthocyanin components was >20%, with a range of 21.25–102.89%. Chlorogenic acid was the most abundant polyphenol. The total anthocyanin content of purple eggplant varieties was significantly higher than green varieties. Of the purple eggplant varieties, V28 (‘E150725’), V30 (‘1952’), and V16 (‘Weichangqie101’) had significantly higher total anthocyanins than the other eggplant varieties. DPPH, ABTS, and FRAP assays showed peaks at V3 (‘Zhengqie924’). Pearson’s correlation analysis revealed that polyphenols and anthocyanins were the main contributors to the antioxidant capacity of eggplants. A classification model with principal component analysis classified 30 Chinese eggplant varieties into two categories: high and low antioxidant capacities. The top five Chinese eggplant varieties ranked for amino acids, antioxidants, and antioxidant capacity were V29 (‘Zhengqie903’), V24 (‘Zhengqie78’), V1 (‘1871’), V3 (‘Zhengqie924’), and V28 (‘E150725’). These findings provide theoretical basis for high-quality breeding and producer/consumer selection of eggplants.

Antioxidants doi: 10.3390/antiox14010009

Authors: Ravinder Kumar Sherry Flint-Garcia Miriam Nancy Salazar Vidal Lakshmikantha Channaiah Bongkosh Vardhanabhuti Stephan Sommer Caixia Wan Pavel Somavat

Purple corn pericarp, a processing waste stream, is an extremely rich source of phytochemicals. Optimal polyphenol extraction parameters were identified using response surface methodology (RSM) by combining a deep eutectic solvent (DES) and ultrasound-assisted extraction (UAE) method. After DES characterization, Plackett–Burman design was used to screen five explanatory variables, namely, time, Temp (temperature), water, Amp (amplitude), and S/L (solid-to-liquid ratio). The total anthocyanin concentration (TAC), total polyphenol concentration (TPC), and condensed tannin (CT) concentration were the response variables. After identifying significant factors, the Box–Behnken design was utilized to identify the optimal extraction parameters. The experimental yields under the optimized conditions of time (10 min), temperature (60 °C), water concentration (42.73%), and amplitude (40%) were 36.31 ± 1.54 g of cyanidin-3-glucoside (C3G), 103.16 ± 6.17 g of gallic acid (GA), and 237.54 ± 9.98 g of epicatechin (EE) per kg of pericarp, with a desirability index of 0.858. The relative standard error among the predicted and experimental yields was <10%, validating the robustness of the model. HPLC analysis identified seven phytochemicals, and significant antioxidant activities were observed through four distinct assays. Metabolomic profiling identified 57 unique phytochemicals. The UAE technique combined with DES can efficiently extract polyphenols from purple corn pericarp in a short time.

Antioxidants doi: 10.3390/antiox14010008

Authors: Chia-Hsuan Tsai Ya-Lun Liou Sin-Min Li Hsiang-Ruei Liao Jih-Jung Chen

Rhamnus formosana is a creeping evergreen shrub endemic to Taiwan. In traditional medicine, Rhamnaceae plants are used as herbal remedies for conditions such as itching, difficulty urinating, and constipation. This study explores the inhibitory effects of various solvent extracts and bioactive components of R. formosana on α-glucosidase, tyrosinase, acetylcholinesterase (AChE), and antioxidant activity. The 100 °C water extract exhibited strong antioxidant activity in DPPH, ABTS, superoxide, and FRAP assays. The methanol extract demonstrated the highest α-glucosidase inhibitory effect, while the ethanol extract displayed potent AChE inhibition and the acetone extract showed the most potential tyrosinase inhibitory activity among the extracts. Five main biocomponents were isolated and evaluated for their bioactivities. Among them, kaempferol (1) and quercetin (2) exhibited notable antioxidant activity in DPPH and ABTS assays. Particularly, kaempferol (1) performed the best α-glucosidase inhibitory effect, physcion (5) showed the strongest AChE inhibition, and quercetin (2) demonstrated the most potential for tyrosinase inhibitory activity. Further molecular docking studies revealed that there may be stronger binding mechanisms between bioactive components and target enzymes (including α-glucosidase, acetylcholinesterase, and tyrosinase) than the positive control. These findings suggest that bioactive extracts and compounds from the stems of R. formosana may have potential as natural antioxidant, anti-α-glucosidase, anti-AChE, and anti-tyrosinase drug candidates or dietary supplements for the management of oxidative stress-related conditions, including hyperglycemia, pigmentation disorders, and neurodegenerative diseases.

Antioxidants doi: 10.3390/antiox14010007

Authors: Yi-Chin Li Mei-Ling Cheng

Hypoxia/reoxygenation (HR) often occurs under cardiac pathological conditions, and HR-induced oxidative stress usually leads to cardiomyocyte damage. Carvedilol, a non-selective β-blocker, is used clinically to treat cardiac ischemia diseases. Moreover, Carvedilol has also been reported to have an antioxidant ability by reducing lipid peroxidation. However, the mechanism of Carvedilol to inhibit lipid peroxidation is still elusive. To explore the protective mechanism of Carvedilol to resist lipid peroxidation on cardiomyocytes, HL-1 cells were cultured under normoxia, hypoxia, and HR and treated with Carvedilol to investigate the alteration on metabolism, protein expression, and mRNA level to explain its oxidative mechanism. The study found that Carvedilol upregulated glutathione peroxidase 4 (GPX4) protein expression to resist HR-induced lipid peroxidation by metabolic remodeling under HR. Also, Carvedilol promoted ferroptosis-related genes, ferritin heavy chain 1 (FTH1) and ferritin light chain 1 (FTL1) mRNA levels, to reduce lipid peroxidation under both hypoxia and HR. In conclusion, our study explores a mechanism by which Carvedilol inhibits ferroptosis by upregulating GPX4, FTH1, and FTL1 levels to downregulate lipid peroxidation under HR. The study provides a potential strategy for using Carvedilol in clinical applications, inspiring further research and development in the area of heart diseases.

Antioxidants doi: 10.3390/antiox14010006

Authors: Marco Zeppieri Caterina Gagliano Fabiana D’Esposito Mutali Musa Irene Gattazzo Maria Sole Zanella Federico Bernardo Rossi Alessandro Galan Silvia Babighian

Omega-3 fatty acids are critical components of cell membranes, including those in the retina. Specifically, eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) are the primary omega-3 fatty acids that have been studied for their potential benefits in retinal health, preventing the progression of retinopathy. Several studies have shown that a higher intake of omega-3 fatty acids is associated with a lower risk of developing diabetic retinopathy and age-related macular degeneration (AMD). Reviewing clinical trials and observational studies that support the protective role of omega-3s in retinal disorders is essential. This comprehensive review aims to evaluate the current literature on the role of omega-3 fatty acids, exploring their mechanisms of action and anti-inflammatory, anti-angiogenic, and neuroprotective roles in the retina. Omega-3s have been shown to inhibit abnormal blood vessel growth in the retina, which is a significant factor in proliferative diabetic retinopathy and neovascular AMD. Furthermore, omega-3 fatty acids are often studied with other nutrients, such as lutein, zeaxanthin, and vitamins, for their synergistic effects on retinal health. Reviewing these combinations can help understand how omega-3s can be part of a comprehensive approach to preventing or treating retinopathies, especially in diabetic patients. This review emphasizes the preventive function of EPA and DHA in alleviating oxidative stress-related damage in retinal diseases, concentrating on their antioxidative mechanisms.

Antioxidants doi: 10.3390/antiox14010005

Authors: Ivette Martínez-Vieyra Isaac Hernández-Rojo Víctor Hugo Rosales-García Aracely Evangelina Chávez-Piña Doris Cerecedo

Oxidative stress is widely recognized as a key mechanism in the development of hypertension. Under pathological conditions, such as in hypertension, oxidative stress leads to irreversible posttranslational modifications of proteins, which result in loss of protein function and cellular damage. We have previously documented physiological and morphological changes across various blood and bone marrow cell lineages, all of which exhibit elevated oxidative stress. While cytoskeletal changes in erythrocytes have been well characterized in hereditary diseases, this is the first study, to our knowledge, to investigate cytoskeletal reorganization in erythrocytes from hypertensive patients. To this end, we compared the expression patterns and subcellular distribution of key cytoskeletal proteins in erythrocytes from hypertensive individuals with those from normotensive subjects using Western blot, flow cytometry, and confocal microscopy. Our results revealed the presence of three erythrocyte subpopulations with differential expression of glycophorin A. The persistent oxidative environment in hypertensive patients causes dysregulation in the expression of glycophorin A, Band 3 protein, protein 4.1, and ankyrin, as well as the reorganization of spectrin. These alterations in protein expression and distribution suggest that oxidative stress in hypertensive individuals may induce structural modifications, ultimately impairing erythrocyte membrane elasticity and function.

Antioxidants doi: 10.3390/antiox14010004

Authors: José A. Matamoros Sara Rubio-Casado José A. Fernández-Albarral Miguel A. Martínez-López Elena Salobrar-García Eva M. Marco Victor Paleo-García Rosa de Hoz Inés López-Cuenca Lorena Elvira-Hurtado Lidia Sánchez-Puebla José M. Ramírez Juan J. Salazar Meritxell López-Gallardo Ana I. Ramírez

Glaucoma is a neurodegenerative disease characterized by the loss of retinal ganglion cells (RGCs), with intraocular pressure (IOP) being its primary risk factor. Despite controlling IOP, the neurodegenerative process often continues. Therefore, substances with neuroprotective, antioxidant, and anti-inflammatory properties could protect against RGC death. This study investigated the neuroprotective effects on RGCs and visual pathway neurons of a compound consisting of citicoline and coenzyme Q10 (CoQ10) in a mouse model of unilateral, laser-induced ocular hypertension (OHT). Four groups of mice were used: vehicle group (n = 6), citicoline + CoQ10 group (n = 6), laser–vehicle group (n = 6), and laser–citicoline + CoQ10 group (n = 6). The citicoline + CoQ10 was administered orally once a day starting 15 days before laser treatment, continuing until sacrifice (7 days post-laser). Retinas, the dorsolateral geniculate nucleus (dLGN), the superior colliculus (SC), and the visual cortex (V1) were analyzed. The citicoline + CoQ10 compound used in the laser–citicoline + CoQ10 group demonstrated (1) an ocular hypotensive effect only at 24 h post-laser; (2) prevention of Brn3a+ RGC death in OHT eyes; and (3) no changes in NeuN+ neurons in the dLGN. This study demonstrates that the oral administration of the citicoline + CoQ10 combination may exert a neuroprotective effect against RGC death in an established rodent model of OHT.

Antioxidants doi: 10.3390/antiox14010003

Authors: Jeongmin Lee Seok Min Hong Yong Sung Choi Jinseok Lee Joon Hyung Yeo Sung Soo Kim Jae Min Lee Dong Keon Yon Seung Geun Yeo

Many studies have evaluated the roles of free radicals and reactive oxygen species (ROS) in various diseases. To date, however, no systematic review has specifically investigated the involvement of free radicals and ROS in acute otitis media (OM), OM with effusion, and chronic OM. The present study therefore assessed the roles of free radicals and ROS in OM. SCOPUS, PubMed, Cochrane Library, EMBASE, and Google Scholar were comprehensively searched to identify studies assessing the roles of free radicals and ROS in OM. Relevant studies were identified and their findings summarized. The literature search identified 21 studies. A review of their findings showed that the concentrations of hydrogen peroxide (H2O2), lipid hydroperoxide (LPO), and myeloperoxidase (MPO) were elevated in patients with acute OM; that the concentrations of H2O2, LPO, MPO, xanthine oxidase (XO), superoxide dismutase (SOD), glutathione peroxidase (GPX), malondialdehyde (MDA), hydroxyl radical (·OH), and nitrotyrosine were elevated in patients with OM with effusion; and that the levels of nitric oxide (NO), MPO, 4-hydroxynonenal (4-HNE), and malondialdehyde (MDA) were elevated in patients with chronic OM. Although some studies reported that SOD levels were increased in patients with OM with effusion, decreases in antioxidants such as SOD and catalase, as well as total antioxidant capacity, were observed across all types of OM. Although the levels of free radicals and ROS varied by type of OM, study design, control group selection, sample type, ROS and free radical markers, and antioxidant types, most studies showed increased ROS and decreased antioxidants in OM patients. These findings suggest that levels of free radicals and ROS are strongly associated with the pathophysiology of OM.

Antioxidants doi: 10.3390/antiox14010002

Authors: Ruizhi Hu Xizi Yang Jianhua He Shusong Wu

This review systematically examines the influence of oxidative stress on the reproductive function of male livestock, with a particular focus on the modulation of autophagy. Spermatogenesis, a highly precise biological process, is vulnerable to a range of internal and external factors, among which oxidative stress notably disrupts autophagic processes within the testes. This disruption results in diminished sperm quality, impaired testosterone synthesis, and compromised integrity of the blood–testis barrier. Furthermore, this review elucidates the molecular mechanisms by which oxidative stress-induced autophagy dysfunction impairs spermatogenesis and mitochondrial function, consequently reducing sperm motility. These findings aim to provide a theoretical foundation and serve as a reference for improving reproductive performance and sperm quality in livestock.

Antioxidants doi: 10.3390/antiox14010001

Authors: Daitoku Murakami Takahiro Fukazawa Michihito Kyo Mutsumi Miyauchi Shigehiro Ono Tomonao Aikawa Nobuyuki Hirohashi Keiji Tanimoto

Lactoferrin (LF) is an iron-binding glycoprotein of the transferrin family and has been suggested to have a variety of biological functions, including anticancer activity. However, the effects of LF and its mechanisms in anticancer therapies, especially in radiotherapy against cancer cells under hypoxic conditions, are not well-determined. In this study, we focused on the molecular mechanisms of LF functions in cells under hypoxic conditions. High-dose LF treatment showed cytotoxic activity in a variety of cells, including both non-cancer and cancer cells. Interestingly, hypoxic treatment increased the sensitivity to LF in some cancer cells but decreased it in non-cancer cells. LF treatment also altered sensitivity to radiation treatment: LF significantly increased the viability of irradiated KD non-cancer cells under hypoxic conditions but decreased that of HSC2 cancer cells. These effects were only observed when LF was treated within 3 h of irradiation, but not before irradiation. Importantly, knockdown of HIF1A counteracted these effects in both cell lines. Measurements of ROS activity showed that LF decreased ROS production in KD cells but increased it in HSC2 cells, resulting in a decrease in γH2AX foci in KD cells but an increase in HSC2 cells. RNA-seq and gene set enrichment analysis showed that LF treatment regulated gene expression related to the cell cycle, apoptosis, inflammation, and the NRF2 antioxidant signaling pathway. Quantitative RT-PCR confirmed the downregulation of the pro-apoptotic gene ASC in KD cells and the NRF2-regulated genes in HSC2 cells by LF treatment. Knockdown experiments confirmed the role of ASC in irradiated KD cells and NRF2 in irradiated HSC2 cells with LF treatment. In conclusion, lactoferrin was shown to affect radiation treatment by regulating apoptosis and NRF2 signaling in a cell type-specific manner under hypoxic conditions, suggesting its potential application as a protector or sensitizer for radiation therapy.

Antioxidants doi: 10.3390/antiox13121589

Authors: Maria Cidália R. Castro Pedro Veiga Rodrigues Vasco Cruz Ana Vera Machado

The packaging industry has made efforts to reduce food waste and improve the resilience of food systems worldwide. Active food packaging, which incorporates active agents, represents a dynamic area where industry and academia have developed new strategies to produce innovative and sustainable packaging solutions that are more compatible with conventional options. Due to health and environmental concerns, industries have sought alternatives to petroleum-based materials and have found biopolymers to be a viable option because of their biodegradable and safe nature. The combination of PLA/TPS has emerged as an effective system for packaging film; however, they are thermodynamically immiscible. This work highlights the development of a starch-based compatibilizer to connect the PLA and TPS phases by functionalizing maize starch with glycidyl methacrylate, glycerol, or garlic oil. Garlic oil was chosen for its plasticizing ability and antioxidant properties. The films produced exhibited excellent compatibility, with enhanced interfacial adhesion between PLA and TPS components. The introduction of compatibilizers also increased the systems’ crystallinity and improved their mechanical properties. The wettability of the films significantly increased with higher garlic oil content, along with enhanced antioxidant properties. These advancements will enable the production of a compatible PLA/TPS system with improved properties for application in the packaging industry.

Antioxidants doi: 10.3390/antiox13121588

Authors: Ahlam Harfoush Aseel Swaidan Salma Khazaal Elie Salem Sokhn Nabil Grimi Espérance Debs Nicolas Louka Nada El Darra

Tea, one of the most popular beverages worldwide, generates a substantial amount of spent leaves, often directly discarded although they may still contain valuable compounds. This study aims to optimize the extraction of polyphenols from spent black tea (SBT) and spent green tea (SGT) leaves while also exploring their antioxidant and antibacterial properties. Response surface methodology was utilized to determine the optimal experimental conditions for extracting polyphenols from SBT and SGT. The total phenolic content (TPC) was quantified using the Folin–Ciocalteu method, while antioxidant activity was evaluated through the DPPH assay. Antibacterial activity was assessed using the disk diffusion method. Additionally, high-performance liquid chromatography (HPLC) was employed to analyze the phytochemical profiles of the SBT and SGT extracts. Optimal extraction for SBT achieved 404 mg GAE/g DM TPC and 51.5% DPPH inhibition at 93.64 °C, 79.9 min, and 59.4% ethanol–water. For SGT, conditions of 93.63 °C, 81.7 min, and 53.2% ethanol–water yielded 452 mg GAE/g DM TPC and 78.3% DPPH inhibition. Both tea extracts exhibited antibacterial activity against Gram-positive bacteria, with SGT showing greater efficacy against S. aureus and slightly better inhibition of B. subtilis compared to SBT. No activity was observed against the Gram-negative bacteria E. coli and S. typhimurium. HPLC analysis revealed hydroxybenzoic acid as the main phenolic compound in SBT (360.7 mg/L), while rutin was predominant in SGT (42.73 mg/L). The optimized phenolic-rich extracts of SBT and SGT demonstrated promising antioxidant and antibacterial potential, making them strong candidates for use as natural health boosters in food products.

Antioxidants doi: 10.3390/antiox13121587

Authors: Cécile Méjécase Neelima Nair Hajrah Sarkar Pablo Soro-Barrio Maria Toms Sophia Halliday Katy Linkens Natalia Jaroszynska Constance Maurer Nicholas Owen Mariya Moosajee

Reactive oxygen species (ROS) within the retina play a key role in maintaining function and cell survival. However, excessive ROS can lead to oxidative stress, inducing dysregulation of metabolic and inflammatory pathways. The chmru848 zebrafish models choroideremia (CHM), an X-linked chorioretinal dystrophy, which predominantly affects the photoreceptors, retinal pigment epithelium (RPE), and choroid. In this study, we examined the transcriptomic signature of the chmru848 zebrafish retina to reveal the upregulation of cytokine pathways and glia migration, upregulation of oxidative, ER stress and apoptosis markers, and the dysregulation of glucose metabolism with the downregulation of glycolysis and the upregulation of the oxidative phase of the pentose phosphate pathway. Glucose uptake was impaired in the chmru848 retina using the 2-NBDG glucose uptake assay. Following the overexpression of human PFKM, partial rescue was seen with the preservation of photoreceptors and RPE and increased glucose uptake, but without modifying glycolysis and oxidative stress markers. Therapies targeting glucose metabolism in CHM may represent a potential remedial approach.

Antioxidants doi: 10.3390/antiox13121586

Authors: Vigneswari Sankara Narayanan Soven Dhawa Amritha Sukumaran Bharathi Hassan Ganesh Jeya Rajendran Kondapa Naidu Bobba Prasanna Ramani

A novel Schiff base ligand (L), bearing N2O2 donor sites, was derived from the condensation of 5-chloromethylisophthaldehyde and phenylpropanolamine (PPA). Mononuclear Co(II), Cu(II), and Zn(II) complexes were synthesized and were characterized by FTIR, UV–Vis, 1H NMR, ESI-mass spectroscopy, molar conductance, and thermal and electrochemical studies. The thermal investigation revealed that the complexes were stable up to 150–250 °C and began to degrade in stages, resulting in the development of respective metal oxides. The Coats–Redfern integration method was used to calculate the kinetic and thermodynamic parameters, the energy of activation (Ea), and changes in enthalpy (∆H), entropy (∆S), and free energy (∆G) for each step of the degradation processes. For stage I decomposition, the calculated activation energy values of the complexes follow the order of Ea [Cu(L)] > Ea [Co(L)(H2O)2] > Ea [Zn(L)]. The influence of the temperature on the efficacy of antioxidant activities of the complexes with DPPH assay, ABTS assay, and hydroxy radical assay was investigated at various concentrations using ascorbic acid (AA) as the reference. Antioxidant activity was assessed at multiple temperatures to ascertain whether these complexes may be applied in radiation therapy enhanced with hyperthermia and found to be stable. Subsequently, the Cu(II) complex (C2) demonstrated a greater cytotoxicity (IC50 = 5.16 µM) than Co(II), Zn(II), and conventional cisplatin when in vitro cytotoxicity was evaluated against the MCF-7 cell line using the MTT method. Analyses of the thermal stability and ROS scavenging ability of complexes have demonstrated that these complexes have potential in hyperthermic radiation therapy.

Antioxidants doi: 10.3390/antiox13121585

Authors: Tania Ciaglia Maria Rosaria Miranda Simone Di Micco Mariapia Vietri Gerardina Smaldone Simona Musella Veronica Di Sarno Giulia Auriemma Carla Sardo Ornella Moltedo Giacomo Pepe Giuseppe Bifulco Carmine Ostacolo Pietro Campiglia Michele Manfra Vincenzo Vestuto Alessia Bertamino

Based on the established neuroprotective properties of indole-based compounds and their significant potential as multi-targeted therapeutic agents, a series of synthetic indole–phenolic compounds was evaluated as multifunctional neuroprotectors. Each compound demonstrated metal-chelating properties, particularly in sequestering copper ions, with quantitative analysis revealing approximately 40% chelating activity across all the compounds. In cellular models, these hybrid compounds exhibited strong antioxidant and cytoprotective effects, countering reactive oxygen species (ROS) generated by the Aβ(25–35) peptide and its oxidative byproduct, hydrogen peroxide, as demonstrated by quantitative analysis showing on average a 25% increase in cell viability and a reduction in ROS levels to basal states. Further analysis using thioflavin T fluorescence assays, circular dichroism, and computational studies indicated that the synthesized derivatives effectively promoted the self-disaggregation of the Aβ(25–35) fragment. Taken together, these findings suggest a unique profile of neuroprotective actions for indole–phenolic derivatives, combining chelating, antioxidant, and anti-aggregation properties, which position them as promising compounds for the development of multifunctional agents in Alzheimer’s disease therapy. The methods used provide reliable in vitro data, although further in vivo validation and assessment of blood–brain barrier penetration are needed to confirm therapeutic efficacy and safety.

Antioxidants doi: 10.3390/antiox13121583

Authors: Beata Kruszewska-Naczk Mariusz Grinholc Aleksandra Rapacka-Zdonczyk

Antimicrobial blue light (aBL) has become a promising non-invasive method that uses visible light, typically within the 405–470 nm wavelength range, to efficiently inactivate a wide variety of pathogens. However, the mechanism of antimicrobial blue light (aBL) has not been fully understood. In this study, our research group investigated the sensitivity of Escherichia coli BW25113 single-gene deletion mutants to individual stressors generated by aBL. Sixty-four aBL-sensitive mutants were tested under conditions mimicking the stress generated by irradiation with aBL, with their growth defects compared to the wild-type strain. Results revealed no positive correlation between aBL and single stressors, indicating that aBL’s effectiveness is due to the simultaneous generation of multiple stressors. This multifactorial effect suggests that aBL targets microbial cells more precisely than single stressors such as hydrogen peroxide. No single gene knockout conferred specific resistance, highlighting aBL’s potential as an antimicrobial strategy.

Antioxidants doi: 10.3390/antiox13121584

Authors: Mikhail Yu. Artamonov Felix A. Pyatakovich Inessa A. Minenko

In regenerative medicine, mesenchymal stem cells (MSCs) have shown their importance and potential in tissue reconstruction and immune system modification. However, such cells’ potential is often diminished by factors such as oxidative stress, immune rejection, and inadequate engraftment. This review highlights the role of molecular hydrogen (H2) and cold atmospheric plasma (CAP) as adjunct therapies to improve the effectiveness of MSC therapy. H2 has strong antioxidative and anti-inflammatory actions as it quenches reactive oxygen species and positively stimulates the Nrf2 pathway that promotes MSC survival and life. CAP, being a modulated source of ROS and RNS, also assists MSCs by altering the cellular redox balance, thus facilitating cellular adaptation, migration, and differentiation. H2 and CAP in conjunction with each other assist in establishing an ambience favorable for promoting MSCs’ survival and growth abilities, and reduce the healing time in various pathways such as wound, neuroprotection, and ischemia. Besides these concerns, this review also covers the best administration routes and doses of H2 and CAP together with MSCs in therapy. This study informs on a novel dual method aimed at improving the outcome of MSC therapy while adding several molecular targets and relevant clinical uses concerning these therapies. Research of the future has to deal with bettering these protocols so that the therapeutic benefits can be maximized without long-term implications for clinical applications.

Antioxidants doi: 10.3390/antiox13121582

Authors: Maja Repajić Ivona Elez Garofulić Ena Cegledi Erika Dobroslavić Sandra Pedisić Ksenija Durgo Ana Huđek Turković Jasna Mrvčić Karla Hanousek Čiča Verica Dragović-Uzelac

To determine the optimal conditions of pressurized liquid extraction (PLE) and microwave-assisted extraction (MAE) of polyphenols from black chokeberry leaves (BCL), temperature, time and sample-to-solvent ratio (SSR) were varied to obtain maximum polyphenols yield. The extracts were analyzed for total polyphenols (TP) as well as individual ones (UPLC ESI MS2) and antioxidant capacity (FRAP, DPPH and ORAC). Moreover, the biological activity of the selected extracts was additionally determined. The optimal PLE and MAE conditions were 150 °C, 5 min extraction time and SSR 1:30 g/mL (TP 80.0 mg GAE/g dm), and 70 °C, extraction time 5 min and SSR 1:30 g/mL (TP 36.4 mg GAE/g dm), respectively. Both methods yielded similar polyphenol profiles (43 compounds) but differed quantitatively. MAE extracts contained more flavonols and phenolic acids, while PLE extracts had higher procyanidins and flavan-3-ols. Furthermore, the PLE extract exhibited a superior antioxidant capacity. This BCL extract also showed that it can protect against oxidative and DNA damage and can induce free radical formation and DNA damage, albeit at different doses. Moreover, it had a moderate antimicrobial activity against S. aureus and B. subtilis, while no antimicrobial activity was observed against Gram-negative bacteria as well as yeasts, lactic acid bacteria and molds.

Antioxidants doi: 10.3390/antiox13121581

Authors: Marcelo Villalobos-Pezos Ociel Muñoz-Fariña Kong Shun Ah-Hen María Fernanda Garrido-Figueroa Olga García-Figueroa Alexandra González-Esparza Luisbel González-Pérez de Medina José Miguel Bastías-Montes

Edible mushrooms have been part of the human diet for centuries. Traditionally, they have been used for culinary and medicinal purposes due to their chemical composition and nutritional value, including their high antioxidant activity attributed to key metabolites such as phenolic compounds and ergothioneine. P. ostreatus mushrooms, known for their potent antioxidant activity, are prone to spoilage shortly after harvest, making preservation methods necessary. Various drying methods were evaluated to determine their effects on physicochemical characteristics, antioxidant properties, and ergothioneine content. Mushrooms were subjected to freeze-drying (FD), hot-air-drying (HAD), and microwave-vacuum-drying (MVD). The rehydration rate, color, and microstructural characteristics of extracts from fresh and dried P. ostreatus mushrooms were evaluated. Additionally, the total soluble phenolic content and antioxidant activity were assessed using DPPH and ORAC assays, along with the determination of ergothioneine content. FD and HAD at 40 °C showed the best results regarding the physicochemical characteristics. In terms of total soluble phenolic content, antioxidant activity, and ergothioneine content, the mushrooms dried using HAD at 40 °C showed the best retention of bioactive compounds. Among the evaluated methods, HAD at 40 °C proved to be a suitable alternative for processing P. ostreatus.