Antioxidants

23 pages, 6403 KB

Open AccessArticle

Dietary Thymol–Carvacrol Cocrystal Supplementation Improves Growth Performance, Antioxidant Status, and Intestinal Health in Broiler Chickens

by Jingzhe Yang, Changjin Li, Shuzhen Jiang, Yuemeng Fu, Guohui Zhou, Yufei Gao, Weiren Yang and Yang Li

Antioxidants 2025, 14(11), 1323; https://doi.org/10.3390/antiox14111323 (registering DOI) - 1 Nov 2025

Abstract

This study investigated the impacts of dietary thymol–carvacrol cocrystal (CEO) supplementation on broiler production performance, antioxidant status, intestinal health, and cecal microbiota. Eight hundred one-day-old chicks were randomly divided into four groups, receiving basal diets supplemented with 0, 40, 60, or 80 mg/kg [...] Read more.

This study investigated the impacts of dietary thymol–carvacrol cocrystal (CEO) supplementation on broiler production performance, antioxidant status, intestinal health, and cecal microbiota. Eight hundred one-day-old chicks were randomly divided into four groups, receiving basal diets supplemented with 0, 40, 60, or 80 mg/kg CEO. The results showed that CEO addition increased average daily gain, superoxide dismutase activity in the serum, liver, and jejunum, jejunal villus height/crypt depth ratio, cecal butyric acid concentration, and Lactobacillus abundance, while reducing serum alanine transaminase activity and malondialdehyde content in the serum, liver, and jejunum. Furthermore, 60 mg/kg CEO enhanced the final body weight, dressing percentage, serum total protein and glucose levels, and jejunal trypsin and amylase activities, while lowering the feed-to-gain ratio and serum cholesterol, urea nitrogen, and aspartate transaminase concentrations; it also increased the activities of superoxide dismutase, catalase, and glutathione and mRNA expressions of related genes in the liver and jejunum. It also increased cecal concentrations of acetic acid and isovalerate acid, while decreasing serum diamine oxidase and D-lactate concentrations, as well as malondialdehyde concentrations in the serum, liver, and jejunum. Therefore, dietary CEO supplementation improved the production performance, antioxidant status, and liver and gut health and function in broilers, with 60 mg/kg CEO demonstrating the most pronounced effects. Full article

(This article belongs to the Special Issue Oxidative Stress in Animal Reproduction and Nutrition)

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26 pages, 2614 KB

Open AccessArticle

Melatonin Improves Bovine Embryo Production and Quality via Antioxidant, Metabolic, and Epigenetic Pathways

by Hallya Beatriz Sousa Amaral, Márcia Marques Silveira, Ana Caroline Chaves Vall Nicolás, Laryssa Ketelyn Lima Pimenta, José Eduardo Vieira Chaves, Alexandre Rodrigues Caetano, Maurício Machaim Franco and Margot Alves Nunes Dode

Antioxidants 2025, 14(11), 1322; https://doi.org/10.3390/antiox14111322 (registering DOI) - 1 Nov 2025

Abstract

This study aimed to evaluate the effects of melatonin supplementation during bovine in vitro embryo production (IVEP) on embryonic development and quality, oxidative stress, lipid metabolism, mitochondrial activity, gene expression, DNA methylation patterns, and cryotolerance. Four treatments were tested: control (without melatonin), melatonin [...] Read more.

This study aimed to evaluate the effects of melatonin supplementation during bovine in vitro embryo production (IVEP) on embryonic development and quality, oxidative stress, lipid metabolism, mitochondrial activity, gene expression, DNA methylation patterns, and cryotolerance. Four treatments were tested: control (without melatonin), melatonin at maturation (IVM + Mlt), culture (IVC + Mlt), and both treatments (IMV/IVC + Mlt). Melatonin significantly improved blastocyst rate and developmental kinetics on D7, reduced ROS and intracellular lipid levels, and increased mitochondrial activity. The most significant effects were observed in the IVC + Mlt group. Melatonin modulated antioxidant (SOD1, Cat, and GSS) and epigenetic (TET1, TET3, and DNMT3A) genes, and although it did not alter lipid gene expression, it reduced lipid content. Methylation analysis showed hypomethylation patterns in repetitive regions (Satellite I and LINE-1), which were even more pronounced in the melatonin-treated groups. However, no significant differences were observed between treatments in terms of cryotolerance or apoptosis rates. These findings suggest that melatonin exerts positive multifactorial effects, regardless of the supplementation stage. In particular, its addition during the IVC phase appears to provide greater benefits to embryos by improving their quality. Full article

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23 pages, 5086 KB

Open AccessArticle

PPARα-Mediated Fatty Acid Catabolism in Astrocytes Was Involved in Improvement of Cognitive Dysfunction by Phlorizin in APP/PS1 Mice

by Yan Fu, Xuya Zhang, Lingling Li, Hong Jiang, Qiaozhi Ren, Tianxing Yi, Yali Zhang and Yi Lu

Antioxidants 2025, 14(11), 1321; https://doi.org/10.3390/antiox14111321 (registering DOI) - 31 Oct 2025

Abstract

Central lipid metabolism disorders are crucial for the development of Alzheimer’s disease (AD). Phlorizin (PHZ) improved lipid metabolism abnormalities in AD nematodes, but its mechanism of action in improving AD-related symptoms and whether it can alleviate AD cognitive impairment remain unclear. To elucidate [...] Read more.

Central lipid metabolism disorders are crucial for the development of Alzheimer’s disease (AD). Phlorizin (PHZ) improved lipid metabolism abnormalities in AD nematodes, but its mechanism of action in improving AD-related symptoms and whether it can alleviate AD cognitive impairment remain unclear. To elucidate the effects and mechanisms of PHZ on lipid metabolism disorders in an AD model, gavage administration of PHZ for 8 weeks improved cognitive dysfunction and lipid disorders in APPswe/PSEN1dE9 (APP/PS1) mice. Concurrently, in astrocytes induced by palmitic acid (PA)- mediated lipid metabolic disorder, PHZ treatment improved astrocytic lipid accumulation by upregulating the target peroxisome proliferator-activated receptor α (PPARα) and its downstream pathways, thereby promoting astrocytic fatty acid oxidation. We validated PHZ’s strong in vitro binding affinity with PPARα. Co-culture systems of lipid-metabolically disordered astrocytes and neurons further demonstrated that PHZ significantly improved neuronal cell viability and reduced intracellular lipid accumulation, thereby decreasing the expression of enzymes associated with β-amyloid protein (Aβ) production. This study demonstrates that gavage administration of PHZ for 2 months improves cognitive deficits and pathological markers in AD mice. Furthermore, at the cellular level, PHZ may exert its effects by enhancing astrocytic lipid metabolism, thereby preventing neuronal lipotoxicity and mitigating AD progression. Full article

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

21 pages, 3534 KB

Open AccessArticle

Chamazulene Induces Metabolic Reprogramming and Mitigates Inflammation in Photoaged Skin: PPARα/γ as Potential Regulators

by Ying Zhou, Wencui Wang, Lei He, Nan Zhang, Bowen Zhou, Zimeng Chen, Li Ma and Lei Yao

Antioxidants 2025, 14(11), 1320; https://doi.org/10.3390/antiox14111320 (registering DOI) - 31 Oct 2025

Abstract

Chamazulene (CHA) is a brilliant blue compound present in Artemisia sieversiana Ehrhart ex Willd. essential oil (AEO). We have previously reported that both CHA and AEO can shield the skin from UVB damage, exhibiting significant anti-photoaging effects. However, the molecular mechanisms underlying CHA’s [...] Read more.

Chamazulene (CHA) is a brilliant blue compound present in Artemisia sieversiana Ehrhart ex Willd. essential oil (AEO). We have previously reported that both CHA and AEO can shield the skin from UVB damage, exhibiting significant anti-photoaging effects. However, the molecular mechanisms underlying CHA’s photoprotective properties are still unclear. Herein, we integrated transcriptomics, targeted fatty acid profile, and untargeted metabolomics analyses on the dorsal skin of mice exposed to UVB with or without 0.4% CHA topical treatment. The results showed that CHA upregulated key genes involved in fatty acid metabolism, including two peroxisome proliferator-activated receptor (PPAR) subtypes, i.e., PPARα and PPARγ, in mouse skin. The CHA treatment elevated levels of various saturated, monounsaturated, and polyunsaturated fatty acids, and it especially restored n-3/n-6 polyunsaturated fatty acid homeostasis and downregulated the p38 MAPK/COX-2 pathway. Additionally, CHA enhanced skin non-essential amino acid metabolism, likely via PPARα. In conclusion, our study indicates that CHA may mitigate UVB-induced photoaging by inducing metabolic reprogramming and suppressing inflammation, and the findings suggest that the activation of PPARα/γ may play a vital role in these observed effects, thereby establishing CHA as a promising topical agent against UVB-induced photoaging. Full article

(This article belongs to the Special Issue Natural Antioxidants in Pharmaceuticals and Dermatocosmetology)

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15 pages, 810 KB

Open AccessArticle

Extraction, Phytochemical Analysis, and Bioactivity Evaluation of Polyphenols from Kunzea ericoides (Kanuka) Plant

by Harmandeep Dhaliwal, Yan Li and Michelle Yoo

Antioxidants 2025, 14(11), 1319; https://doi.org/10.3390/antiox14111319 (registering DOI) - 31 Oct 2025

Abstract

Kunzea ericoides (kanuka), a native plant of New Zealand, has a significant role in traditional medicine due to the presence of essential oils. Apart from these oils, this plant also is a source of many bioactive compounds, majority of which are polyphenols. However, [...] Read more.

Kunzea ericoides (kanuka), a native plant of New Zealand, has a significant role in traditional medicine due to the presence of essential oils. Apart from these oils, this plant also is a source of many bioactive compounds, majority of which are polyphenols. However, there is lack of sufficient data supporting the extraction of polyphenols from kanuka plant leaves and investigating its bioactivity and phytochemical properties. The study aims to extract polyphenols from kanuka plant leaves with a conventional solvent-based method and determine the phytochemical analysis as well as bioactive potential. Extraction was performed with methanol and acetone as solvents. Polyphenolic prolife was analyzed with LC-MS. Bioactive analysis of kanuka leaf extract was carried out to determine total phenolic content and antioxidant activity. We investigated the cytotoxic effect of kanuka leaf extract on two triple-negative breast cancer cells—MDA-MB-231 and BT-549. LC-MS analysis confirmed kanuka leaf extract is a source of many polyphenols, some giving very prominent signals on TIC scan. Ten polyphenolic compounds were confirmed to be present in kanuka leaf extract based on MRM analysis. FRAP-CUPRAC analysis indicated significant antioxidant activity in the kanuka leaf extract. Antiproliferative analysis has confirmed cytotoxicity of the kanuka leaf extract on the triple-negative breast cancer cell lines. This study indicates that Kunzea ericoides leaf extract, rich in polyphenols, shows promising antioxidant and antiproliferative potential, warranting further investigation for therapeutic applications. Full article

(This article belongs to the Special Issue Phytochemical Profiling, Antioxidant Activity and Therapeutic Properties of Medicinal Plants)

20 pages, 2135 KB

Open AccessArticle

Xingkai Lake Topmouth Culter (Culter alburnus) Exhibits Biochemical and Histopathological Alterations upon Acute Ammonia Exposure

by Junfei Yu, Hongling Yang, Guohe Cai, Jianming Xu, Banghua Xia and Yunzhang Sun

Antioxidants 2025, 14(11), 1318; https://doi.org/10.3390/antiox14111318 (registering DOI) - 31 Oct 2025

Abstract

The Xingkai Lake topmouth culter (Culter alburnus) is an endemic, economically valuable fish in Heilongjiang that is highly sensitive to ammonia. However, the systemic effects of acute ammonia stress on its liver have not been determined. The objective of this study [...] Read more.

The Xingkai Lake topmouth culter (Culter alburnus) is an endemic, economically valuable fish in Heilongjiang that is highly sensitive to ammonia. However, the systemic effects of acute ammonia stress on its liver have not been determined. The objective of this study was to elucidate the changes in and relationships among stress biomarkers, antioxidant defense mechanisms, apoptosis indicators, and histopathological alterations in the liver of C. alburnus, a fish species native to Xingkai Lake, China, under acute ammonia exposure. Guided by the findings of a 96 h-LC₅₀ assay, the researchers subjected the fish to 48 h of acute exposure at specified total ammonia nitrogen (TAN) concentrations of 30 mg/L, 36 mg/L, and 40 mg/L. A comprehensive assessment of physiological and biochemical markers, including cortisol (COR), blood ammonia (Amm), blood glucose (Glu), aspartate aminotransaminase (AST), alanine aminotransaminase (ALT), catalase (CAT), superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), and malondialdehyde (MDA), revealed pronounced physiological stress and oxidative damage, particularly in the high-concentration groups. The physiological effects of ammonia exposure on C. alburnus showed a clear concentration and time dependence. Notably, elevated ammonia levels significantly upregulated apoptosis-associated genes such as P53, Bax, and Caspase-3. These findings were further substantiated by terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assays and histopathological examinations. Overall, the study demonstrated that acute ammonia exposure exerted substantial impacts on the physiological, biochemical, and genetic expression profiles of C. alburnus in Xingkai Lake, leading to sustained stress and oxidative damage, especially at elevated concentrations (30–40 mg/L). Full article

(This article belongs to the Special Issue Reactive Oxygen Species Signalling and Oxidative Stress in Fish)

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19 pages, 4589 KB

Open AccessArticle

Digital Microfluidics-Driven Cell-Free Protein Synthesis Platform Reveals Expression and Stability Determinants for Phytoglobins and Cysteine-to-Alanine Substituted Variants

by Leonard Groth and Leif Bülow

Antioxidants 2025, 14(11), 1317; https://doi.org/10.3390/antiox14111317 (registering DOI) - 31 Oct 2025

Abstract

Heme proteins are central to metabolism and stress responses but remain challenging to express recombinantly due to cytotoxicity and folding constraints. Phytoglobins (Pgbs) exemplify these difficulties, as expression protocols often fail to translate across protein species. Here, we used a cell-free protein synthesis [...] Read more.

Heme proteins are central to metabolism and stress responses but remain challenging to express recombinantly due to cytotoxicity and folding constraints. Phytoglobins (Pgbs) exemplify these difficulties, as expression protocols often fail to translate across protein species. Here, we used a cell-free protein synthesis (CFPS) platform powered by digital microfluidics to screen expression determinants for sugar beet Pgb 1.2 (BvPgb 1.2), its C86A variant, and three of eight newly identified oat Pgbs (AsPgbs), including their cysteine-to-alanine substituted variants. Benchmarking with multiple solubility tags and cell-free blends revealed protein- and variant-specific preferences, with alanine substitutions frequently improving expression and purification yields. Oxidative additives such as glutathione disulfide, alone or combined with protein disulfide isomerase, consistently enhanced production, underscoring the importance of redox environments for Pgb stability. Two selected variants were scaled up and yielded putative soluble apo-form proteins. The results highlight how CFPS enables rapid, parallelized identification of expression requirements while uncovering the role of conserved cysteines and redox conditions in Pgb biogenesis. Full article

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

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15 pages, 1210 KB

Open AccessArticle

Tobacco Smoke Exposure and Oxidative Stress: The Role of Circulating Lipopolysaccharides in Heated and Conventional Products

by Lorenzo Loffredo, Enrico Maggio, Simona Bartimoccia, Arianna Magna, Chiara Maria Totè, Chiara Bagnato, Bianca Laura Cinicola, Federica Armeli, Angela Leonardo, Alessandra D’Amico, Ernesto Greco, Giacomo Frati, Giuseppe Biondi-Zoccai, Alberto Spalice, Antonio Angeloni, Pasquale Pignatelli, Francesco Violi, Anna Maria Zicari, Roberto Carnevale and Smoking Prevention Study Group

Antioxidants 2025, 14(11), 1316; https://doi.org/10.3390/antiox14111316 (registering DOI) - 31 Oct 2025

Abstract

Background: Exposure to tobacco smoke, from conventional tobacco cigarettes (CTC) or heated tobacco products (HTPs), increases oxidative stress, causing endothelial dysfunction and higher cardiovascular risk. It is unclear whether smoke exposure also promotes low-grade endotoxemia, potentially activating NADPH oxidase and further impairing endothelial [...] Read more.

Background: Exposure to tobacco smoke, from conventional tobacco cigarettes (CTC) or heated tobacco products (HTPs), increases oxidative stress, causing endothelial dysfunction and higher cardiovascular risk. It is unclear whether smoke exposure also promotes low-grade endotoxemia, potentially activating NADPH oxidase and further impairing endothelial function. This study assessed serum lipopolysaccharide (LPS) levels in children and adults actively or passively exposed to conventional cigarette smoke or HTPs, compared with non-exposed controls. Methods: We conducted a cross-sectional study comprising 26 children passively exposed to HTPs, 26 children exposed to CTC, and 26 unexposed controls, as well as 20 adult chronic HTP users, 20 chronic CTC, and 20 non-smoking adults. Circulating LPS was measured alongside oxidative stress markers (NOX2, H₂O₂), endothelial function, intestinal permeability (zonulin), and nicotine exposure (serum cotinine). Results: Exposed children had higher cotinine, LPS, and zonulin than controls, with no differences between HTP and CTC groups. Multiple linear regression analysis identified cotinine (β = 0.343; p = 0.005) and zonulin (β = 0.441; p < 0.001) as independent LPS predictors. In adults, LPS and zonulin were higher in both smoker groups versus controls; zonulin (β = 0.477; p < 0.001) and nitric oxide bioavailability (β = −0.307; p = 0.007) independently predicted LPS. Conclusions: Passive and active exposure to CTC or HTPs increases low-grade endotoxemia and zonulin, potentially driving NOX2-mediated oxidative stress. Full article

(This article belongs to the Special Issue Cigarette Smoke and Oxidative Stress)

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26 pages, 12307 KB

Open AccessArticle

Parthenolide Restores Testosterone Biosynthesis After Nanoplastic Exposure by Blocking ROS-Driven NF-κB Nuclear Translocation

by Peng Zhao, Hao Yan, Runchang Wang, Jie Zhao, Xiangqin Zheng, Dinggang Li, Xitong Guo, Fengming Ji, Chunlan Long, Lianju Shen, Guanghui Wei and Shengde Wu

Antioxidants 2025, 14(11), 1315; https://doi.org/10.3390/antiox14111315 (registering DOI) - 31 Oct 2025

Abstract

Nanoplastics are pervasive contaminants that adversely affect male reproductive function, yet the molecular basis of polystyrene nanoplastic (PS-NP) toxicity in immature testes and effective preventive strategies remain unclear. Here, male mice (postnatal days 22–35, PND 22–35) and TM3 Leydig cells were exposed to [...] Read more.

Nanoplastics are pervasive contaminants that adversely affect male reproductive function, yet the molecular basis of polystyrene nanoplastic (PS-NP) toxicity in immature testes and effective preventive strategies remain unclear. Here, male mice (postnatal days 22–35, PND 22–35) and TM3 Leydig cells were exposed to graded PS-NPs, followed by transcriptomic profiling to identify differentially expressed genes (DEGs). Candidate therapeutics were prioritized using Connectivity Map (CMap) analysis and molecular docking, and protein interactions were examined by co-immunoprecipitation (Co-IP). PS-NPs accumulated in immature testes, eliciting excessive reactive oxygen species (ROS) and activation of NF-κB. These events coincided with the downregulation of steroidogenic enzymes (CYP11A1 and StAR) and disruption of testicular microarchitecture. In TM3 cells, PS-NPs suppressed testosterone synthesis in a concentration-dependent manner; this effect was fully reversed by pretreatment with N-acetylcysteine (NAC) or Bay 11-7082. Co-IP demonstrated p65–steroidogenic factor-1 (SF-1) binding consistent with formation of a transcriptional repressor complex targeting steroidogenic genes. CMap and docking analyses nominated parthenolide (PTL) as a candidate inhibitor of NF-κB nuclear translocation (predicted binding affinity, −6.585 kcal/mol), and PTL mitigated PS-NP-induced impairment of testosterone synthesis in vitro. Collectively, these data indicate that PS-NPs disrupt testosterone biosynthesis in immature testes through the ROS/NF-κB/p65–SF-1 axis, while PTL emerges as a candidate small molecule to counter nanoplastic-associated reproductive toxicity. These findings underscore translational relevance and support future evaluation under chronic low-dose exposure conditions, including in vivo validation of PTL efficacy, pharmacokinetics, and safety. Full article

(This article belongs to the Special Issue Oxidative Stress Induced by Micro(Nano)plastics)

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25 pages, 10151 KB

Open AccessArticle

Extraction, Purification and Identification of Bovine Lung Peptides and Its Antioxidant Effects on H₂O₂-Induced HepG2 Cells and Mice with Alcoholic Liver Injury

by Xingyu Xiao, Xunming Zhang, Yi Li, Tong Su, Shuo Zheng, Jiayuan Fang, Qinchuan Lv, Dacheng Wang and Linlin Hao

Antioxidants 2025, 14(11), 1314; https://doi.org/10.3390/antiox14111314 (registering DOI) - 31 Oct 2025

Abstract

In this study, we constructed an extraction process for bovine lung peptide-1 (BLP-1) derived from bovine lung tissue utilizing single-factor optimization combined with response surface methodology. We systematically analyzed its antioxidant activity, biological safety, and therapeutic mechanisms against alcoholic liver disease (ALD). In [...] Read more.

In this study, we constructed an extraction process for bovine lung peptide-1 (BLP-1) derived from bovine lung tissue utilizing single-factor optimization combined with response surface methodology. We systematically analyzed its antioxidant activity, biological safety, and therapeutic mechanisms against alcoholic liver disease (ALD). In vitro experiments demonstrated that BLP-1 exhibits excellent scavenging activity against various free radicals, while exhibiting no significant cytotoxicity or hemolytic activity. In a model of H₂O₂-induced oxidative damage in HepG2 cells, BLP-1 significantly alleviated oxidative stress injury by upregulating the activities of intracellular antioxidant enzymes. Animal experiments further confirmed that BLP-1 significantly reduced serum levels of transaminase, inhibited the release of inflammatory factors, enhanced antioxidant enzyme activity, and ameliorated lipid peroxidation and pathological injury in ALD mice. By combining liquid chromatography-tandem mass spectrometry (LC-MS/MS) with bioinformatics, we screened 12 novel antioxidant peptides. Among these, the binding energies of GP9, FG6, and WG6 to Keap1 were −10.2, −9.7, and −8.7 kcal/mol, respectively, indicating their potential to modulate the antioxidant defense system through competitive inhibition of Keap1-Nrf2 interactions. This study provides a novel approach for the high-value utilization of bovine lung and the treatment of ALD, as well as a new source for the extraction of natural antioxidant peptides. Full article

(This article belongs to the Section Extraction and Industrial Applications of Antioxidants)

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21 pages, 1456 KB

Open AccessArticle

Comparative Pharmacokinetics and Safety of a Micellar Chrysin–Quercetin–Rutin Formulation: A Randomized Crossover Trial

by Afoke Ibi, Chuck Chang, Yun Chai Kuo, Yiming Zhang, Peony Do, Min Du, Yoon Seok Roh, Roland Gahler, Mary Hardy and Julia Solnier

Antioxidants 2025, 14(11), 1313; https://doi.org/10.3390/antiox14111313 - 31 Oct 2025

Abstract

Chrysin is a dietary flavonoid with antioxidant and anti-inflammatory activity, but its clinical potential is limited by poor oral bioavailability. This randomized double-blind three period crossover trial evaluated the pharmacokinetics of a novel micellar chrysin formulation co-encapsulated with quercetin and rutin (LMC) compared [...] Read more.

Chrysin is a dietary flavonoid with antioxidant and anti-inflammatory activity, but its clinical potential is limited by poor oral bioavailability. This randomized double-blind three period crossover trial evaluated the pharmacokinetics of a novel micellar chrysin formulation co-encapsulated with quercetin and rutin (LMC) compared with a non-micellar chrysin formulation (NMC) and unformulated chrysin (UFC). Secondary objectives included in vitro permeability (Caco-2) and a 30-day safety assessment of daily LMC supplementation. Sixteen healthy adults received a single oral dose of each formulation in randomized order separated by a 7-day washout. Plasma chrysin was quantified over 24 h to determine pharmacokinetic parameters. In vitro Caco-2 assays evaluated permeability, and clinical biochemistry of 15 participants were assessed weekly during 30 days of daily LMC use. LMC achieved >2-fold higher systemic exposure than unformulated chrysin (AUC_0–24 = 914.8 ± 697.5 ng·h/mL; C_max = 87.3 ± 59.4 ng/mL; both p < 0.05) and >2.6-fold higher than NMC, supported by >10-fold higher in vitro permeability. Daily LMC supplementation was well tolerated, with only mild, reversible adverse events and no clinically relevant safety changes, despite higher systemic exposure. Small, but significant, reductions in fasting glucose were observed in both sexes. The novel micellar chrysin–quercetin–rutin formulation substantially improved bioavailability and was well tolerated during 30 days of daily use, supporting its potential as an advanced delivery strategy for flavonoids with poor oral absorption and identifying glucose regulation as a physiological effect of interest. Full article

(This article belongs to the Special Issue Plant Polyphenols in Human Health: Emerging Insights for Future Therapeutic Strategies)

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19 pages, 8280 KB

Open AccessArticle

Pyrroloquinoline Quinone Mitigates Testicular Injury and Reduces Oxidative Stress, Mitochondrial Dysfunction, and Apoptosis in Rats with Testicular Ischemia–Reperfusion Injury

by Syuan-Hao Syu, Chao-Yuan Chang, Hung-Jen Shih and Chun-Jen Huang

Antioxidants 2025, 14(11), 1312; https://doi.org/10.3390/antiox14111312 - 31 Oct 2025

Abstract

Testicular torsion–detorsion (T/D) induces ischemia–reperfusion injury, leading to mitochondrial dysfunction, oxidative stress, apoptosis, and spermatogenic impairment. Pyrroloquinoline quinone (PQQ), a redox cofactor with mitochondrial-protective, antioxidant, and anti-apoptotic properties, was evaluated for its therapeutic potential in a rat T/D model. Young adult male Sprague-Dawley [...] Read more.

Testicular torsion–detorsion (T/D) induces ischemia–reperfusion injury, leading to mitochondrial dysfunction, oxidative stress, apoptosis, and spermatogenic impairment. Pyrroloquinoline quinone (PQQ), a redox cofactor with mitochondrial-protective, antioxidant, and anti-apoptotic properties, was evaluated for its therapeutic potential in a rat T/D model. Young adult male Sprague-Dawley rats underwent 720° spermatic cord rotation for 2 h followed by detorsion and were assigned to T/D or T/D + PQQ groups, with sham-operated controls run in parallel. PQQ (400 mg/kg body weight) was administered orally once daily for 4 weeks. T/D resulted in severe disruption of testicular architecture, disorganization of seminiferous epithelium, reduced sperm count and testis-to-body weight ratio, increased hypoxia-inducible factor-1α and malondialdehyde, decreased superoxide dismutase 2, impaired oxidative phosphorylation (OXPHOS), and enhanced apoptosis. Notably, PQQ treatment significantly preserved testicular structure, improved sperm counts, reduced oxidative stress, restored OXPHOS, and suppressed apoptosis (all p < 0.05. T/D + PQQ vs. T/D). These findings indicate that PQQ protects against T/D-induced testicular injury. The underlying mechanisms may involve the attenuation of oxidative stress, the preservation of mitochondrial function, and the limitation of apoptosis, supporting its potential as a therapeutic strategy for testicular IRI. Full article

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

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22 pages, 2404 KB

Open AccessArticle

Oxidative Stress and NLRP3 Inflammasome as Markers of Cardiac Injury Following Cardiopulmonary Bypass: Potential Implications for Patients with Preoperative Heart Failure with Reduced Ejection Fraction

by Rodrigo L. Castillo, Rodrigo A. Carrasco, Alejandro Gonzaléz-Candia, Esteban G. Figueroa, Adolfo A. Paz, Alejandro A. Candia, Sawa Kostin, Nikolaos Pagonas, Pamela V. Arias, Emilio A. Herrera, Robert A. Pérez and Sebastián Iturra

Antioxidants 2025, 14(11), 1311; https://doi.org/10.3390/antiox14111311 - 30 Oct 2025

Abstract

Cardiopulmonary bypass (CPB) can lead to cardiac damage due to oxidative stress (OS) and inflammation in heart failure (HF). We tested the hypothesis that preoperative HF patients with reduced ejection fraction (HFrEF) subjected to CBP have higher levels of OS and NLRP3 (NOD-, [...] Read more.

Cardiopulmonary bypass (CPB) can lead to cardiac damage due to oxidative stress (OS) and inflammation in heart failure (HF). We tested the hypothesis that preoperative HF patients with reduced ejection fraction (HFrEF) subjected to CBP have higher levels of OS and NLRP3 (NOD-, LRR- and pyrin domain-containing protein 3) in heart and plasma and in those that develop postoperative AF (pAF) as a clinical outcome. HF was categorized for preoperative left ventricular EF: preserved (HFpEF > 50%, n = 27) and reduced EF (HFrEF ≤ 40%, n = 25). Samples of atrial tissue, pericardial fluid, and plasma were collected at surgery to assess NLRP3 expression; 3-nitrotyrosine (3-NT), thiobarbituric acid reaction (TBARS), and nuclear factor erythroid 2-related factor 2 (Nrf2) in atrial tissue; NLRP3, IL-1β, and IL-18 expression in pericardial fluid; and antioxidant capacity, 8-isoprostanes, and malondialdehyde (MDA) in plasma. Reactive oxygen species, 3-NT, and NLRP3 in atrial tissue were determined by immunohistochemistry in a subset of pAF patients. Plasma and atrial tissue 3-NT and MDA were higher in HFrEF compared with HFpEF. Lipid peroxidation products were higher in both plasma and atrial tissue in pAF (n = 29), compared to sinus rhythm (SR) (n = 23). In HFrEF patients, the values of tissue ROS, 3-NT, and NLRP3 were higher than in HFpEF patients. In addition, the expression levels of NLRP3, IL-1β, and IL-18 were higher in atrial tissue and pericardial fluid in HFrEF. Patients with preoperative HFrEF showed higher OS in plasma and the expression of NLRP3, ROS, and 3-NT in atrial tissue biopsies and pericardial fluid. This finding suggests a potential pharmacologic therapy for pAF and clinical complications due to CPB. Full article

(This article belongs to the Special Issue Oxidative Stress in Cardiovascular Diseases (CVDs))

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22 pages, 6046 KB

Open AccessArticle

Deciphering the Class III Peroxidase Gene Family and Verifying Their Expression in Modulating Seed Germination in Tomato

by Jingbo Sun, Feng Zhang, Zhichao Zhao, Mengxia Zhang and Chunjuan Dong

Antioxidants 2025, 14(11), 1310; https://doi.org/10.3390/antiox14111310 - 30 Oct 2025

Abstract

Seed germination is crucial for seedling establishment and is regulated by precise reactive oxygen species (ROS) signaling. Class III peroxidases (PRXs), which are plant-specific enzymes, play crucial roles in plant growth, development, and responses to abiotic stress by maintaining ROS homeostasis. However, members [...] Read more.

Seed germination is crucial for seedling establishment and is regulated by precise reactive oxygen species (ROS) signaling. Class III peroxidases (PRXs), which are plant-specific enzymes, play crucial roles in plant growth, development, and responses to abiotic stress by maintaining ROS homeostasis. However, members of the PRX gene family in tomato, particularly their functions in modulating seed germination, remain poorly understood. In this study, 102 tomato PRXs (SlPRXs) were identified, and they were classified into five groups based on phylogenic analysis. Chromosomal localization revealed that these SlPRX genes are unevenly distributed across 12 tomato chromosomes, with chromosome 02 harboring the highest densities. Gene structure analysis revealed that SlPRXs contain 1 to 10 exons, and SlPRX4 possesses the most exons. All SlPRX proteins possess the characteristic peroxidase domain and share conserved structural motifs. Collinearity analysis suggested that segmental duplications might be the main contributor to the expansion of the SlPRX family. Promoter analysis revealed numerous cis-acting elements related to abiotic/biotic stress responses, phytohormones, and growth and development. Notably, seed germination-related elements such as CARE and RY element were identified in some SlPRXs. Enzymatic and electrophoresis assays indicated that PRX activity increased with seed germination. Moreover, SHAM, the inhibitor of PRX, exerted an inhibitory effect on tomato seed germination. Transcriptome data revealed stage-specific induction of SlPRXs during germination, with distinct expression peaks between 0 and 96 h post imbibition. These findings were further validated by qRT-PCR of the selected SlPRX genes. Overall, the findings enhance our understanding of SlPRX family members in tomato and highlight their potential for improving seed germination. This study also provides valuable genetic resources and potential molecular markers for breeding tomato varieties with improved germination vigor and stress resilience. Full article

(This article belongs to the Section Natural and Synthetic Antioxidants)

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27 pages, 6344 KB

Open AccessArticle

Effects of Black Soldier Fly (Hermetia illucens) Larvae Meal Replacement for Fish Meal on Growth Performance, Muscle Quality, Antioxidant Status, Immune Function, and Gut Microbiota in Juvenile Southern Catfish (Silurus meridionalis)

by Huiying Wang, Gao Gao, Jialong Chen, Dan Jia, Qing Hu, Hanqi Duan, Bin Zhang, Run Bi, Qingquan Hu and Baoliang Bi

Antioxidants 2025, 14(11), 1309; https://doi.org/10.3390/antiox14111309 - 30 Oct 2025

Abstract

This study evaluated the effects of feeding juvenile southern catfish (Silurus meridionalis) with one of six isonitrogenous and isoenergetic diets where fish meal (FM) was replaced by black soldier fly larval meal (BSFLM) at 0%, 10%, 20%, 30%, 40%, and 50% [...] Read more.

This study evaluated the effects of feeding juvenile southern catfish (Silurus meridionalis) with one of six isonitrogenous and isoenergetic diets where fish meal (FM) was replaced by black soldier fly larval meal (BSFLM) at 0%, 10%, 20%, 30%, 40%, and 50% levels on growth, muscle quality, antioxidant capacity, immune response, and gut microbiota of juvenile southern catfish (Silurus meridionalis). A total of 1620 fish (9.20 ± 0.15 g) were fed one of six experimental diets for 8 weeks. Results demonstrated that a 50% replacement (H50 group) significantly improved weight gain rate, specific growth rate, and protein efficiency ratio (p < 0.001). Antioxidant capacity (T-AOC) was enhanced in groups H30 and H50, while immune markers lysozyme (LZM) and alkaline phosphatase (AKP) showed mixed responses. Muscle texture properties such as chewiness and adhesiveness were significantly altered across treatments. Gut villi remained structurally intact in all groups, and liver histology appeared normal. No significant differences were found in muscle amino acid or fatty acid profiles. Gut microbiota analysis revealed shifts in microbial composition, with increased abundance of Clostridia and Escherichia and functional enrichment in metabolic pathways at higher substitution levels. Interspecies network analysis indicated potential cooperation among beneficial microbes through metabolite exchange. It is concluded that 50% BSFLM substitution optimizes growth performance, muscle quality, and antioxidant capacity, while modulating gut microbiota, indicating its promise as a sustainable FM alternative and functional ingredient in aquafeeds. Full article

(This article belongs to the Special Issue Antioxidant Properties in Novel Feed Ingredients for Fish)

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19 pages, 1189 KB

Open AccessArticle

Genus Veronica—Antioxidant, Cytotoxic and Antibacterial Activity of Phenolic Compounds from Wild and Cultivated Species

by Ivana Vrca, Antonija Mikrut, Željana Fredotović, Karla Akrap, Dario Kremer, Stjepan Orhanović, Katarina Bačić, Valerija Dunkić and Marija Nazlić

Antioxidants 2025, 14(11), 1308; https://doi.org/10.3390/antiox14111308 - 30 Oct 2025

Abstract

(1) Background: The conservation of plant resources is important because many wild plant populations are threatened by various influences. Growing plants from seeds is one way to ensure their survival. Comparing the biological potential of extracts between plants cultivated from seeds and wild [...] Read more.

(1) Background: The conservation of plant resources is important because many wild plant populations are threatened by various influences. Growing plants from seeds is one way to ensure their survival. Comparing the biological potential of extracts between plants cultivated from seeds and wild plants provides information about their specialized metabolites. For this reason, this study compared the biological potential of phenolic extracts from four wild-collected species of the genus Veronica and the same four cultivated species. The studied species of the genus Veronica are V. anagallis-aquatica L., V. persica Poir., V. polita Fr. and V. hederifolia L. (2) Methods: The phenolic composition was investigated with LC-QTOF (Liquid Chromatography-Quadrupole Time-of-Flight). The main methods for biological activities were as follows: ORAC (Oxygen Radical Absorbance Capacity) and DPPH (2,2-diphenyl-1-picrylhydrazyl) radical for antioxidant activity, the disk diffusion test for antibacterial activity and the MTS test of cytotoxic activity. (3) Results: The major compound in all extracts was apigenin. Cultivated species showed higher antioxidative activity. Phenolic compounds isolated from the V. anagallis-aquatica species showed the highest cytotoxic effect on all tested lines. The extracts showed antibacterial activity on three bacterial strains: Streptococcus pyogenes, Listeria monocytogenes and L. innocua. Extracts of V. anagallis-aquatica showed the highest antibacterial activity, both from the natural habitat and cultivated habitat. (4) Conclusions: A comparison of the different activities tested for phenolic extracts between wild and cultivated species of the genus Veronica showed that cultivated species also have significant biological activity and are suitable for further research and applications. Full article

(This article belongs to the Special Issue Phytochemical Profiling, Antioxidant Activity and Therapeutic Properties of Medicinal Plants)

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21 pages, 2976 KB

Open AccessArticle

In Vivo Studies on the Interaction Between Orally Administered Nitrite and Omeprazole: Beyond Proton-Catalyzed S-Nitrosation

by Macario A. Rebelo, Alessandra Cássia-Barros, Sandra O. Conde-Tella, Sabrina F. Frugeri, Paula P. Ovidio, Alceu A. Jordão Junior, Cezar Kayzuka, Riccardo Lacchini, Alessandra O. Silva, Carlos R. Tirapellli, Martin Feelisch and Jose E. Tanus-Santos

Antioxidants 2025, 14(11), 1307; https://doi.org/10.3390/antiox14111307 - 30 Oct 2025

Abstract

Inorganic nitrite contributes to the nitrosation of biomolecules and exerts antioxidant effects. The proton pump inhibitor omeprazole has pro-oxidant effects, inhibits the formation of nitroso species in the stomach, and abrogates the blood pressure-lowering effects of orally administered nitrite. Here, we examine whether [...] Read more.

Inorganic nitrite contributes to the nitrosation of biomolecules and exerts antioxidant effects. The proton pump inhibitor omeprazole has pro-oxidant effects, inhibits the formation of nitroso species in the stomach, and abrogates the blood pressure-lowering effects of orally administered nitrite. Here, we examine whether a two-week treatment with nitrite leads to tissue nitrosation that scales with local thiol concentrations and whether oral nitrite treatment can prevent the pro-oxidant effects of omeprazole. Male Sprague–Dawley rats received daily doses of omeprazole 10 mg/kg i.p. (or vehicle) and sodium nitrite 15 mg/kg by gavage (or water) for 14 days. Animals were euthanized 6 h after the last nitrite dose, and blood and tissues (brain, heart, and liver) were collected for biochemical analyses. We found that nitrite treatment increased liver nitrite and total nitroso species (RxNO) concentrations approximately eight-fold (with minor increases in other organs), and omeprazole treatment attenuated these effects. Nitrite treatment selectively elevated non-protein thiol concentrations in the liver, but not in animals also receiving omeprazole. Tissue thiol elevation was associated with increased nitrosation of hepatic proteins, which was prevented by omeprazole. Nitrite upregulated mRNA expression of microsomal glutathione S-transferase-1 (Mgst1) and decreased superoxide and hydrogen peroxide production, especially in rats co-treated with omeprazole. While omeprazole increased liver xanthine oxidoreductase (XOR), nitrite treatment attenuated this effect. These results demonstrate that oral nitrite treatment robustly elevates nitrite and RxNO concentrations in the liver, and these effects are associated with increased hepatic glutathione production and an upregulation of Mgst1 expression, counteracting the pro-oxidant effects induced by omeprazole. Full article

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

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17 pages, 2299 KB

Open AccessArticle

Reactive Oxygen Species (ROS) Drive Osteocyte Dysfunction in Diabetic Osteoporosis by Impairing Autophagy and Triggering Apoptosis

by Mengqi Han, Minyue Zhao, Furong Bai, Mengying Wang, Bo Zhang, Jianfeng Shi and Zhongbo Liu

Antioxidants 2025, 14(11), 1306; https://doi.org/10.3390/antiox14111306 - 30 Oct 2025

Abstract

This study investigates the mechanisms underlying osteocyte injury in a high glucose (HG) environment and explores potential therapeutic targets and diagnostic markers for diabetic osteoporosis, a common complication of type 2 diabetes mellitus (T2DM). Hyperglycemia induces oxidative stress through the reactive oxygen species [...] Read more.

This study investigates the mechanisms underlying osteocyte injury in a high glucose (HG) environment and explores potential therapeutic targets and diagnostic markers for diabetic osteoporosis, a common complication of type 2 diabetes mellitus (T2DM). Hyperglycemia induces oxidative stress through the reactive oxygen species (ROS) production, which impair osteocytes and accelerate bone loss. To examine these effects, MLO-Y4 cells and primary mouse osteocytes were cultured under normal glucose and HG conditions, with additional treatments using N-acetylcysteine (NAC, ROS scavenger) and rapamycin (autophagy promoter and mTOR inhibitor). Cell viability, ROS levels, and the autophagy and apoptosis markers expression (Beclin1, LC3, p62, Bax, Bcl2, cytochrome C, and caspase3) were assessed using CCK8/ATP level assay, flow cytometry, Western blot, qRT-PCR, immunofluorescence, and TUNEL staining. The results showed that HG inhibits cell proliferation, induces insulin resistance, generates ROS, alters antioxidant enzymes, and promotes oxidative stress, leading to mTOR activation, subsequent autophagy inhibition, and osteocyte apoptosis. NAC mitigated these effects, while rapamycin prevented HG-induced apoptosis by inhibiting mTOR activation and promoting autophagy. This suggests that ROS-induced mTOR activation impairs autophagy and hinders the clearance of damaged osteocytes, triggering apoptosis. This research provides foundational evidence and novel insights into diabetic osteoporosis pathogenesis and potential therapies. Full article

(This article belongs to the Special Issue In Honor of Jiankang Liu for His Contributions and Impact on Redox and Mitochondrial Biology and Medicine)

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14 pages, 1101 KB

Open AccessArticle

Determinants of Outcome Variability in Ischemic Stroke: A Focus on Routinely Collected Biomarkers

by Alexandru Gerdanovics, Sorana D. Bolboacă, Ioana Cristina Stănescu, Camelia Manuela Mîrza, Gabriela Bombonica Dogaru, Cristina Ariadna Nicula, Paul Mihai Boarescu, Cezara-Andreea Gerdanovics and Adriana-Elena Bulboacă

Antioxidants 2025, 14(11), 1305; https://doi.org/10.3390/antiox14111305 - 30 Oct 2025

Abstract

Ischemic stroke remains a leading cause of mortality and disability, with proinflammatory, metabolic, and oxidative stress-related factors contributing to outcome variability. We conducted a retrospective cross-sectional study of 124 consecutive patients (53 women, 71 men; median age 71 [62–76]) discharged with ICD-10 code [...] Read more.

Ischemic stroke remains a leading cause of mortality and disability, with proinflammatory, metabolic, and oxidative stress-related factors contributing to outcome variability. We conducted a retrospective cross-sectional study of 124 consecutive patients (53 women, 71 men; median age 71 [62–76]) discharged with ICD-10 code I69.3 from the Neurology Department of the Clinical Rehabilitation Hospital in Cluj-Napoca (January 2023–September 2024). Men were younger (median age of 69 vs. 73 years, p-value = 0.010), more frequently smokers (42% vs. 9%, p < 0.001), and alcohol consumers (21% vs. 4%, p-value = 0.007) than women. In contrast, women were more frequently sedentary (68% vs. 49%, p-value = 0.038) and had higher LDL cholesterol (89 vs. 74 mg/dL, p = 0.026) than men. Patients with at least moderate disability (n = 84) presented higher levels of C-Reactive Protein (CRP), 1.4 vs. 1.1 mg/L, p-value = 0.027) and more frequently low HDL cholesterol serum levels (29.8% vs. 7.5%, p-value = 0.006) compared to those with minor disability. In multivariable regression, low HDL was the sole independent predictor of disability severity (OR = 4.58, 95% CI 1.21–17.41; AUC = 0.78, sensitivity = 88%, specificity = 42%), while CRP and age did not retain the significance obtained in univariable regression. Our findings highlight sex-specific risk profiles and underline the contribution of proinflammatory, metabolic, and oxidative pathways to ischemic stroke severity, underscoring the need for prospective validation in larger cohorts. Full article

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

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