Antioxidants

Peripheral blood measurements dominate oxidative stress research, yet whether they reflect central nervous system (CNS) redox status remains untested in humans. We simultaneously profiled five biomarkers, total antioxidant capacity (TAC), glutathione (GSH), thiobarbituric acid-reactive substances (TBARS), ferric reducing antioxidant power (FRAP), and hydroxyl radical scavenging activity (HRSA), in paired blood and cerebrospinal fluid (CSF) from 140 adults in the ALBION cohort. Only FRAP showed a significant positive cross-compartment correlation ({rho} = +0.49, FDR-p < 0.001), supporting its role as a systemic antioxidant signal. TBARS showed a significant inverse cross-compartment association ({rho} = -0.20, FDR-p = 0.042), suggesting compartmental compensation in lipid peroxidation regulation rather than parallel dynamics. TAC and GSH showed no meaningful intercompartmental alignment. Individual biomarker levels were largely stable across the 40-85 year age range in both compartments, suggesting that age effects operate through coordinated latent networks rather than single-marker trajectories. Principal component extraction with varimax rotation identified four latent factors explaining 66.6% of total variance, dominated by a coherent CSF-centred redox axis alongside multiple partially opposing peripheral components. Age stratification revealed progressive fragmentation: middle-aged adults retained four coherent cross-compartment factors, whereas older adults exhibited five more dispersed components. Sex-stratified analyses showed that females exhibited four-factor modular organisation centred on glutathione, while males showed a simpler three-factor structure with tighter cross-compartment coupling anchored by FRAP. Blood and CSF oxidative stress biomarkers are not interchangeable, a finding with direct implications for biomarker selection in clinical trials targeting neurological conditions.

The evidence linking high levels of environmental pollutants to chronic degenerative diseases is alarming, with heavy metals (HM) identified as a key factor. Research suggests that certain phytochemicals in the diet can reduce HM levels and mitigate their adverse health effects.Curcumin (Cur), a natural polyphenol, is particularly effective in protecting against Cadmium (Cd) toxicity. The present study demonstrates that preincubation with low doses of Cur (1 M) in differentiated HL-60 and K-562 human myeloid cells can significantly protect against cytotoxicity induced by Cd and or H2O2. Cur reduced the increased levels of reactive oxygen species (ROS) generated by Cd or H2O2 by inducing a protective form of autophagy. Cur activated mild oxidative stress that triggers the expression of Nrf2-dependent transcripts, such as HO and NQO1. The potential chemopreventive effects of Cur against redox stress have been strengthened by the observation that free and unmetabolized Cur is detectable inside the cells after 5 minutes of treatment, and its presence parallels with increased levels of intracellular GSH. These findings suggest that supplementation with Cur in the form of nutraceuticals may represent a promising way to protect people living in highly polluted areas against the adverse effects of HM contaminants.

Xeroderma pigmentosum group C fiborblasts (XP-C) are characterized by chronic redox imbalance and elevated H{square}O{square}levels, making them a good model for testing compounds with antioxidant potential for therapeutic purposes. Here, we investigated the effects of ebselen, a compound with glutathione peroxidase (GPx) mimetic activity, in the XP-C model. We found that ebselen behaves as a hormetic compound, protecting cells against H2O2-induced cytotoxicity at low doses but potentiating the cytotoxic effect at higher doses. Accordingly, when administered chronically, ebselen significantly reduces H{square}O{square}production and p53 levels. However, acute treatment with ebselen causes a reduction in O2 consumption (OCR) and extracellular acidification rate (ECAR), indicative of decreased mitochondrial function and metabolic activity. In addition, acute ebselen treatment causes a reduction in the GSH/GSSG ratio and an increase in NRF-2 expression, suggesting that ebselen induces redox stress that triggers an adaptive response, characterizing a possible mitohormetic effect. The reduction in the GSH/GSSG ratio appears to be the initial trigger after acute treatment with ebselen, since concomitant treatment with NAC prevents the reduction in OCR, ECAR and NRF-2 activation, in addition to protecting XP-C cells against lethal doses of ebselen. HighlightsO_LIChronic ebselen treatment reduces H2O2 levels, lowers p53 expression and protects XP-C cells against oxidative insults. C_LIO_LIInitially, ebselen causes mitochondrial stress, which is followed by cellular adaptation and protection against oxidative stress. C_LIO_LIEbselen acts as a hormetic drug, likely through a mitohormetic mechanism. C_LI

Melatonin has emerged as a promising pharmacological candidate for bipolar disorder (BD), though its mechanisms of action remain incompletely understood. Its antioxidant, anti-inflammatory, and anti-dopaminergic properties suggest potential relevance to BD pathophysiology. This study investigated melatonins effects on dopamine signalling, metabolism, and oxidative stress under inflammatory and hyperdopaminergic conditions in differentiated SH-SY5Y neuronal cells. Cells were pretreated with 100nM melatonin or vehicle for 2 hours, then exposed to vehicle, IL-6 (20ng/mL), dopamine (5{micro}M or 500{micro}M), or dopamine (500{micro}M) with ascorbic acid (1mM) for 12 or 24 hours. Dopaminergic markers were assessed via real-time PCR and HPLC; metabolic outcomes were measured using Seahorse assay, central carbon metabolomics, in-cell Western assay, and glucose uptake assay; and oxidative stress was evaluated via reactive oxygen species (ROS), superoxide (SOX), and total antioxidant capacity (TAC) assays. IL-6 increased dopamine levels, p-Erk1/2/Erk1/2, p-AMPK/AMPK, nucleotide pools, and TAC, while reducing dopamine turnover, SV2C expression, and spare respiratory capacity. Melatonin alone increased nucleotides and NADH, while reducing dopamine turnover, ROS, and glucose-1-phosphate. In IL-6 conditions, melatonin pretreatment enhanced spare respiratory capacity, glucose uptake, and NADH, while reducing dopamine, TAC, p-AMPK/AMPK, p-GSK3{beta}/GSK3{beta}, and non-mitochondrial oxygen consumption. High-dose dopamine (500{micro}M) elevated SOX, p-Erk1/2/Erk1/2, insulin receptor-, GLUT1, glycolytic ATP (glycoATP), and non-mitochondrial oxygen consumption. Melatonin pretreatment attenuated p-Erk1/2/Erk1/2 and GLUT1 elevations. Combined dopamine and ascorbic acid further increased glycolytic intermediates, ROS, p-AMPK/AMPK, and TAC, while reducing p-Erk1/2/Erk1/2, p-mTOR, GLUT1, glucose uptake, and glycoATP. Overall, melatonin mitigated IL-6-induced dopaminergic, oxidative, and metabolic alterations, and partially protected against dopamine-induced metabolic shifts. These findings suggest melatonin may alleviate manic symptoms in BD via both direct dopaminergic modulation and indirect antioxidant and metabolic regulatory effects.

The present study investigated the impact of mercury exposure on plant water status, oxidative stress markers, and antioxidant defense systems in leaves and fruits of tomato (Solanum lycopersicum L. cv. Micro-Tom). Plants were exposed to increasing concentrations of HgCl2 for 24 and 48 h. Mercury treatment led to a significant reduction in predawn leaf water potential, whereas other water-related parameters in both leaves and fruits remained largely unaffected. Oxidative stress was predominantly observed in leaves, as indicated by elevated hydrogen peroxide (H2O2) and malondialdehyde (MDA) levels, while fruit tissues showed no significant accumulation of these oxidative markers. In contrast, fruits exhibited a marked activation of antioxidant defenses, including increased activities of superoxide dismutase (SOD) and catalase (CAT), along with concentration-dependent modulation of ascorbate-glutathione cycle enzymes (APX, MDHAR, DHAR, and GR) and their corresponding transcript levels. Alterations in the ascorbate pool, reflected by changes in reduced ascorbate (AsA) and dehydroascorbate (DHA), further indicated a dynamic regulation of cellular redox status in response to mercury exposure. Collectively, these findings demonstrate that mercury induces tissue-specific oxidative responses and rapidly activates antioxidant mechanisms in fruits, thereby contributing to the maintenance of redox homeostasis and protection against oxidative damage.

Markers of oxidative stress are widely used as biomarkers of health, ageing, and physiological stress. However, their reliability as biomarkers remains uncertain due to high intraindividual variation, obscuring associations with environmental conditions, lifestyle, frailty, and physiological indicators of health such as telomere length. Whilst numerous longitudinal studies exist, individual repeatability of oxidative stress measurements is rarely reported. This study presents the first meta-analysis assessing individual repeatability of oxidative stress markers, comprising 123 repeatability estimates from 22 studies. We found that oxidative stress exhibits low individual repeatability (Intraclass correlation = 0.164), regardless of marker type, taxa, sex, study design, or environment. We also found that different markers of oxidative stress were often poorly correlated, which could be due to their low repeatability. This flags serious limitations regarding the utility of oxidative stress measurements as health biomarkers. To illustrate this point, we simulated causal effects of oxidative stress on telomere length to reveal statistical power limitations on the detection of a relationship between oxidative stress and telomere length when individual repeatability is low. Our simulations reveal that substantially larger samples sizes are required than those typically used in this field. On a more positive note, we also show that increasing the number of repeated samples can improve statistical power. Such longitudinal studies would create valuable opportunities for untangling the causes of intraindividual variation in oxidative stress, elucidating the functional consequences of oxidative stress for health and ageing.

Glutathione is the most abundant thiol in animal cells. Reduced glutathione (GSH) is a major intracellular antioxidant neutralizing free radicals and detoxifying electrophiles. It plays important roles in many cellular processes, including cell differentiation, proliferation, and apoptosis. In the present study we demonstrate that extracellular concentration of reduced glutathione markedly increases cell volume within few hours, in a dose-response manner. Pre-incubation of cells with BSO, the inhibitor of {gamma}-glutamylcysteine synthetase, responsible for the first step in intracellular glutathione synthesis did not change the effect of reduced glutathione on cell volume suggesting a mechanism limited to the interaction of extracellular reduced glutathione on cell membrane. Similarly, inhibition of {gamma}-glutamylcyclotransferase involved in intracellular glutamate production had no effect on the action of reduced glutathione. Oxidized glutathione exerted no effect on cell volume. Results show that reduced GSH decreases cell adhesion resulting in an increased cell volume. Since many cell types are able to export GSH, the present results suggest that this could be a fundamental self-regulation of cell volume, giving the cells a self-control on their adhesion proteins.

Background and ObjectivesExcessive Manganese (Mn) exposure is neurotoxic and can cause Mn-Induced Parkinsonism (MnIP), marked by cognitive and motor dysfunction. Although metabolomic and lipidomic research in Parkinsonism (PD) patients exists, it remains limited. This study hypothesizes distinct metabolomic and lipidomic profiles based on exposure status, disease diagnosis, and their interaction. MethodsWe used a case-control design with a 2x2 factorial framework to investigate the metabolomic and lipidomic alterations associated with Mn exposure and their link to PD. The study population of 97 individuals was divided into four groups: non-exposed controls (n=23), exposed controls (n=25), non-exposed with PD (n=26) and exposed with PD (n=23). Cases, defined by at least two cardinal PD features (excluding vascular, iatrogenic, and traumatic origins), were recruited from movement disorder clinics in four hospitals in Brescia, Northern Italy. Controls, free from neurological or psychiatric conditions, were selected from the same hospitals. Exposed subjects resided in metallurgic regions (Val Camonica and Bagnolo Mella) for at least 8 continuous years, while non-exposed subjects lived in low-exposure areas around Lake Garda and Brescia city. We conducted untargeted analyses of metabolites and lipids in whole blood samples using ultra-high-performance liquid chromatography (UHPLC) and mass spectrometry (MS), followed by statistical analyses including Principal Component Analysis (PCA), Partial Least Squares-Discriminant Analysis (PLS-DA), and Two-Way Analysis of Covariance (ANCOVA). ResultsMetabolomic analysis revealed modulation of alanine, aspartate, and glutamate metabolism (Impact=0.05, p=0.001) associated with disease effect; butanoate metabolism (Impact=0.03, p=0.004) with the exposure effect; and vitamin B6 metabolism (Impact=0.08, p=0.03) with the interaction effect. Differential relative abundances in 3- sulfoxy-L-Tyrosine ({beta}=1.12, FDR p<0.001), glycocholic acid ({beta}=0.48, FDR p=0.03), and palmitelaidic acid ({beta}=0.30, FDR p<0.001) were linked to disease, exposure, and interaction effects, respectively. In the lipidome, ferroptosis (Pathway Lipids=11, FDR p=0.03) associated with the disease effect and sphingolipid signaling (Pathway Lipids=9, FDR p=0.04) associated with the interaction effect were significantly altered. Lipid classes triacylglycerols, ceramides, and phosphatidylethanolamines showed differential relative abundances associated with disease, exposure, and interaction effects, respectively. DiscussionThese findings suggest that PD and Mn exposure induce unique metabolomic and lipidomic changes, potentially serving as biomarkers for MnIP and warranting further study.

Geographic atrophy or late stage dry age-related macular degeneration (AMD) is characterized by drusen deposition and progressive retinal pigment epithelium (RPE) degeneration, leading to irreversible vision loss. The formation of drusen leads to dyshomeostasis, oxidative stress and irreversible damage to RPE. In this study, we used an in vitro model of oxidized-low density lipoproteins (ox-LDL) induced human RPE damage/death model to investigate the mechanism whereby a sterically hindered phenol antioxidant compound, PMC (2,2,5,7,8-pentamethyl-6-chromanol) protects RPE against ox-LDL-induced damage. We show that PMC exerts its protective effect by preventing the upregulation of stress-responsive heme oxygenase-1 (HMOX1/HO-1) and NAD(P)H:quinone oxidoreductase (NQO1) at mRNA and protein levels. This effect was due to PMCs blockade of ROS generation, which in turn blocked nuclear translocation of the Nuclear factor erythroid 2-related factor 2 (Nrf2) transcription factor, ultimately preventing the upregulation of antioxidant response elements (ARE), including HMOX1 and NQO1. A key role for HO-1 was demonstrated when the protective effect of PMC was inhibited by the knockdown of HMOX1. Additionally, treatment of PMC under different experimental conditions and time points revealed that the continuous presence of PMC is required for optimal protection against ox-LDL-induced cytotoxicity, defining the cellular pharmacokinetics of the molecule. Our data demonstrate the involvement of a key antioxidant pathway through which PMC mitigates oxidative stress induced by ox-LDL and provides a potential therapeutic strategy to suppress RPE degeneration/damage during AMD progression.

Reactive oxygen species (ROS) play a dual role in cellular homeostasis, but excessive levels of ROS lead to oxidative stress, accelerating skin aging. Environmental stressors like UV radiation induce ROS overproduction, overwhelming endogenous antioxidant defenses and causing cellular damage. While the skin possesses an intrinsic antioxidant network that provides moderate protection, excessive oxidative stress can trigger inflammatory responses, thereby necessitating exogenous antioxidant intervention. Microbe-derived antioxidants (MA), produced via probiotic fermentation of sea buckthorn and chestnut rose, have shown promise in mitigating ROS-induced damage. In this study, we evaluated two MA formulations, MA1 and MA2, for their ability to scavenge free radicals and alleviate hydrogen peroxide (H2O2)-induced oxidative stress in human dermal fibroblasts (HDF) and dermal papilla cells (HDP). Both formulations displayed dose-dependent DPPH radical scavenging activity and enhanced cell viability at low concentrations. Under H2O2-induced oxidative stress, MA1 and MA2 effectively restored intracellular ROS to baseline levels, demonstrating significant cytoprotective effects. UHPLC-MS/MS profiling identified 12 compounds shared by both formulations, and Gene Ontology Biological Process enrichment analysis revealed that their associated target genes were significantly enriched in antioxidant-related pathways. Five compounds--adenosine, citric acid, 5-hydroxymethylfurfural, myricetin, and phenylalanine--emerged as key contributors to the observed antioxidative effects. Together, these findings highlight the potential of fermented microbial antioxidants to re-establish redox homeostasis in human skin cells and support their further development as therapeutic or cosmetic interventions targeting oxidative stress and skin aging. Given the heightened oxidative sensitivity of aged fibroblasts, MAs ability to alleviate ROS may offer novel therapeutic strategies against skin aging and related pathologies.

Since oxidative stress is a critical common pathological factor of numerous diseases, it is critical to find biomarkers for non-invasive evaluations of the levels of oxidative stress in the body. Our previous studies have indicated that epidermal green autofluorescence (AF) is a novel biomarker of this type: The oxidative stress inducer buthionine sulfoximine (BSO) can dose-dependently increase the epidermal green AF of mice, with BSO doses being significantly associated with the AF intensity. However, it is necessary to use skin cell cultures to investigate the mechanisms underlying the relationships between BSO and the green AF intensity. In our current study we found that BSO concentration-dependently increased the green AF intensity of B16-F10 cells a skin cell line, with BSO concentrations being significantly associated with the AF intensity. BSO also concentration-dependently increased the intracellular DCF signals an index of ROS levels. The green AF intensity of the cells was also significantly associated with the intracellular ROS levels. Moreover, we found that the green AF intensity was significantly associated with the cell death induced by BSO. Collectively, our study has provided first evidence indicating that the green AF intensity of skin cells is significantly associated with both intracellular ROS levels and cell death of the skin cells exposed to oxidative stress, which has indicated that green AF is a novel biomarker for both oxidative stress and cell death.

In previous studies, the oil extracted from the visceral fat of Caiman yacare (Daudin, 1802) demonstrated a wound-healing effect on the skin of Wistar rats. To enhance knowledge our about the mechanism underlying this effect, we analysed the oils toxicological potential in vitro. Cytotoxicity, genotoxicity, pro-oxidant, and antioxidant activities were evaluated in a V79-4 cell line. The oil was obtained using the Soxhlet method, and the proportions of the fatty acid profile was previously identified 43.74 % saturated and 34.65 % unsaturated fatty acids. Protocol 487 of the Organisation for Economic Co-operation and Development (OECD) was employed for cell line selection and concentrations. Cytotoxicity was determined using the MTT assay and clonogenic survival. Pro-oxidant and antioxidant activities were analysed using flow cytometry. Genotoxicity was evaluated using comet and micronucleus assays. The oil did not demonstrate cytotoxicity up to a concentration of 500 {micro}g/mL. At concentrations of 250 and 500 {micro}g/mL, the oil exerted a protective effect against oxidative stress and showed genotoxic effects only at the highest concentration (2000 {micro}g/mL). Like other oils of interest for human health, the oil extracted from the visceral fat of C. yacare demonstrated low toxicological potential in vitro. SUMMARY STATEMENTThe oil from Caiman yacare visceral fat presents low cytotoxicity and genotoxicity, highlighting its potential for therapeutic applications without adverse effects.

Iron superoxide dismutase 1 (FSD1) was recently characterized as a plastidial, cytoplasmic, and nuclear superoxide dismutase with osmoprotective and antioxidative functions. However, its role in oxidative stress tolerance is not well understood. Here, we characterized the role of FSD1 in response to methyl viologen (MV)-induced oxidative stress in Arabidopsis thaliana. The findings demonstrated that the antioxidative function of FSD1 depends on the availability of Cu2+ in growth media. Prolonged MV exposure led to a decreased accumulation rate of superoxide, higher levels of hydrogen peroxide production, and higher protein carbonylation in the fsd1 mutants and transgenic plants lacking a plastidial pool of FSD1, compared to the wild type. MV led to a rapid increase in FSD1 activity, followed by a decrease. Chloroplastic localization of FSD1 is necessary for these changes. Proteomic analysis showed that the sensitivity of the fsd1 mutants coincided with decreased abundance of ferredoxin and light PSII harvesting complex proteins, with altered levels of signaling proteins. Collectively, the study provides evidence for the conditional antioxidative function of FSD1 and its possible role in signaling.

Cellular energy metabolism varies depending on tissue and cell type, as well as the availability of energy substrates and energy demands. We recently investigated the variations in cellular metabolism and antioxidant responses in primary bovine vascular endothelial cells (BAECs) under different energetic substrate conditions in vitro, specifically glucose or galactose. In this context, pharmacological agents may affect cells differently depending on their energy metabolism status. In this study, we aimed to characterize the effects of diphenyl diselenide ((PhSe)2), a redox-active molecule known for its prominent cardiovascular effects, on redox-bioenergetic cellular pathways under glycolytic or oxidative conditions in BAECs. Under glucose conditions, (PhSe)2 positively impacted mitochondrial oxidative capacity, as assessed by respirometry, and was associated with changes in mitochondrial cellular dynamics. However, these changes were not observed in cells cultured with galactose. Although (PhSe)2 induced the nuclear translocation of the redox sensitive nuclear factor erythroid 2-related factor 2 (Nrf2) in both glucose and galactose media, Nrf2 remained in the nuclei of cells cultured in galactose for a longer duration. Additionally, activation of another redox sensitive transcription factor, forkhead O3 (FOXO3a) was only detected in galactose media. Notably, (PhSe)2 induced the expression of genes controlling mitochondrial antioxidant capacity and glutathione synthesis and recycling in glucose media, whereas its effects in galactose media were primarily focused on glutathione homeostasis. In conclusion, our findings underscore the critical influence of cellular metabolic status on the antioxidant capacity of redox-active molecules such as (PhSe)2.

Cells respond to oxidative stress by elevating the levels of antioxidants, signaling, and transcriptional regulation often implemented by chromatin remodeling proteins. The study presented in this paper shows that the expression of PICH, an ATP-dependent chromatin remodeler, is upregulated during oxidative stress in HeLa cells. We also show that PICH regulates the expression of Nrf2, a transcription factor regulating antioxidant response, both in the absence and presence of oxidative stress. In turn, Nrf2 regulates the expression of PICH in the presence of oxidative stress. Both PICH and Nrf2 together regulate the expression of antioxidant genes and this transcriptional regulation is dependent on the ATPase activity of PICH. In addition, H3K27ac modification also plays a role in activating transcription in the presence of oxidative stress. Co-immunoprecipitation experiments show that PICH and Nrf2 interact with H3K27ac in the presence of oxidative stress. Mechanistically, PICH recognizes ARE sequences present on its target genes and introduces a conformational change to the DNA sequences leading us to hypothesize that PICH regulates transcription by remodeling DNA. PICH ablation leads to reduced expression of Nrf2 and impaired antioxidant response leading to increased ROS content, thus, showing PICH is essential for the cell to respond to oxidative stress.

The role of foods is shifting from focusing on the provision of energy and basic nutrients to also include bioactive compounds that prevent the development of chronic age-related diseases. Microalgae offer a source of nutritional compounds with important health effects, and have the potential for production of health-promoting products, without increasing agricultural land use and negatively impacting the environment. Here we investigate the health effects of microalgal extracts with high levels of the colourless carotenoid phytoene. Phytoene is widely available from dietary sources and can be detected across different tissues in the human body. However, it is usually regarded as a precursor for the synthesis of other carotenoids, rather than a bioactive molecule. We utilised the model organism C. elegans to show that phytoene-rich extracts from Chlorella sorokiniana and Dunaliella bardawil have anti-ageing properties. The extracts protect against oxidative damage and amyloid-{beta}42 proteotoxicity (a major pathology of Alzheimers disease), and extends lifespan. We show that pure phytoene also has these anti-ageing effects, suggesting that phytoene is a bioactive molecule with positive effects on ageing and longevity.

Chronic inflammatory and metabolic diseases are major global health issues increasingly linked to dietary factors. Consumption of dietary emulsifiers like polysorbate-80 (p80), common in ultra-processed foods and pharmaceuticals, has raised concerns about gut health. RNA sequencing on intestinal epithelial cells (IECs) exposed to p80 revealed increased expression of ferroptosis-associated genes and disruption of lipid metabolism pathways further demonstrated by mitochondrial dysfunction, including altered membrane potential and architecture, and accumulation of reactive oxygen species, iron, lipid peroxidation, and lipid droplet formation. Lipidomic profiling identified significant alterations in triglyceride species and elevated pro-ferroptotic polyunsaturated fatty acids. These data indicate that p80 disrupts lipid homeostasis in IECs and triggers ferroptotic cell death, mechanisms potentially contributing to the increased incidence of chronic conditions like inflammatory bowel disease and metabolic syndrome. The study highlights critical implications for public health, emphasizing the need for reassessment of emulsifier safety standards while balancing needs with consumer safety.

Careful balance of the redox status of the embryo and reduction of oxidative stress is crucial in early development. Here we show that the culture of preimplantation mouse embryos in the conditionally non-essential amino acid L-proline (Pro) increases the intracellular concentration of the potent antioxidant glutathione as shown by staining of 2-cell, 4-cell and 8-cell embryos with tetrafluoroterephthalonitrile (4F-2CN). Further, liquid-chromatography/mass spectrometry showed increased GSH levels in all Pro-treated preimplantation stages of development compared to controls. The GSH:GSSG ratio also showed a Pro-dependent increase. Overall, our results indicate that the beneficial effect of Pro in preimplantation embryo culture is due to the reduction in oxidative stress mediated through an increase in cellular GSH concentration.

Mitochondria are essential for metabolic homeostasis and neuronal function, extending beyond ATP production to roles in cell signaling, inflammation, and stress responses. Mitochondrial dysfunction, marked by abnormal morphology, ATP deficiency, and oxidative stress, is a key feature of aging-related diseases and neurodegenerative disorders like Parkinsons. Given the importance of mitochondrial homeostasis to brain function, this study aimed to determine the possible vitamin D (VD3) effects on mitochondrial susceptibility to Ca2+-induced mitochondrial permeability transition pore (mPTP), bioenergetics in brain mitochondria, and redox balance. We demonstrated that VD3 protects isolated brain mitochondria. Male rats were divided into control and VD3-treated groups. Brain mitochondria were isolated for assessments of Ca2+-induced mitochondrial swelling secondary to MPTP opening, oxygen consumption (states 3 - ADP-stimulated and state 4 - in the presence of oligomycin), and the respiratory control ratio (RCR). Oxidative stress parameters (nitrite and lipid peroxidation), superoxide dismutase (SOD) activity, and reduced glutathione (GSH) levels were also evaluated. The results revealed that VD3 treatment blocked Ca2+-induced mitochondrial swelling secondary to MPTP opening. Additionally, VD3 improved mitochondrial RCR compared to controls, in the presence of complex I (malate/glutamate) and complex II (succinate) substrates, reduced mitochondrial succinate-driven H2O2 release, and enhanced SOD activity and GSH levels. These changes occurred in parallel with decreased nitrite and TBARS formation. These results suggest that vitamin D{square} confers mitochondrial neuroprotection, emphasizing its prospective role in maintaining neuronal homeostasis and mitigating neurodegenerative processes.

Smoking, alcohol abuse, and hypertension are - among others the potential risk factors for cardiovascular diseases. These risk factors generate oxidative stress and cause oxidative stress-induced DNA damage, resulting in cellular senescence and senescence-associated secretory phenotype (SASP). The SASP factors in feed-forward response exacerbate inflammation and cause tissue remodeling, resulting in atherosclerotic plaque formation and rupture. Colchicine was used to ameliorate oxidative stress-induced senescence in human umbilical vein endothelial cells. Oxidative stress was quantified by reactive oxygen species (ROS) assay and oxidative stress-induced DNA damage was analyzed by 8-OHDG immunofluorescence staining. Endothelial cell senescence was visualized by {beta}-gal staining. The relative mRNA expression was quantified by qPCR and protein analysis was performed by Western blot. Colchicine inhibited ROS generation and mitigated oxidative stress-induced DNA damage. It dampened oxidative stress-induced endothelial cell senescence and improved the expression of DNA repair protein KU80 and aging marker Lamin B1. The drug attenuated the expression of senescence marker P21 at mRNA and protein levels. The pathway analysis showed that colchicine inhibited NF-{kappa}B and MAPKs pathways and subdued mTOR activation. Colchicine also attenuated mRNA expression of interleukin (IL)-1{beta}, IL-6, IL-8 MCP-1, ICAM-1, and E-selectin. Furthermore, colchicine reduced the mRNA and protein expression of matrix metalloproteinase (MMP-2). In summary, colchicine blocked oxidative stress-induced senescence and SASP by inhibiting the activation of NF-{kappa}B and MAPKs pathways.