Antioxidants

20 pages, 4732 KB

Open AccessArticle

Recovery of SIRT3-SOD2 Axis and Mitophagy by Short-Term Calorie Restriction in Old Rat Soleus Skeletal Muscle

by Rosa Di Lorenzo, Anna Picca, Guglielmina Chimienti, Christiaan Leeuwenburgh, Vito Pesce and Angela Maria Serena Lezza

Antioxidants 2025, 14(9), 1125; https://doi.org/10.3390/antiox14091125 - 17 Sep 2025

Abstract

Age-related mitochondrial dysfunction is involved in the progressive loss of mass and strength of skeletal muscle with aging. The effects of a short-term calorie restriction (ST-CR) were assessed in the oxidative skeletal soleus muscle (Sol) from 27-month-old rats and compared with those of [...] Read more.

Age-related mitochondrial dysfunction is involved in the progressive loss of mass and strength of skeletal muscle with aging. The effects of a short-term calorie restriction (ST-CR) were assessed in the oxidative skeletal soleus muscle (Sol) from 27-month-old rats and compared with those of a CR in combination with resveratrol (RSV) (ST-CR + RSV). PGC-1α and PRXIII proteins showed a marked decrease in both ST-CR and ST-CR + RSV rats. The SIRT3 protein presented a very relevant increase in both ST groups. ST-CR and ST-CR + RSV elicited a marked increase in SOD2 protein amount and activity. ST-CR and ST-CR + RSV led to recovery of the SIRT3-SOD2 axis as a fast/early response. ST-CR and ST-CR + RSV did not affect the MFN2 protein, whereas both treatments induced a relevant increase in DRP1 protein. ST-CR and ST-CR + RSV induced a decrease in Parkin protein, suggestive of rescued mitophagy, leading to the elimination of dysfunctional mitochondria. Such a response likely enhanced the fission-mediated elimination of mitochondria, supported by the marked increase in DRP1. MtDNA copy number and TFAM protein were not changed by any ST treatment. The mtDNA oxidative damage level was strongly increased by both ST treatments. All the effects elicited by ST-CR and ST-CR + RSV were specific to the oxidative type fibers. Full article

(This article belongs to the Special Issue Mitochondrial Oxidative Stress in Aging and Disease—2nd Edition)

34 pages, 1551 KB

Open AccessArticle

Brine Enriched with Olive Wastewater Phenols: A Green Strategy to Reduce Nitrites in Cooked Ham

by Dario Mercatante, Stefania Balzan, Sonia Esposto, Sara Barbieri, Federico Fontana, Luca Fasolato, Vincenzo De Rosa, Maurizio Servili, Agnese Taticchi, Enrico Novelli and Maria Teresa Rodriguez-Estrada

Antioxidants 2025, 14(9), 1124; https://doi.org/10.3390/antiox14091124 - 17 Sep 2025

Abstract

This study aimed to evaluate the effects of brine enriched with an olive vegetation water (OVW) extract on the physico-chemical, oxidative, and sensory characteristics of cooked ham during storage, as a strategy to partially or totally replace nitrites. Four brines formulated with different [...] Read more.

This study aimed to evaluate the effects of brine enriched with an olive vegetation water (OVW) extract on the physico-chemical, oxidative, and sensory characteristics of cooked ham during storage, as a strategy to partially or totally replace nitrites. Four brines formulated with different concentrations of nitrites in combination with 200 mg of OVW extract/kg product were tested; the cooked ham samples were sliced, placed in trays, packed in a protective atmosphere, and monitored for 30 days at 4 °C. The results showed that phenolic compounds derived from OVW effectively reduced lipid and protein oxidation, limiting the formation of secondary oxidation products such as thiobarbituric acid reactive substances, volatile aldehydes, and cholesterol oxides. Sensory analysis confirmed that the extract did not negatively affect the organoleptic properties of the ham, while also helping to preserve color stability. These findings suggest that brine enriched with OVW phenols can be a promising green strategy to reduce nitrites in cooked ham, which also promotes the sustainable valorization of olive oil by-products. Full article

(This article belongs to the Special Issue Agri-Food Wastes as Natural Source of Bioactive Antioxidants: Fourth Edition)

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23 pages, 2368 KB

Open AccessArticle

Depolymerization and Nanoliposomal Encapsulation of Grape Seed Condensed Tannins: Physicochemical Characterization, Stability, In Vitro Release and Bioaccessibility

by Carolina F. Morales, Marcela Zamorano, Natalia Brossard, Andreas Rosenkranz and Fernando A. Osorio

Antioxidants 2025, 14(9), 1123; https://doi.org/10.3390/antiox14091123 - 16 Sep 2025

Abstract

Condensed tannins from grape seed residues show high antioxidant activity but low oral bioavailability because of their high degree of polymerization and covalent interactions with proteins. This study aimed to improve their bioaccessibility through depolymerization and encapsulation. Depolymerization was carried out using microwave-assisted [...] Read more.

Condensed tannins from grape seed residues show high antioxidant activity but low oral bioavailability because of their high degree of polymerization and covalent interactions with proteins. This study aimed to improve their bioaccessibility through depolymerization and encapsulation. Depolymerization was carried out using microwave-assisted SN1 reactions with gallic acid as a nucleophile under food-grade conditions, mainly producing epicatechin monomers with 99.8% polymer degradation efficiency. Importantly, the inhibition of ABTS●+ and DPPH● radicals remained unaffected (p > 0.05), indicating that depolymerization preserved the antioxidants’ redox function, maintaining about 90% of their inhibition activity. The products were encapsulated in phosphatidylcholine liposomes, which had nanometric sizes and high encapsulation efficiency (83.11%), and remained stable for up to 60 days. In vitro release of nanoliposomal epicatechin in a D1 simulant was less than 10% after 48 h, fitting a Weibull model (β = 0.07), suggesting sub-diffusive transport and demonstrating high bioactive retention capacity in aqueous systems. During in vitro digestion, bioaccessibility of gallic acid and epicatechin reached 95.61 ± 0.58% and 98.56 ± 0.81%, respectively, with a 2333% increase in the bioaccessible mass of flavan-3-ols in native liposomal condensed tannins, which otherwise showed no detectable bioaccessibility. These findings highlight the potential of polyphenols from agro-industrial waste with enhanced bioaccessibility for applications in nutraceuticals and functional foods. Full article

(This article belongs to the Special Issue Antioxidants from Sustainable Food Sources)

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17 pages, 6618 KB

Open AccessArticle

Synechococcus sp. PCC 7002 Performs Anoxygenic Photosynthesis and Deploys Divergent Strategies to Cope with H₂S_n and H₂O₂

by Yafei Wang, Yue Meng, Hongwei Ren, Ranran Huang, Jihua Liu and Daixi Liu

Antioxidants 2025, 14(9), 1122; https://doi.org/10.3390/antiox14091122 - 16 Sep 2025

Abstract

Oxygenic and anoxygenic photosynthesis have long been considered defining traits of cyanobacteria. However, whether the important cyanobacterial genus Synechococcus is capable of anoxygenic photosynthesis remains unconfirmed. Here, we report that Synechococcus sp. PCC 7002 is capable of anoxygenic photosynthesis when sulfide (H₂ [...] Read more.

Oxygenic and anoxygenic photosynthesis have long been considered defining traits of cyanobacteria. However, whether the important cyanobacterial genus Synechococcus is capable of anoxygenic photosynthesis remains unconfirmed. Here, we report that Synechococcus sp. PCC 7002 is capable of anoxygenic photosynthesis when sulfide (H₂S) is supplied as the sole electron donor. Combining the targeted deletion of the sulfide: quinone oxidoreductase gene (Δsqr) with 3-(3,4-dichlorophenyl)-1,1-dimethylurea (DCMU) mediated the inhibition of photosystem II. We demonstrated that SQR-mediated H₂S oxidation sustains light-dependent CO₂ fixation in the absence of O₂ evolution. Our genome-wide transcriptomic profiling further revealed that polysulfide (H₂S_n) and hydrogen peroxide (H₂O₂) function as distinct signaling molecules in oxygenic and anoxygenic photosynthesis, modulating central carbon and energy metabolism. In central carbon metabolism, H₂S_n markedly upregulates the expression of key genes, including psbA, petC, rbcL, and rbcS, whereas H₂O₂ downregulates these genes. Within energy metabolism, both molecules converge on oxidative phosphorylation by upregulating genes encoding NADH dehydrogenase and ATP synthase. Furthermore, H₂Sₙ treatment uniquely induces sulfur-assimilation and ROS-detoxifying enzymes, conferring a markedly higher tolerance than H₂O₂. These findings provide direct evidence of anoxygenic photosynthesis in the genus Synechococcus and uncover a dual regulatory network that allows Synechococcus sp. PCC 7002 to balance redox homeostasis under fluctuating oxic/anoxic conditions. Full article

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58 pages, 1266 KB

Open AccessReview

Oxidative Stress and Antioxidants in Glioblastoma: Mechanisms of Action, Therapeutic Effects and Future Directions

by Agnieszka Nowacka, Maciej Śniegocki and Ewa Ziółkowska

Antioxidants 2025, 14(9), 1121; https://doi.org/10.3390/antiox14091121 - 15 Sep 2025

Abstract

Glioblastoma (GB) is an aggressive and treatment-resistant primary brain tumor with a dismal prognosis. Increasing evidence implicates oxidative stress as a central driver of its pathogenesis, progression, and resistance to therapy. The dynamic interplay between oxidative stress and antioxidant mechanisms is fundamental to [...] Read more.

Glioblastoma (GB) is an aggressive and treatment-resistant primary brain tumor with a dismal prognosis. Increasing evidence implicates oxidative stress as a central driver of its pathogenesis, progression, and resistance to therapy. The dynamic interplay between oxidative stress and antioxidant mechanisms is fundamental to understanding GBM biology and shaping novel therapeutic approaches. This review synthesizes current knowledge on the multifaceted role of redox biology in glioblastoma, highlighting the molecular mechanisms through which oxidative stress influences tumor proliferation, survival, immune evasion, and metabolic adaptation. Particular focus is given to the tumor microenvironment, hypoxia-driven reactive oxygen species, redox-regulating enzymes, and the immunosuppressive conditions fostered by oxidative stress. Antioxidants, in this context, demonstrate a dual role: while they can mitigate oxidative damage, their effects on cancer cells and treatment outcomes vary depending on the therapeutic setting. We further examine emerging strategies that target oxidative pathways, including small-molecule inhibitors, redox-modulating agents, and combinatorial approaches with standard treatments, while also addressing the complexities posed by antioxidant interventions. Preclinical and clinical findings are reviewed to underscore both the opportunities and challenges of exploiting redox vulnerabilities in GB. Ultimately, a deeper understanding of oxidative stress dynamics and antioxidant regulation may guide the development of innovative therapies that overcome resistance and improve outcomes for patients facing this devastating malignancy. Full article

(This article belongs to the Special Issue Antioxidant Phytochemicals for Promoting Human Health and Well-Being)

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19 pages, 2344 KB

Open AccessArticle

PTEN/PKM2/ERα-Driven Glyoxalase 1 Overexpression Sustains PC3 Prostate Cancer Cell Growth Through MG-H1/RAGE Pathway Desensitization Leading to H₂O₂-Dependent KRIT1 Downregulation

by Dominga Manfredelli, Camilla Torcoli, Marilena Pariano, Guido Bellezza, Tiziano Baroni, Vincenzo N. Talesa, Angelo Sidoni and Cinzia Antognelli

Antioxidants 2025, 14(9), 1120; https://doi.org/10.3390/antiox14091120 - 15 Sep 2025

Abstract

Glyoxalase 1 (Glo1) functions as a catalyst that neutralizes methylglyoxal (MG), a highly reactive glycating agent predominantly produced during glycolysis—a metabolic pathway upregulated in cancer cells. MG primarily reacts with the amino groups of proteins (especially at arginine residues), leading to the formation [...] Read more.

Glyoxalase 1 (Glo1) functions as a catalyst that neutralizes methylglyoxal (MG), a highly reactive glycating agent predominantly produced during glycolysis—a metabolic pathway upregulated in cancer cells. MG primarily reacts with the amino groups of proteins (especially at arginine residues), leading to the formation of a major advanced glycation end product known as MG-derived hydroimidazolone 1 (MG-H1). We previously demonstrated in PC3 human prostate cancer (PCa) cells that the PTEN/PKM2/ERα axis promotes their aggressive phenotype by regulating the Glo1/MG-H1 pathway. In this study, after confirming our earlier findings, we investigated the downstream mechanisms of the PTEN/PKM2/ERα/Glo1/MG-H1 axis in controlling PC3 cell growth, focusing on the role of RAGE, a high-affinity receptor for MG-H1; hydrogen peroxide (H₂O₂); and Krev interaction trapped 1 (KRIT1), an emerging tumor suppressor. Using genetic approaches and specific inhibitors/scavengers, we demonstrated that the PTEN/PKM2/ERα/Glo1/MG-H1 axis promotes PC3 cell growth—measured by proliferation and etoposide-induced apoptosis resistance—through a mechanism involving MG-H1/RAGE pathway desensitization that leads to H₂O₂-mediated KRIT1 downregulation. These findings support and expand the role of PTEN signaling in PCa progression and shed light on novel mechanistic pathways driven by MG-dependent glycative stress, involving KRIT1, in this still incurable stage of the disease. Full article

(This article belongs to the Special Issue Oxidative Stress in Metabolic Disorders and Non-Communicable Diseases (NCDs): Molecular Pathways and Genetic Influence)

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20 pages, 2613 KB

Open AccessArticle

Inhibitory Infrared Light Attenuates Mitochondrial Hyperactivity and Accelerates Restoration of Mitochondrial Homeostasis in an Oxygen–Glucose Deprivation/Reoxygenation Model

by Lucynda Pham, Tasnim Arroum, Paul T. Morse, Jamie Bell, Moh H. Malek, Thomas H. Sanderson and Maik Hüttemann

Antioxidants 2025, 14(9), 1119; https://doi.org/10.3390/antiox14091119 - 15 Sep 2025

Abstract

Ischemia/reperfusion (I/R) injury following stroke results in increased neuronal cell death due to mitochondrial hyperactivity. Ischemia results in loss of regulatory phosphorylations on cytochrome c oxidase (COX) and cytochrome c of the electron transport chain (ETC), priming COX for hyperactivity. During reperfusion, the [...] Read more.

Ischemia/reperfusion (I/R) injury following stroke results in increased neuronal cell death due to mitochondrial hyperactivity. Ischemia results in loss of regulatory phosphorylations on cytochrome c oxidase (COX) and cytochrome c of the electron transport chain (ETC), priming COX for hyperactivity. During reperfusion, the ETC operates at maximal speed, resulting in hyperpolarization of the mitochondrial membrane potential (ΔΨ_m) and reactive oxygen species (ROS) production. We have shown that COX-inhibitory near-infrared light (IRL) provides neuroprotection in small and large animal models of brain I/R injury. IRL therapy is non-invasive and non-pharmacological and does not rely on blood flow. We identified specific wavelengths of IRL, 750 and 950 nm, that inhibit COX activity. To model the mitochondrial effects following neuronal I/R, SH-SY5Y cells underwent oxygen–glucose deprivation/reoxygenation (OGD/R) ± IRL applied at the time of reoxygenation. Untreated cells exhibited ΔΨ_m hyperpolarization, whereas IRL treated cells showed no significant difference compared to control. IRL treatment suppressed ROS production, decreased the level of cell death, and reduced the time to normalize mitochondrial activity to baseline levels from 4–5 to 2.5 h of reperfusion time. We show that IRL treatment is protective by limiting ΔΨ_m hyperpolarization and ROS production, and by speeding up cellular recovery. Full article

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21 pages, 1447 KB

Open AccessArticle

Multielemental Profile for Seminal Plasma Through Inductively Coupled Plasma–Tandem Mass Spectrometry and Its Relationship with Seminal Parameters, Spermatic Biomarkers, and Oxidative Stress

by Andrea López-Botella, Natalia Cenitagoya-Alonso, Raquel Sánchez-Romero, Paula Sáez-Espinosa, Miranda Hernández-Falcó, María José Gómez-Torres and José Luis Todolí-Torró

Antioxidants 2025, 14(9), 1118; https://doi.org/10.3390/antiox14091118 - 15 Sep 2025

Abstract

The present study investigated the decline in human fertility by analyzing the multielemental profile of seminal plasma and its relationship with seminal parameters and sperm biomarkers. Twenty-nine donor seminal plasma samples were examined using inductively coupled plasma–tandem mass spectrometry (ICP-MS/MS). Method optimization demonstrated [...] Read more.

The present study investigated the decline in human fertility by analyzing the multielemental profile of seminal plasma and its relationship with seminal parameters and sperm biomarkers. Twenty-nine donor seminal plasma samples were examined using inductively coupled plasma–tandem mass spectrometry (ICP-MS/MS). Method optimization demonstrated that robust plasma conditions, including internal standardization and helium (He) collision gas, were essential to achieve reliable quantification. These conditions mitigated matrix effects and spectroscopic interferences, despite lower sensitivity. Elements such as copper (Cu), iron (Fe), manganese (Mn), strontium (Sr), titanium (Ti), vanadium (V), and chromium (Cr) were quantified, and several significant correlations were identified. Specifically, Cu was negatively correlated with seminal volume and positively correlated with sperm concentration and spontaneous acrosome reacted sperm, but negatively correlated with medium mitochondrial membrane potential (MMP); Mn showed negative associations with sperm vitality and medium MMP; Fe showed a negative correlation with motile sperm concentration (4 h); V was positively correlated with acrosome reacted sperm after acrosome reaction induction and with very low/medium MMP, whereas it was negatively associated with tyrosine phosphorylation; and Cr also showed a negative correlation with tyrosine phosphorylation. As, Mo, and Pb were detected in a few samples, limiting correlation analysis. From a functional perspective, elements such as As and Pb, as well as excess Cu or Fe, may contribute to oxidative stress by enhancing reactive oxygen species (ROS) generation and impairing antioxidant defenses. Conversely, essential metals, including Mn and Cu, at physiological concentrations act as cofactors of antioxidant enzymes and play a protective role against oxidative damage. Full article

(This article belongs to the Special Issue Oxidative and Nitrosative Stress in Male Reproduction)

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17 pages, 2585 KB

Open AccessArticle

Novel Hybrid Peptide DEFB126 (1-39)-TP5 Inhibits LPS-Induced Inflammatory Responses and Oxidative Stress by Neutralizing LPS and Blocking the TLR4/MD2-NFκB Signaling Axis

by Yuan Tang, Xuelian Zhao, Zetao Ding, Junyong Wang, Jing Zhang, Yichen Zhou, Marhaba Ahmat, Hao Wang, Yang Zhu, Baseer Ahmad, Zaheer Abbas, Dayong Si, Rijun Zhang and Xubiao Wei

Antioxidants 2025, 14(9), 1117; https://doi.org/10.3390/antiox14091117 - 14 Sep 2025

Abstract

Lipopolysaccharide (LPS), an essential structural molecule in the outer membrane of Gram-negative bacteria, is recognized as a principal trigger of inflammatory responses and oxidative stress. Thus, the control and clearance of LPS is essential to inhibit LPS-induced excessive inflammation, oxidative stress, and liver [...] Read more.

Lipopolysaccharide (LPS), an essential structural molecule in the outer membrane of Gram-negative bacteria, is recognized as a principal trigger of inflammatory responses and oxidative stress. Thus, the control and clearance of LPS is essential to inhibit LPS-induced excessive inflammation, oxidative stress, and liver injury. In recent years, some native bioactive peptides, such as human β-defensin 126 (DEFB126) and thymopentin (TP5), have been reported to have inhibitory effects against LPS-induced inflammation and oxidative stress. However, the cytotoxicity, weak stability, and poor biological activity have hindered their practical application and clinical development. The development of novel hybrid peptides is a promising approach for overcoming these problems. In this study, we designed a novel hybrid peptide [DTP, DEFB126 (1-39)-TP5] that combines the active center of DEFB126 and full-length thymopentin (TP5). Compared to the parental peptides, DTP has a longer half-life, lower cytotoxicity, and greater anti-inflammatory and antioxidant activity. The anti-inflammatory and antioxidant effects of DTP were demonstrated in a murine LPS-induced sepsis model, which showed that DTP successfully inhibited the indicators associated with LPS-induced liver injury; decreased the contents of TNF-α, IL-6, and IL-1β; increased the level of glutathione (GSH); and improved the activities of catalase (CAT) and superoxide dismutase (SOD). Furthermore, our study revealed that the anti-inflammatory and antioxidant activities of DTP were associated with LPS neutralization, blockade of LPS binding to the Toll-like receptor 4/myeloid differentiation factor 2 (TLR4/MD-2) complex, reduction in reactive oxygen species content, and inhibition of the activation of the nuclear factor kappa-B (NF-кB) signaling pathway. These results elucidate the structural and functional properties of the peptide DTP, reveal its underlying molecular mechanisms, and shed light on its potential as a multifunctional agent for applications in agriculture, food technology, and clinical therapeutics. Full article

(This article belongs to the Special Issue Antioxidant Peptides)

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25 pages, 7590 KB

Open AccessArticle

A Study of the Fruits of Catalpa bignonioides Walt.: Evaluation of the Antioxidant, Anti-Inflammatory, and Anti-Cancer Activities in Colorectal Adenocarcinoma Cells in Relation to Phytochemical Profile

by Clizia Bernardi, Thomas Gaslonde, Federica Finetti, Salim Benmaouche, Giulia Macrì, Annabelle Dugay, Claire Cuyamendous, Chouaha Bouzidi, Monica Rosa Loizzo, Philippe Belmont, Rosa Tundis, Lorenza Trabalzini and Brigitte Deguin

Antioxidants 2025, 14(9), 1116; https://doi.org/10.3390/antiox14091116 - 14 Sep 2025

Abstract

The chemical profiles and potential anti-inflammatory, antioxidant, and anticancer activities of the aqueous extract and fractions of fresh Catalpa bignonioides fruits were studied. Iridoids, flavonoids, and phenolic compounds represent the main phytochemical classes. Nine of the ten iridoids detected are acyl-iridoids. Significant amounts [...] Read more.

The chemical profiles and potential anti-inflammatory, antioxidant, and anticancer activities of the aqueous extract and fractions of fresh Catalpa bignonioides fruits were studied. Iridoids, flavonoids, and phenolic compounds represent the main phytochemical classes. Nine of the ten iridoids detected are acyl-iridoids. Significant amounts of catalpol and catalposide were found. The antioxidant activity of iridoids was demonstrated by HPTLC analysis coupled with a DPPH derivatization and by applying four in vitro tests, such as DPPH, ABTS, FRAP, and the β-carotene bleaching test. C. bignonioides extract and fractions were also evaluated for their anti-cancer activity using in vitro models of colorectal cancer (HT29 and HCT166 cell lines), and focusing on the effect of the different fractions on inflammation and oxidative stress, key factors that drive the onset and progression of colon cancer. Full article

(This article belongs to the Special Issue Antioxidant and Anti-Inflammatory Potential of Plants in Cancer Treatment)

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23 pages, 4376 KB

Open AccessArticle

Integrated Metabolomics and Transcriptomics Reveals Metabolic Pathway Changes in Common Carp Muscle Under Oxidative Stress

by Yongxiang Liu, Bing Li, Yiran Hou, Linjun Zhou, Qiqin Yang, Chengfeng Zhang, Hongwei Li, Jian Zhu and Rui Jia

Antioxidants 2025, 14(9), 1115; https://doi.org/10.3390/antiox14091115 - 14 Sep 2025

Abstract

Hydrogen peroxide (H₂O₂), a ubiquitous reactive oxygen species in aquatic ecosystems, has been shown to induce toxicological effects in aquatic animals. However, the molecular mechanisms underlying H₂O₂-mediated alterations in muscle quality and metabolic homeostasis remain [...] Read more.

Hydrogen peroxide (H₂O₂), a ubiquitous reactive oxygen species in aquatic ecosystems, has been shown to induce toxicological effects in aquatic animals. However, the molecular mechanisms underlying H₂O₂-mediated alterations in muscle quality and metabolic homeostasis remain largely unexplored. In this study, we performed integrated metabolomic and transcriptomic analyses to characterize the molecular mechanisms underlying H₂O₂-induced oxidative stress in fish muscle tissue. Common carp (Cyprinus carpio) were randomized into two groups: a control group (0.0 mM H₂O₂) and an H₂O₂-treated group (1.0 mM H₂O₂) for a 14-day exposure. Following the exposure, comprehensive analyses, including fatty acid composition, amino acid profiles, and multi-omics sequencing, were conducted to elucidate the metabolic responses to oxidative stress. The results showed neither the amino acid nor the fatty acid composition exhibited significant modifications following H₂O₂ exposure. Metabolomic profiling identified 83 upregulated and 89 downregulated metabolites, predominantly comprising organic acids and derivatives, lipids and lipid-like molecules. These differential metabolites were associated with histidine and purine-derived alkaloid biosynthesis, glyoxylate and dicarboxylate metabolism pathways. Transcriptomic analysis identified 470 upregulated and 451 downregulated differentially expressed genes (DEGs). GO enrichment analysis revealed that these DEGs were significantly enriched in muscle tissue development and transcriptional regulatory activity. KEGG analysis revealed significant enrichment in oxidative phosphorylation, adipocytokine signaling, and PPAR signaling pathways. The elevated oxidative phosphorylation activity and upregulated adipocytokine/PPAR signaling pathways collectively indicate H₂O₂-induced metabolic dysregulation in carp muscle. Through the integration of metabolomics and transcriptomics, this study offers novel insights into the toxicity of H₂O₂ in aquatic environments, elucidates adaptive mechanisms of farmed fish to oxidative stress, and provides a theoretical basis for developing antioxidant strategies. Full article

(This article belongs to the Special Issue Natural Antioxidants and Aquatic Animal Health—2nd Edition)

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31 pages, 9126 KB

Open AccessArticle

Integrated Phytochemical Profiling, GC-MS Characterization, and In Silico, In Vitro Evaluation of Synergistic Antimicrobial, Antioxidant, and Anti-Inflammatory Activities of Morus alba Bark and Pinus densiflora Extracts with Methyl Gallate

by Muhammad Aleem Abbas, Ga-Yeong Lee, Syed Al Jawad Sayem, Seung-Jin Lee and Seung-Chun Park

Antioxidants 2025, 14(9), 1114; https://doi.org/10.3390/antiox14091114 - 13 Sep 2025

Abstract

The growing challenge of antibiotic resistance and inflammation-related disorders calls for safe, multi-target therapeutic strategies. Morus alba (MOAL) and Pinus densiflora (PIDE) are known for their medicinal properties, yet their combined potential with methyl gallate (MG) has not been fully explored. In this [...] Read more.

The growing challenge of antibiotic resistance and inflammation-related disorders calls for safe, multi-target therapeutic strategies. Morus alba (MOAL) and Pinus densiflora (PIDE) are known for their medicinal properties, yet their combined potential with methyl gallate (MG) has not been fully explored. In this study, the phytochemical composition of MOAL and PIDE was characterized using GC–MS, and their combined antimicrobial, antioxidant, and anti-inflammatory activities were evaluated. Hydroethanolic extracts were prepared and assessed for antioxidant activity (DPPH assay), antibacterial activity (disk diffusion, MIC, time kill), and nitric oxide (NO) suppression in Lipopolysaccharide (LPS)-stimulated macrophages, alongside MTT cytotoxicity screening. MOAL exhibited a higher extraction efficiency, reaching 500 mg/mL at 4 h, whereas Pinus achieved 450 mg/mL at the same time point. Both exhibited a diverse and abundant phytochemical profile. The optimized blend (MOAL:PIDE:MG, 1:1:0.1) demonstrated significantly enhanced bioactivity, with over 90% DPPH scavenging with the low IC₅₀ value (66.62 mg/mL), potent inhibition of both Gram-positive and Gram-negative bacteria, and the strongest effect against Staphylococcus aureus (264 μg/mL). Time-kill assays confirmed rapid bactericidal action, and NO production was reduced by approximately 75% without cytotoxicity. Molecular docking identified a lead multi-target compound exhibiting strong binding affinities to COX-2, TNF-α, and Keap1, supporting its observed anti-inflammatory and antioxidant potential. These findings highlight the promise of synergistic phytochemical formulations as broad-spectrum, multifunctional therapeutic candidates, supporting further in vivo and clinical validation. Full article

(This article belongs to the Special Issue Bioactive Compounds: Antioxidant, Antibacterial, Anti-inflammatory Modulation)

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18 pages, 4016 KB

Open AccessArticle

The Supersulfide-Producing Activity of Rat Cystathionine γ-Lyase Is Irreversibly Inactivated by L-CysNO but Not by L-GSNO

by Shoma Araki, Tsuyoshi Takata, Sunghyeon Yoon, Shingo Kasamatsu, Hideshi Ihara, Hidehiko Nakagawa, Takaaki Akaike, Yukihiro Tsuchiya and Yasuo Watanabe

Antioxidants 2025, 14(9), 1113; https://doi.org/10.3390/antiox14091113 - 13 Sep 2025

Abstract

Cystathionine γ-lyase (CSE) is a pyridoxal 5′-phosphate (PLP)-dependent enzyme that catalyzes the final step of the transsulfuration pathway, converting cystathionine into cysteine. Additionally, CSE is also essential for the formation of cysteine hydropolysulfide (Cys-S-(S)n-H), known as supersulfides, by metabolizing cystine under pathological conditions. [...] Read more.

Cystathionine γ-lyase (CSE) is a pyridoxal 5′-phosphate (PLP)-dependent enzyme that catalyzes the final step of the transsulfuration pathway, converting cystathionine into cysteine. Additionally, CSE is also essential for the formation of cysteine hydropolysulfide (Cys-S-(S)n-H), known as supersulfides, by metabolizing cystine under pathological conditions. We previously reported that, during cystine metabolism, CSE undergoes self-inactivation through polysulfidation at the Cys136 residue. Here, contrary to the anticipated role of L-S-nitrosocysteine (L-CysNO) as a nitric oxide (NO) donor, we demonstrate that it serves as a substrate for CSE and that its metabolites inhibit the activity of the enzyme during L-CysNO metabolism. The in vitro incubation of CSE—but not the Cys136/171Val mutant—with L-CysNO resulted in the dose-dependent inhibition of supersulfide production, which was not reversed by the reducing agents. Notably, CSE activity remained unchanged upon preincubation with other NO donors, such as S-nitrosoglutathione or D-CysNO, but was inhibited when coincubated with cysteine. Furthermore, when PLP was removed from the CSE/L-CysNO premix, L-CysNO no longer inhibited CSE activity, suggesting that CSE metabolizes L-CysNO and that its metabolites contribute to enzyme inactivation. Indeed, we identified thionitrous acid and pyruvate as the primary CSE/L-CysNO reaction products. Thus, we establish L-CysNO as a CSE substrate and demonstrate that its metabolites act as enzyme inhibitors through a novel irreversible modification at the Cys136/171 residues. Full article

(This article belongs to the Special Issue Nitric Oxide (NO) and Hydrogen Sulfide (H₂S) in Biology, Illness, and Therapies—2nd Edition)

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20 pages, 5047 KB

Open AccessArticle

Physiological and Transcriptome Analyses Offer Insights into Revealing the Mechanisms of Red Tilapia (Oreochromis spp.) in Response to Carbonate Alkalinity Stress

by Wei Ye, Wen Wang, Jixiang Hua, Dongpo Xu and Jun Qiang

Antioxidants 2025, 14(9), 1112; https://doi.org/10.3390/antiox14091112 - 13 Sep 2025

Abstract

The utilization of saline–alkali water resources presents a promising approach for freshwater aquaculture. Red tilapia (Oreochromis spp.) exhibits moderate salinity tolerance, but its adaptation mechanism to alkaline conditions remains poorly understood. In the current study, five alkaline carbonate concentrations in a 60-day [...] Read more.

The utilization of saline–alkali water resources presents a promising approach for freshwater aquaculture. Red tilapia (Oreochromis spp.) exhibits moderate salinity tolerance, but its adaptation mechanism to alkaline conditions remains poorly understood. In the current study, five alkaline carbonate concentrations in a 60-day chronic stress experiment on red tilapia were evaluated. The experimental design included a control group (CA0, 0 mmol/L) and three treatment groups (CA10, 20 mmol/L; CA30, 30 mmol/L; and CA40 40 mmol/L). The results indicated that at alkaline carbonate concentrations exceeding 20 mmol/L, the gill filaments exhibited curling and deformation, the hepatocytes displayed migration, and tissue damage increased significantly. The gill’s antioxidant capacity initially decreased and then increased, with severe gill injury in the CA40 group, leading to significantly reduced levels of SOD, CAT, and GSH-PX compared to the CA40 group (p < 0.05). Conversely, the enzymatic activities related to energy metabolism showed an opposite trend under alkaline carbonate stress. The transcriptome analyses of gill tissues across five groups identified significant alterations in key pathways, including the metabolic process (endocytosis, focal adhesion, PI3K−Akt signaling pathway, MAPK signaling pathway, and Citrate cycle (TCA cycle)), and immune responses (mTOR signaling and NOD−like receptor signaling pathways). Additionally, we screened 13 differentially expressed genes (DEGs) as potential regulators of alkaline stress and validated their expression levels using quantitative real-time PCR (qPCR). This study preliminarily elucidated the molecular mechanism of red tilapia in the physiological regulation process under chronic alkaline stress, and offers a theoretical foundation for breeding programs aimed at developing alkali-tolerant strains for aquaculture in alkaline water environments. Full article

(This article belongs to the Section Health Outcomes of Antioxidants and Oxidative Stress)

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24 pages, 1355 KB

Open AccessReview

Carotenoids and Their Interaction with the Immune System

by Miguel Medina-García, Andrés Baeza-Morales, Pascual Martínez-Peinado, Sandra Pascual-García, Carolina Pujalte-Satorre, Rosa María Martínez-Espinosa and José Miguel Sempere-Ortells

Antioxidants 2025, 14(9), 1111; https://doi.org/10.3390/antiox14091111 - 12 Sep 2025

Abstract

Carotenoids are lipophilic pigments naturally occurring in plants and, to a lesser extent, in certain non-photosynthetic organisms. They play a critical role in human health due to their antioxidant and immunomodulatory properties. Key carotenoids such as β-carotene, lycopene, lutein, and zeaxanthin are capable [...] Read more.

Carotenoids are lipophilic pigments naturally occurring in plants and, to a lesser extent, in certain non-photosynthetic organisms. They play a critical role in human health due to their antioxidant and immunomodulatory properties. Key carotenoids such as β-carotene, lycopene, lutein, and zeaxanthin are capable of neutralizing reactive oxygen species, thereby mitigating oxidative stress—a major contributor to the onset and progression of chronic diseases. These compounds also modulate immune responses by influencing lymphocyte proliferation, enhancing natural killer cell activity, and regulating the production of pro- and anti-inflammatory cytokines. Such immunomodulatory effects are associated with a reduced risk of infectious diseases and have shown potential protective roles against inflammatory conditions, cardiovascular and neurodegenerative disorders, and certain types of cancer. Moreover, diets rich in carotenoids are linked to improved immune status, particularly in vulnerable populations such as the elderly and immunocompromised individuals. Despite strong epidemiological evidence, clinical trials involving carotenoid supplementation have produced mixed results, indicating that their effectiveness may depend on the broader dietary context and interactions with other nutrients. In summary, carotenoids are important dietary compounds that contribute to immune regulation and the prevention of various diseases, although further clinical research is needed to determine optimal intake levels and assess their full therapeutic potential. Full article

(This article belongs to the Special Issue Carotenoids in Health and Disease)

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40 pages, 2908 KB

Open AccessReview

Engineering Antioxidants with Pharmacological Applications: Biotechnological Perspectives

by Mădălina Paraschiv, Delia Turcov, Anca Zbranca-Toporaş, Bianca-Iulia Ciubotaru, Irina Grădinaru and Anca-Irina Galaction

Antioxidants 2025, 14(9), 1110; https://doi.org/10.3390/antiox14091110 - 12 Sep 2025

Abstract

Oxidative stress, a state resulting from an imbalance between the generation of reactive oxygen species (ROS) and the body’s antioxidant capacity, is a significant contributor to the development of various human pathologies, including malignancies, cardiovascular conditions, neurodegenerative disorders, and the aging process. Antioxidants, [...] Read more.

Oxidative stress, a state resulting from an imbalance between the generation of reactive oxygen species (ROS) and the body’s antioxidant capacity, is a significant contributor to the development of various human pathologies, including malignancies, cardiovascular conditions, neurodegenerative disorders, and the aging process. Antioxidants, both enzymatic and non-enzymatic, are vital in neutralizing free radicals and protecting against cellular damage. Given the limitations of synthetic antioxidants, such as potential toxicity and variable effectiveness, there has been a growing focus on biotechnological methods for producing these essential compounds. This review, titled “Engineering Antioxidants with Pharmacological Applications: Biotechnological Perspectives”, explores the latest developments in this field by examining how biological systems are being utilized to create a wide range of antioxidants. We discuss key production strategies, including the use of microbial cell factories, enzyme-driven synthesis, plant cell cultures, and metabolic engineering. The review provides specific examples of biotechnologically derived antioxidants, such as enzymatic defenses like superoxide dismutase, catalase, and glutathione peroxidase, as well as non-enzymatic molecules like carotenoids, polyphenols, and vitamins. We also evaluate the therapeutic potential of these bio-engineered antioxidants, analyzing preclinical and clinical data on their effectiveness in disease prevention and treatment. The mechanisms by which these compounds combat oxidative stress are also discussed. Finally, we address the current hurdles in scaling up production and managing costs while also outlining future research avenues, such as the creation of new production systems, advanced delivery technologies, and the discovery of novel antioxidant compounds through bioprospecting and synthetic biology. This comprehensive review highlights the potential of biotechnology to offer sustainable and impactful solutions for managing oxidative stress and enhancing overall health. Full article

(This article belongs to the Special Issue Multi-Target Profile of Antioxidant Compounds, Including Repurposing and Combination Strategies—2nd Edition)

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17 pages, 1651 KB

Open AccessArticle

The Role of Inulin in Maintaining Antioxidant Capacity and Enzymatic Activities of Jerusalem Artichoke (Helianthus tuberosus L.) Cultivars During Cold Storage

by Yuwen Mu, Bohua Zhang, Shiqi Lv, Fencan Li and Changming Zhao

Antioxidants 2025, 14(9), 1109; https://doi.org/10.3390/antiox14091109 - 12 Sep 2025

Abstract

Jerusalem artichoke (Helianthus tuberosus L.) is valued for its high inulin content and adaptability to marginal lands. This study investigated the changes in inulin content, antioxidant capacity, polyphenol concentrations, and enzymatic activities of eight cultivars during 60 days of cold storage. Inulin [...] Read more.

Jerusalem artichoke (Helianthus tuberosus L.) is valued for its high inulin content and adaptability to marginal lands. This study investigated the changes in inulin content, antioxidant capacity, polyphenol concentrations, and enzymatic activities of eight cultivars during 60 days of cold storage. Inulin levels ranged from 582.43 g/kg (LZJ006) to 809.70 g/kg (LZJ055), with LZJ047 maintaining the highest content throughout storage. The antioxidant potential, as measured by ferric reducing antioxidant power (FRAP) and 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay, declined across all cultivars, correlating with the reduction in inulin content. The polyphenol content varied significantly, with LZJ119 having 2.17 times more than LZJ010. POD activity increased, while catalase (CAT) and superoxide dismutase (SOD) activities fluctuated during the storage period. Hierarchical Cluster Analysis identified three distinct antioxidant clusters, revealing significant correlations between inulin content and key antioxidant parameters (CAT, FRAP, DPPH). These findings highlight the pivotal role of inulin in preserving the antioxidant system and bioactive properties of Jerusalem artichoke tubers during extended cold storage, providing valuable insights for post-harvest management and cultivar selection. Full article

(This article belongs to the Special Issue Food Proteins and Peptides as Antioxidants: Resource, Extraction, Functional Property and Application)

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24 pages, 11114 KB

Open AccessArticle

Deinoxanthin-Enriched Extracellular Vesicles from Deinococcus radiodurans Drive IL-10–Dependent Tolerogenic Programming of Dendritic Cells

by Jeong Moo Han, Jaeyoon Lim, Woo Sik Kim, Bo-Gyeong Yoo, Jong-Hyun Jung, Sangyong Lim and Eui-Baek Byun

Antioxidants 2025, 14(9), 1108; https://doi.org/10.3390/antiox14091108 - 12 Sep 2025

Abstract

Extracellular vesicles (EVs) derived from bacteria are emerging as potent bioactive carriers that affect host immunity. Deinococcus radiodurans, an extremophilic bacterium with strong antioxidant capacity, produces EVs enriched in deinoxanthin (DX), a carotenoid with a reactive oxygen species–scavenging activity. Here, we assessed [...] Read more.

Extracellular vesicles (EVs) derived from bacteria are emerging as potent bioactive carriers that affect host immunity. Deinococcus radiodurans, an extremophilic bacterium with strong antioxidant capacity, produces EVs enriched in deinoxanthin (DX), a carotenoid with a reactive oxygen species–scavenging activity. Here, we assessed the antioxidant activity of D. radiodurans-derived EVs (R1-EVs) in biochemical assays and their immunomodulatory effects on dendritic cells (DCs). R1-EVs exhibited significantly higher antioxidant activity than EVs from a DX-deficient mutant strain (ΔcrtI-EVs), consistent with DX enrichment. Bone marrow-derived DCs treated with R1-EVs in the presence of lipopolysaccharide displayed reduced expression of surface maturation markers and pro-inflammatory cytokines, while interleukin-10 (IL-10) production and antigen uptake were preserved, indicating a tolerogenic phenotype. This tolerogenic program led to decreased proliferation and cytokine production in allogeneic CD4⁺ and CD8⁺ T cells. Mechanistically, R1-EVs inhibited mitogen-activated protein kinase (MAPK) and nuclear factor kappa B (NF-κB) signaling pathways, key regulators of the DC activation. Importantly, IL-10 neutralization reversed these effects, restoring DC and T cell activation. Notably, ΔcrtI-EVs showed weaker antioxidant and immunoregulatory activities. Together, our findings identify R1-EVs as dual-functions, DX- and IL-10-dependent nanoplatform that integrates antioxidant and tolerogenic properties, with potential applications in inflammatory and autoimmune disease control. Full article

(This article belongs to the Special Issue Redox Regulation of Immune and Inflammatory Responses)

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17 pages, 1677 KB

Open AccessArticle

Association Between Blood Free Fatty Acid Concentrations in Midlife and Cerebral Small Vessel Disease

by Ryotaro Nukata, Yorito Hattori, Kotaro Noda, Takeshi Yoshimoto and Masafumi Ihara

Antioxidants 2025, 14(9), 1107; https://doi.org/10.3390/antiox14091107 - 12 Sep 2025

Abstract

Free fatty acids (FFAs) are a risk factor for recurrent ischemic stroke, primarily via the overproduction of reactive oxygen species. However, the association between FFA concentrations and cerebral small vessel disease (SVD), including lacunes, cerebral microbleeds, and white matter lesions on brain magnetic [...] Read more.

Free fatty acids (FFAs) are a risk factor for recurrent ischemic stroke, primarily via the overproduction of reactive oxygen species. However, the association between FFA concentrations and cerebral small vessel disease (SVD), including lacunes, cerebral microbleeds, and white matter lesions on brain magnetic resonance imaging, remains unclear. This study included 95 patients with acute ischemic stroke (median age: 59 [interquartile range: 49–73] years). The patients were divided into two groups: those aged ≤59 years (midlife patients) and those aged ≥60 years (late-life patients). In the midlife patients, the low serum total FFA concentration was an independent risk factor of lacunes (adjusted odds ratio [aOR]: 0.82, 95% confidence interval [CI]: 0.69–0.96; p = 0.013). Among FFA fractions, low serum free C14:0 (aOR: 0.80, 95% CI: 0.66–0.98; p = 0.028), and free C18:3n-3 (aOR: 0.93, 95% CI: 0.87–0.99; p = 0.015) concentrations were independent risk factors of lacunes in the midlife patients. However, the serum total FFA concentrations did not differ according to the SVD findings in the late-life patients. Therefore, low blood FFA concentrations in midlife can be a novel “nonvascular,” nonatheromatous risk factor of SVD, including the presence of lacunes identified on brain magnetic resonance imaging. Full article

(This article belongs to the Special Issue The Implication of Oxidative Stress on Human Aging and Associated Pathologies)

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