Pub Date : 2026-02-01DOI: 10.1016/j.freeradbiomed.2026.01.061
Rasheed A Abdulraheem, Ammar U Danazumi, Philipp Nitschke, Luke Gray Whiley, Abdulrahman Ibrahim Tudu, Ranil Coorey, Zhoyu Li, Prashant Bharadwaj, Vijay Jayasena, Stuart K Johnson, Ralph N Martins, W M A D Binosha Fernando
The accumulation of amyloid-beta (Aβ42) plaques is a hallmark of Alzheimer's disease (AD), which currently have no cure. The 3-deoxyanthocyanidins (3-DXA) and their derivatives represent a more stable class of polyphenols, identified in sorghum grains at uniquely high concentrations. Although 3-DXA exhibit strong potential to modulate protein aggregation processes, their effects on AD pathology remain largely unexplored. In this study, we investigated the inhibitory effects of three 3-DXA derivatives apigeninidin chloride (AC), luteolinidin chloride (LC), and 7-methoxy apigeninidin (7-MAC) on Aβ42 aggregation and associated neurotoxicity. Thioflavin T (ThT) fluorescence assay was employed to assess alterations in Aβ42 aggregation, while circular dichroism and nuclear magnetic resonance spectroscopy were used to evaluate changes in secondary structure. The neuroprotective effects of the 3-DXA derivatives were further examined in MC-65 cells under Aβ-induced toxicity. Additionally, generalized replica exchange with solute tempering based molecular dynamics simulations was conducted to explore the effects of AC and LC on Aβ42 dimer stability and β-sheet disruption. Our findings demonstrate that AC, LC, and 7-MAC significantly reduced Aβ42 aggregation by up to 88%, with AC and LC showing particularly strong disruption of β-sheet structures. All three compounds significantly rescued MC-65 cells from Aβ42-induced toxicity (62-77%) and enhanced mitochondrial activity. Molecular dynamics simulations analyses revealed that AC and LC disrupted hydrophobic interactions within Aβ42 dimers, contributing to destabilisation of neurotoxic aggregates. Overall, AC and LC exhibited strong multitarget activity against AD pathology by inhibiting Aβ42 aggregation, restoring intracellular energy balance, and disrupting key neurotoxic structural motifs.
{"title":"3-Deoxyanthocyanidins Inhibit β-Amyloid Aggregation, Toxicity, and Mitochondrial Dysfunction: Evidence from MC-65 Cells and Molecular Dynamics Simulations.","authors":"Rasheed A Abdulraheem, Ammar U Danazumi, Philipp Nitschke, Luke Gray Whiley, Abdulrahman Ibrahim Tudu, Ranil Coorey, Zhoyu Li, Prashant Bharadwaj, Vijay Jayasena, Stuart K Johnson, Ralph N Martins, W M A D Binosha Fernando","doi":"10.1016/j.freeradbiomed.2026.01.061","DOIUrl":"https://doi.org/10.1016/j.freeradbiomed.2026.01.061","url":null,"abstract":"<p><p>The accumulation of amyloid-beta (Aβ<sub>42</sub>) plaques is a hallmark of Alzheimer's disease (AD), which currently have no cure. The 3-deoxyanthocyanidins (3-DXA) and their derivatives represent a more stable class of polyphenols, identified in sorghum grains at uniquely high concentrations. Although 3-DXA exhibit strong potential to modulate protein aggregation processes, their effects on AD pathology remain largely unexplored. In this study, we investigated the inhibitory effects of three 3-DXA derivatives apigeninidin chloride (AC), luteolinidin chloride (LC), and 7-methoxy apigeninidin (7-MAC) on Aβ<sub>42</sub> aggregation and associated neurotoxicity. Thioflavin T (ThT) fluorescence assay was employed to assess alterations in Aβ<sub>42</sub> aggregation, while circular dichroism and nuclear magnetic resonance spectroscopy were used to evaluate changes in secondary structure. The neuroprotective effects of the 3-DXA derivatives were further examined in MC-65 cells under Aβ-induced toxicity. Additionally, generalized replica exchange with solute tempering based molecular dynamics simulations was conducted to explore the effects of AC and LC on Aβ<sub>42</sub> dimer stability and β-sheet disruption. Our findings demonstrate that AC, LC, and 7-MAC significantly reduced Aβ<sub>42</sub> aggregation by up to 88%, with AC and LC showing particularly strong disruption of β-sheet structures. All three compounds significantly rescued MC-65 cells from Aβ<sub>42</sub>-induced toxicity (62-77%) and enhanced mitochondrial activity. Molecular dynamics simulations analyses revealed that AC and LC disrupted hydrophobic interactions within Aβ<sub>42</sub> dimers, contributing to destabilisation of neurotoxic aggregates. Overall, AC and LC exhibited strong multitarget activity against AD pathology by inhibiting Aβ42 aggregation, restoring intracellular energy balance, and disrupting key neurotoxic structural motifs.</p>","PeriodicalId":12407,"journal":{"name":"Free Radical Biology and Medicine","volume":" ","pages":""},"PeriodicalIF":8.2,"publicationDate":"2026-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146112340","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2026-01-31DOI: 10.1016/j.freeradbiomed.2026.01.040
Xin Su, Teng Fan, Zeyu Liu, Yuanyuan Huang, Jun Kan, Chuwen Liang, Yuwen Chen, Zhangqi Cao, Shuangli Zhu, Sijia Li, Kai Fu, Can Pan, Fang Wang, Bei Zhang, Liwu Fu
Introduction: Doxorubicin (DOX) is a widely used chemotherapeutic agent, but its clinical application is limited by dose-dependent cardiotoxicity. Currently, there are no effective strategies to prevent or reverse DOX-mediated myocardial injury, highlighting the urgent need for novel therapeutic approaches.
Objectives: In this study, the cardioprotective effects of crocin, a natural compound derived from Crocus sativus, were investigated in the context of DOX-mediated cardiotoxicity.
Methods: Cardiac function, mitochondrial morphology, ROS production, and ATP content were evaluated in both in vitro and in vivo models of DOX-mediated cardiotoxicity. RNA sequencing was performed to identify key regulatory pathways affected by crocin. Mitophagy-related mechanisms were investigated through molecular and cellular assays, including immunofluorescence and Western blot analysis of PTEN-induced kinase 1 (PINK1)-associated signaling. PINK1 knockdown and mitophagy inhibition were performed to assess the impact on the cardioprotective effects of crocin.
Results: Crocin treatment preserved cardiac function and mitigated DOX-mediated myocardial injury in both in vitro and in vivo models, as evidenced by restored left ventricular ejection fraction, reduced mitochondrial ROS accumulation, restoration of ATP production, and improved mitochondrial morphology. Transcriptomic analysis revealed that crocin upregulated PINK1 expression, a key initiator of mitophagy. Functional assays further confirmed that crocin restored mitophagy activity suppressed by DOX exposure. The cardioprotective effects of crocin were abolished upon PINK1 knockdown or mitophagy inhibitor, highlighting the essential role of PINK1-dependent mitophagy in mediating crocin's effects.
Conclusions: Crocin protects against doxorubicin-induced cardiotoxicity by activating PINK1-mediated mitophagy and maintaining mitochondrial homeostasis. These findings highlight crocin as a potential therapeutic agent for mitigating DOX-mediated cardiotoxicity.
{"title":"Crocin alleviates doxorubicin-mediated cardiotoxicity by activating PINK1-dependent cardiomyocyte mitophagy.","authors":"Xin Su, Teng Fan, Zeyu Liu, Yuanyuan Huang, Jun Kan, Chuwen Liang, Yuwen Chen, Zhangqi Cao, Shuangli Zhu, Sijia Li, Kai Fu, Can Pan, Fang Wang, Bei Zhang, Liwu Fu","doi":"10.1016/j.freeradbiomed.2026.01.040","DOIUrl":"10.1016/j.freeradbiomed.2026.01.040","url":null,"abstract":"<p><strong>Introduction: </strong>Doxorubicin (DOX) is a widely used chemotherapeutic agent, but its clinical application is limited by dose-dependent cardiotoxicity. Currently, there are no effective strategies to prevent or reverse DOX-mediated myocardial injury, highlighting the urgent need for novel therapeutic approaches.</p><p><strong>Objectives: </strong>In this study, the cardioprotective effects of crocin, a natural compound derived from Crocus sativus, were investigated in the context of DOX-mediated cardiotoxicity.</p><p><strong>Methods: </strong>Cardiac function, mitochondrial morphology, ROS production, and ATP content were evaluated in both in vitro and in vivo models of DOX-mediated cardiotoxicity. RNA sequencing was performed to identify key regulatory pathways affected by crocin. Mitophagy-related mechanisms were investigated through molecular and cellular assays, including immunofluorescence and Western blot analysis of PTEN-induced kinase 1 (PINK1)-associated signaling. PINK1 knockdown and mitophagy inhibition were performed to assess the impact on the cardioprotective effects of crocin.</p><p><strong>Results: </strong>Crocin treatment preserved cardiac function and mitigated DOX-mediated myocardial injury in both in vitro and in vivo models, as evidenced by restored left ventricular ejection fraction, reduced mitochondrial ROS accumulation, restoration of ATP production, and improved mitochondrial morphology. Transcriptomic analysis revealed that crocin upregulated PINK1 expression, a key initiator of mitophagy. Functional assays further confirmed that crocin restored mitophagy activity suppressed by DOX exposure. The cardioprotective effects of crocin were abolished upon PINK1 knockdown or mitophagy inhibitor, highlighting the essential role of PINK1-dependent mitophagy in mediating crocin's effects.</p><p><strong>Conclusions: </strong>Crocin protects against doxorubicin-induced cardiotoxicity by activating PINK1-mediated mitophagy and maintaining mitochondrial homeostasis. These findings highlight crocin as a potential therapeutic agent for mitigating DOX-mediated cardiotoxicity.</p>","PeriodicalId":12407,"journal":{"name":"Free Radical Biology and Medicine","volume":" ","pages":"668-681"},"PeriodicalIF":8.2,"publicationDate":"2026-01-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146104161","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2026-01-30DOI: 10.1016/j.freeradbiomed.2026.01.041
Jie Zheng, Yu An, Xin Zhang, Zhaoming Cao, Guangyi Xu, Jing Li, Jingya Wang, Li Chen, Yanhui Lu
Background: Diabetic cognitive impairment (DCI) is an increasingly recognized complication of type 2 diabetes mellitus (T2DM) with limited effective therapies. Short-chain fatty acids (SCFAs) have been implicated in metabolic regulation and neuronal health, yet comparisons of acetate, propionate, butyrate, and their mixture are limited, and the mechanisms underlying neuroprotection in DCI remain insufficiently clarified.
Methods: Ninety participants (healthy controls, T2DM, and DCI groups) were assessed for serum SCFA levels and cognitive performance using the Montreal Cognitive Assessment (MoCA). In parallel, a DCI mouse model established by a 24-week high-fat diet received 8-week supplementation with acetate, propionate, butyrate, or a mixture of the three. Glucolipid metabolism, spatial learning and memory, hippocampal neuronal damage, neuroinflammation, and mitophagy were evaluated. Based on consistency across the clinical and animal datasets, acetate was selected for mitophagy-focused mechanistic experiments, and pathway dependence was examined by co-administration of the autophagy inhibitor 3-methyladenine (3-MA).
Results: Clinically, serum acetate, propionate, and butyrate were lower in T2DM and DCI than in healthy controls; only acetate showed a further significant reduction in DCI compared with T2DM. All three SCFAs were positively associated with MoCA score and inversely associated with fasting blood glucose, whereas acetate additionally showed inverse associations with lipid parameters. In mice, SCFA supplementation alleviated metabolic dysfunction, spatial learning and memory, neuronal loss, and neuroinflammation, with acetate generally producing more consistent and numerically greater improvements across these endpoints. Mechanistically, acetate enhanced hippocampal mitophagy by restoring LC3-TOMM20 colocalization and activating the PINK1/Parkin pathway. Importantly, 3-MA partially attenuated these benefits, indicating a mitophagy-dependent mechanism.
Conclusions: These integrated clinical and experimental data support a "SCFAs-mitophagy-neuroinflammation" axis linking systemic metabolism to neuronal vulnerability in DCI, and identify acetate as a promising SCFA that may enhance neuronal resilience through mitophagy activation.
{"title":"Acetate ameliorates glucolipid metabolic dysregulation and neuroinflammation in diabetic cognitive impairment via enhanced mitophagy.","authors":"Jie Zheng, Yu An, Xin Zhang, Zhaoming Cao, Guangyi Xu, Jing Li, Jingya Wang, Li Chen, Yanhui Lu","doi":"10.1016/j.freeradbiomed.2026.01.041","DOIUrl":"10.1016/j.freeradbiomed.2026.01.041","url":null,"abstract":"<p><strong>Background: </strong>Diabetic cognitive impairment (DCI) is an increasingly recognized complication of type 2 diabetes mellitus (T2DM) with limited effective therapies. Short-chain fatty acids (SCFAs) have been implicated in metabolic regulation and neuronal health, yet comparisons of acetate, propionate, butyrate, and their mixture are limited, and the mechanisms underlying neuroprotection in DCI remain insufficiently clarified.</p><p><strong>Methods: </strong>Ninety participants (healthy controls, T2DM, and DCI groups) were assessed for serum SCFA levels and cognitive performance using the Montreal Cognitive Assessment (MoCA). In parallel, a DCI mouse model established by a 24-week high-fat diet received 8-week supplementation with acetate, propionate, butyrate, or a mixture of the three. Glucolipid metabolism, spatial learning and memory, hippocampal neuronal damage, neuroinflammation, and mitophagy were evaluated. Based on consistency across the clinical and animal datasets, acetate was selected for mitophagy-focused mechanistic experiments, and pathway dependence was examined by co-administration of the autophagy inhibitor 3-methyladenine (3-MA).</p><p><strong>Results: </strong>Clinically, serum acetate, propionate, and butyrate were lower in T2DM and DCI than in healthy controls; only acetate showed a further significant reduction in DCI compared with T2DM. All three SCFAs were positively associated with MoCA score and inversely associated with fasting blood glucose, whereas acetate additionally showed inverse associations with lipid parameters. In mice, SCFA supplementation alleviated metabolic dysfunction, spatial learning and memory, neuronal loss, and neuroinflammation, with acetate generally producing more consistent and numerically greater improvements across these endpoints. Mechanistically, acetate enhanced hippocampal mitophagy by restoring LC3-TOMM20 colocalization and activating the PINK1/Parkin pathway. Importantly, 3-MA partially attenuated these benefits, indicating a mitophagy-dependent mechanism.</p><p><strong>Conclusions: </strong>These integrated clinical and experimental data support a \"SCFAs-mitophagy-neuroinflammation\" axis linking systemic metabolism to neuronal vulnerability in DCI, and identify acetate as a promising SCFA that may enhance neuronal resilience through mitophagy activation.</p>","PeriodicalId":12407,"journal":{"name":"Free Radical Biology and Medicine","volume":" ","pages":"580-597"},"PeriodicalIF":8.2,"publicationDate":"2026-01-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146100175","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Mycobacterium tuberculosis (Mtb) Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) is indispensable for glycolysis, it also performs several critical non-metabolic functions. In the present study, we demonstrate that CRISPRi silencing of GAPDH inhibited enzyme activity and iron acquisition via human transferrin (Tf)/lactoferrin (Lf). GAPDH silencing also enhanced reactive oxygen species (ROS) and ROS induced damage suggesting its role as a redox sensor. We then examined the impact of GAPDH inhibition in Mtb using small molecule inhibitors. Vitamin C (VC) was selected considering its potent bactericidal effects against Mtb and its inhibition of human GAPDH resulting in its efficacy against cancer cells. The GAPDH inhibitors Ethyl bromopyruvate (EBP) and Koningic acid (KA) are anti-cancer agents that target the glycolytic activity of GAPDH. In contrast, TCH346 was identified as a neuroprotective agent, wherein it targets the non-metabolic function of GAPDH induced apoptotic signalling. The effects of inhibitors, alone or in combination with VC mirrored the cellular effects of GAPDH silencing, resulting in significant anti-bacterial activity. VC induced iron mobilization which coupled with GAPDH inhibitors induced a veritable "double whammy" resulting in massive increase in ROS and downstream effects. The efficacy of these treatments was assessed in a murine model, confirming that VC augmented the potent anti-tubercular activity induced by EBP and TCH346. Overall, this study identifies the crucial function of Mtb GAPDH as a redox sensor and highlights the potential of targeting its pleiotropic cellular functions towards drug discovery. In addition, the efficacy of TCH346 provides an opportunity of drug-repurposing as a strategy for therapy.
{"title":"Targeting Mycobacterium tuberculosis GAPDH elicits potent bactericidal responses by dysregulating enzyme activity, redox dynamics and iron acquisition.","authors":"Zahid Gani, Mohammad Naiyaz Ahmad, Anurag Sindhu, Ajay Kumar, Anjali Kumari, Mohmmad Imran, Pradip Malik, Asmita Dhiman, Vinay Kumar Yadav, Gaddam Laxmi Priya, Gattadi Sravani, Nisheeth Agarwal, Rajender Kumar, Prabha Garg, Arunava Dasgupta, Sidharth Chopra, Manoj Raje, Chaaya Iyengar Raje","doi":"10.1016/j.freeradbiomed.2026.01.044","DOIUrl":"10.1016/j.freeradbiomed.2026.01.044","url":null,"abstract":"<p><p>Mycobacterium tuberculosis (Mtb) Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) is indispensable for glycolysis, it also performs several critical non-metabolic functions. In the present study, we demonstrate that CRISPRi silencing of GAPDH inhibited enzyme activity and iron acquisition via human transferrin (Tf)/lactoferrin (Lf). GAPDH silencing also enhanced reactive oxygen species (ROS) and ROS induced damage suggesting its role as a redox sensor. We then examined the impact of GAPDH inhibition in Mtb using small molecule inhibitors. Vitamin C (VC) was selected considering its potent bactericidal effects against Mtb and its inhibition of human GAPDH resulting in its efficacy against cancer cells. The GAPDH inhibitors Ethyl bromopyruvate (EBP) and Koningic acid (KA) are anti-cancer agents that target the glycolytic activity of GAPDH. In contrast, TCH346 was identified as a neuroprotective agent, wherein it targets the non-metabolic function of GAPDH induced apoptotic signalling. The effects of inhibitors, alone or in combination with VC mirrored the cellular effects of GAPDH silencing, resulting in significant anti-bacterial activity. VC induced iron mobilization which coupled with GAPDH inhibitors induced a veritable \"double whammy\" resulting in massive increase in ROS and downstream effects. The efficacy of these treatments was assessed in a murine model, confirming that VC augmented the potent anti-tubercular activity induced by EBP and TCH346. Overall, this study identifies the crucial function of Mtb GAPDH as a redox sensor and highlights the potential of targeting its pleiotropic cellular functions towards drug discovery. In addition, the efficacy of TCH346 provides an opportunity of drug-repurposing as a strategy for therapy.</p>","PeriodicalId":12407,"journal":{"name":"Free Radical Biology and Medicine","volume":" ","pages":"54-70"},"PeriodicalIF":8.2,"publicationDate":"2026-01-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146100139","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Introduction: Epidemiological studies have demonstrated higher incidence and mortality rate of nonalcoholic steatohepatitis (NASH) in the elderly population than in younger groups. However, the mechanisms underlying this age-related exacerbation remain poorly understood.
Objective: This study aimed to elucidate the specific pathways through which aging exacerbates NASH progression, using an integrated in vivo and in vitro model.
Methods: Aged (18-month-old) and young (6-week-old) mice were fed a high-fat diet (HFD) for 16 weeks to induce NASH. A senescence-associated cellular model of NASH was established by co-treating murine hepatocyte AML-12 with H2O2 and free fatty acid (FFA). Gene expression profiling of liver tissue was performed using RNA sequencing to identify molecular signatures. Interventions were as follows: (1) In vitro, BMAL1 overexpression plasmids were transfected into AML-12 cells, followed by treatment with 2-deoxy-D-glucose (2-DG, a glycolysis inhibitor) and 2-methoxyestradiol (2-ME2, a HIF-1α inhibitor); (2) in vivo, hepatocyte-specific BMAL1 overexpression was achieved in aged HFD-fed mice through adeno-associated virus serotype 8 (AAV8) delivery. Mechanism validation was performed using biochemical assays, Western blot, cell staining, molecular docking, and Co-IP.
Results: Aged HFD-fed mice exhibited more severe NASH phenotypes than young mice. Transcriptomic analysis identified NLRP3-related signaling and circadian rhythm pathways as central contributors to age-specific NASH pathogenesis. These mice also exhibited elevated NLRP3 inflammasome activity, enhanced glycolysis, and reduced BMAL1 expression. In senescent NASH cells, BMAL1 overexpression along with 2-DG or 2-ME2 treatment significantly downregulated NLRP3 expression and attenuated lipid accumulation, inflammation, oxidative stress, and fibrosis. Mechanistically, BMAL1 directly bound to HIF-1α, thereby suppressing glycolysis. Hepatocyte-specific BMAL1 overexpression in aged HFD-fed mice markedly inhibited glycolysis and NLRP3 activation, resulting in an improvement in NASH-related pathologies.
Conclusion: This study revealed a novel mechanism in which BMAL1 downregulation under aging and HFD conditions promotes NASH progression by binding to HIF-1α and modulating the glycolysis-NLRP3 inflammasome axis.
{"title":"BMAL1 downregulation exacerbates age-related nonalcoholic steatohepatitis by promoting NLRP3 inflammasome activation via HIF-1ɑ-mediated glycolysis.","authors":"Yujie Ren, Dongying Lv, Jiayan Chen, Wenjing Chen, Chu Chen, Lizong Zhang, Jue Tu, Keyan Zhu, Dejun Wang, Zhaowei Cai","doi":"10.1016/j.freeradbiomed.2026.01.058","DOIUrl":"10.1016/j.freeradbiomed.2026.01.058","url":null,"abstract":"<p><strong>Introduction: </strong>Epidemiological studies have demonstrated higher incidence and mortality rate of nonalcoholic steatohepatitis (NASH) in the elderly population than in younger groups. However, the mechanisms underlying this age-related exacerbation remain poorly understood.</p><p><strong>Objective: </strong>This study aimed to elucidate the specific pathways through which aging exacerbates NASH progression, using an integrated in vivo and in vitro model.</p><p><strong>Methods: </strong>Aged (18-month-old) and young (6-week-old) mice were fed a high-fat diet (HFD) for 16 weeks to induce NASH. A senescence-associated cellular model of NASH was established by co-treating murine hepatocyte AML-12 with H<sub>2</sub>O<sub>2</sub> and free fatty acid (FFA). Gene expression profiling of liver tissue was performed using RNA sequencing to identify molecular signatures. Interventions were as follows: (1) In vitro, BMAL1 overexpression plasmids were transfected into AML-12 cells, followed by treatment with 2-deoxy-D-glucose (2-DG, a glycolysis inhibitor) and 2-methoxyestradiol (2-ME2, a HIF-1α inhibitor); (2) in vivo, hepatocyte-specific BMAL1 overexpression was achieved in aged HFD-fed mice through adeno-associated virus serotype 8 (AAV8) delivery. Mechanism validation was performed using biochemical assays, Western blot, cell staining, molecular docking, and Co-IP.</p><p><strong>Results: </strong>Aged HFD-fed mice exhibited more severe NASH phenotypes than young mice. Transcriptomic analysis identified NLRP3-related signaling and circadian rhythm pathways as central contributors to age-specific NASH pathogenesis. These mice also exhibited elevated NLRP3 inflammasome activity, enhanced glycolysis, and reduced BMAL1 expression. In senescent NASH cells, BMAL1 overexpression along with 2-DG or 2-ME2 treatment significantly downregulated NLRP3 expression and attenuated lipid accumulation, inflammation, oxidative stress, and fibrosis. Mechanistically, BMAL1 directly bound to HIF-1α, thereby suppressing glycolysis. Hepatocyte-specific BMAL1 overexpression in aged HFD-fed mice markedly inhibited glycolysis and NLRP3 activation, resulting in an improvement in NASH-related pathologies.</p><p><strong>Conclusion: </strong>This study revealed a novel mechanism in which BMAL1 downregulation under aging and HFD conditions promotes NASH progression by binding to HIF-1α and modulating the glycolysis-NLRP3 inflammasome axis.</p>","PeriodicalId":12407,"journal":{"name":"Free Radical Biology and Medicine","volume":" ","pages":"562-579"},"PeriodicalIF":8.2,"publicationDate":"2026-01-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146100151","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Extrusion of damaged mitochondria is emerging as a trigger of innate immune activation. Parkinson's disease (PD), characterized by profound mitochondrial dysfunction, may involve similar mechanisms. Here, we report that dopaminergic neurons release damaged mitochondria into the extracellular space in the 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) mouse model of PD. These neuron-derived mitochondria were subsequently engulfed by glial cells, eliciting robust inflammatory responses. Autophagy inhibition did not affect mitochondrial release, indicating a non-canonical extrusion pathway. Upon mitochondrial damage, Rab27a and Rab27b translocated to the outer mitochondrial membrane, mediating mitochondrial export from dopaminergic neurons. Conditional Rab27 knockdown in dopaminergic neurons reduced extracellular mitochondrial accumulation, microglial activation, antiviral signaling, and dopaminergic neurodegeneration. Together, these findings identify Rab27-dependent mitochondrial extrusion as a critical mechanism coupling dopaminergic neuronal injury to neuroinflammation and neurodegeneration in PD.
{"title":"Rab27-dependent mitochondrial extrusion from dopaminergic neurons drives neuroinflammation and neurodegeneration in the MPTP mouse model of Parkinson's disease.","authors":"Yingqi Xu, Junyu Li, Shanshan Ma, Ting Yang, Ziyue Shen, Mingtao Li, Qiaoying Huang","doi":"10.1016/j.freeradbiomed.2026.01.053","DOIUrl":"https://doi.org/10.1016/j.freeradbiomed.2026.01.053","url":null,"abstract":"<p><p>Extrusion of damaged mitochondria is emerging as a trigger of innate immune activation. Parkinson's disease (PD), characterized by profound mitochondrial dysfunction, may involve similar mechanisms. Here, we report that dopaminergic neurons release damaged mitochondria into the extracellular space in the 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) mouse model of PD. These neuron-derived mitochondria were subsequently engulfed by glial cells, eliciting robust inflammatory responses. Autophagy inhibition did not affect mitochondrial release, indicating a non-canonical extrusion pathway. Upon mitochondrial damage, Rab27a and Rab27b translocated to the outer mitochondrial membrane, mediating mitochondrial export from dopaminergic neurons. Conditional Rab27 knockdown in dopaminergic neurons reduced extracellular mitochondrial accumulation, microglial activation, antiviral signaling, and dopaminergic neurodegeneration. Together, these findings identify Rab27-dependent mitochondrial extrusion as a critical mechanism coupling dopaminergic neuronal injury to neuroinflammation and neurodegeneration in PD.</p>","PeriodicalId":12407,"journal":{"name":"Free Radical Biology and Medicine","volume":" ","pages":""},"PeriodicalIF":8.2,"publicationDate":"2026-01-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146096920","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Myocardial infarction (MI) stands as a leading contributor to global cardiovascular morbidity and mortality, defined by ischemic myocardial cell death and subsequent impairment of cardiac function. The tripartite motif (TRIM) protein family has been shown to regulate myocardial ischemia-reperfusion injury. As a key member of the TRIM protein family, tripartite motif-containing protein 28 (TRIM28) exhibits dysregulated expression in the heart during MI yet its pathophysiological role remains to be fully elucidated. This study aimed to investigate the functional roles and underlying mechanisms of TRIM28 in MI. We observed a significant upregulation of TRIM28 in ischemic myocardium and hypoxic cardiomyocytes. Genetic knockout of TRIM28 ameliorated cardiac function and attenuated apoptosis in MI mice, whereas its overexpression exacerbated contractile dysfunction, and promoted cardiomyocyte apoptosis and mitochondrial injury. Mechanistically, TRIM28 directly interacts with activating transcription factor 5 (ATF5) and suppresses its SUMOylation, thereby enhancing the ubiquitin-mediated degradation of ATF5, inhibiting the mitochondrial unfolded protein response (UPRmt), and ultimately culminating in increased apoptosis. Via molecular docking, we identified a TRIM28-targeting compound, Oolonghomobisflavan B (OFB), which attenuated post-MI apoptosis and facilitated cardiac function recovery. Collectively, these findings demonstrate that TRIM28 acts as a critical regulator of MI progression, and OFB holds therapeutic potential as a candidate drug.
心肌梗死(MI)是全球心血管发病率和死亡率的主要原因,其定义为缺血性心肌细胞死亡和随后的心功能损害。tripartite motif (TRIM)蛋白家族已被证明可调节心肌缺血再灌注损伤。TRIM28 (tripartite motif-containing protein 28, TRIM28)作为TRIM蛋白家族的关键成员,在心肌梗死时表现出异常表达,但其病理生理作用尚不清楚。本研究旨在探讨TRIM28在心肌梗死中的功能作用及其机制。我们观察到TRIM28在缺血心肌和缺氧心肌细胞中显著上调。基因敲除TRIM28可改善心肌梗死小鼠的心功能,减轻心肌细胞凋亡,而其过表达可加重心肌收缩功能障碍,促进心肌细胞凋亡和线粒体损伤。在机制上,TRIM28直接与活化转录因子5 (ATF5)相互作用,抑制其SUMOylation,从而增强泛素介导的ATF5降解,抑制线粒体未折叠蛋白反应(UPRmt),最终导致细胞凋亡增加。通过分子对接,我们发现了一种靶向trim28的化合物Oolonghomobisflavan B (OFB),它可以减轻心肌梗死后的细胞凋亡,促进心功能恢复。总的来说,这些发现表明TRIM28是心肌梗死进展的关键调节因子,OFB作为候选药物具有治疗潜力。
{"title":"TRIM28 aggravates myocardial infarction-induced cardiomyocyte apoptosis through regulating the stability of ATF5 via ubiquitination and SUMOylation.","authors":"Yanying Wang, Mingyu Yang, Junting Ren, Wei Liu, Han Sun, Guangze Wang, Siyu Wang, Ying Zhang, Haodong Li, Dongping Liu, Mengmeng Li, Yanwei Zhang, Hao Wang, Xuewen Yang, Xiyang Zhang, Yuhan Liu, Lei Jiao, Lihua Sun, Lina Xuan, Xuelian Li, Shasha Fan, Manyu Gong, Ying Zhang","doi":"10.1016/j.freeradbiomed.2026.01.055","DOIUrl":"10.1016/j.freeradbiomed.2026.01.055","url":null,"abstract":"<p><p>Myocardial infarction (MI) stands as a leading contributor to global cardiovascular morbidity and mortality, defined by ischemic myocardial cell death and subsequent impairment of cardiac function. The tripartite motif (TRIM) protein family has been shown to regulate myocardial ischemia-reperfusion injury. As a key member of the TRIM protein family, tripartite motif-containing protein 28 (TRIM28) exhibits dysregulated expression in the heart during MI yet its pathophysiological role remains to be fully elucidated. This study aimed to investigate the functional roles and underlying mechanisms of TRIM28 in MI. We observed a significant upregulation of TRIM28 in ischemic myocardium and hypoxic cardiomyocytes. Genetic knockout of TRIM28 ameliorated cardiac function and attenuated apoptosis in MI mice, whereas its overexpression exacerbated contractile dysfunction, and promoted cardiomyocyte apoptosis and mitochondrial injury. Mechanistically, TRIM28 directly interacts with activating transcription factor 5 (ATF5) and suppresses its SUMOylation, thereby enhancing the ubiquitin-mediated degradation of ATF5, inhibiting the mitochondrial unfolded protein response (UPRmt), and ultimately culminating in increased apoptosis. Via molecular docking, we identified a TRIM28-targeting compound, Oolonghomobisflavan B (OFB), which attenuated post-MI apoptosis and facilitated cardiac function recovery. Collectively, these findings demonstrate that TRIM28 acts as a critical regulator of MI progression, and OFB holds therapeutic potential as a candidate drug.</p>","PeriodicalId":12407,"journal":{"name":"Free Radical Biology and Medicine","volume":" ","pages":"598-613"},"PeriodicalIF":8.2,"publicationDate":"2026-01-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146096842","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Metformin (Met), a first-line therapeutic agent for type 2 diabetes, has been widely recognized for its antifibrotic properties in various pathological conditions. However, its effects on hypertrophic scars (HS) and the underlying mechanisms remain insufficiently explored. The present study aimed to elucidate the role of metformin in HS and to investigate its associated molecular mechanisms. Both in vitro and in vivo experiments demonstrated that metformin markedly inhibited the proliferation, migration, and collagen deposition of hypertrophic scar fibroblasts (HSFs), and alleviated HS formation in a rabbit ear model. Mechanistic investigations further revealed that these effects were closely associated with the downregulation of ribonucleotide reductase regulatory subunit M2 (RRM2). Notably, reduced RRM2 expression suppressed the production of glutathione synthetase (GSS), thereby impairing glutathione (GSH) synthesis. This, in turn, indirectly downregulated glutathione peroxidase 4 (GPX4), leading to the intracellular accumulation of peroxides and triggering ferroptosis in vivo and in vitro. Collectively, these findings suggest that metformin may attenuate HS fibrosis by inducing HSFs ferroptosis through the RRM2/GSS/GPX4 signaling axis. This study not only expands the potential clinical application of metformin in the treatment of skin fibrosis but also provides a theoretical foundation for the development of novel anti-scar therapeutics.
{"title":"Metformin targets RRM2/GSS/GPX4 axis to induce fibroblast ferroptosis: A foreground strategy against hypertrophic scarring.","authors":"Ziqing Chen, Xing Li, Jialei Zhong, Guochang Chen, Dinghong Min, Jiawen Fan, Jinwei Shang, Gehua Zhu, Peng Hua, Mingzhuo Liu, Guanghua Guo","doi":"10.1016/j.freeradbiomed.2026.01.056","DOIUrl":"10.1016/j.freeradbiomed.2026.01.056","url":null,"abstract":"<p><p>Metformin (Met), a first-line therapeutic agent for type 2 diabetes, has been widely recognized for its antifibrotic properties in various pathological conditions. However, its effects on hypertrophic scars (HS) and the underlying mechanisms remain insufficiently explored. The present study aimed to elucidate the role of metformin in HS and to investigate its associated molecular mechanisms. Both in vitro and in vivo experiments demonstrated that metformin markedly inhibited the proliferation, migration, and collagen deposition of hypertrophic scar fibroblasts (HSFs), and alleviated HS formation in a rabbit ear model. Mechanistic investigations further revealed that these effects were closely associated with the downregulation of ribonucleotide reductase regulatory subunit M2 (RRM2). Notably, reduced RRM2 expression suppressed the production of glutathione synthetase (GSS), thereby impairing glutathione (GSH) synthesis. This, in turn, indirectly downregulated glutathione peroxidase 4 (GPX4), leading to the intracellular accumulation of peroxides and triggering ferroptosis in vivo and in vitro. Collectively, these findings suggest that metformin may attenuate HS fibrosis by inducing HSFs ferroptosis through the RRM2/GSS/GPX4 signaling axis. This study not only expands the potential clinical application of metformin in the treatment of skin fibrosis but also provides a theoretical foundation for the development of novel anti-scar therapeutics.</p>","PeriodicalId":12407,"journal":{"name":"Free Radical Biology and Medicine","volume":" ","pages":"489-504"},"PeriodicalIF":8.2,"publicationDate":"2026-01-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146097078","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2026-01-29DOI: 10.1016/j.freeradbiomed.2026.01.036
Lihui Deng , Boyang Wang , Haipeng Jie , Meitong Liu , Luyao Yu , Shuzhen Wu , Lanlan Wang , Shengnan Li , Xiaohui Hu , Yalin Yu , Guohua Song , Bo Dong
Background
Diabetic cardiomyopathy (DCM) is a major complication of diabetes mellitus, leading to significant mortality. The (Pro)renin Receptor (PRR) is implicated in cardiovascular pathology, but its specific role in regulating mitochondrial quality control and cellular senescence in the context of DCM remains poorly understood. This study aimed to elucidate the mechanism by which PRR contributes to myocardial injury in DCM.
Methods
DCM was induced in mice using a high-fat diet combined with streptozotocin injection. The function of PRR was investigated in vivo and in high-glucose (HG)-stimulated neonatal rat cardiomyocytes (NRCMs) in vitro using adenoviral vectors for overexpression and knockdown. Cardiac function, myocardial remodeling (fibrosis, hypertrophy), mitophagy, and senescence were assessed using echocardiography, histological and immunofluorescence staining, Western blot, and RT-qPCR. RNA-sequencing was employed to identify downstream targets of PRR, and the protein-protein interaction was validated by co-immunoprecipitation and pull-down assays.
Results
PRR expression was significantly upregulated in the myocardium of DCM mice and in HG-treated NRCMs. Overexpression of PRR exacerbated cardiac dysfunction, myocardial fibrosis, and hypertrophy, which was associated with impaired mitophagy and increased cellular senescence. Conversely, genetic knockdown of PRR ameliorated these pathological changes. Mechanistically, PRR was found to physically interact with and suppress kinase activity of Leucine-rich repeat kinase 2 (LRRK2). Silencing LRRK2 abolished the protective effects of PRR knockdown, confirming that LRRK2 is a critical downstream mediator of PRR's detrimental effects.
Conclusions
PRR exacerbates diabetic cardiomyopathy by suppressing LRRK2, leading to impaired mitophagy and accelerated cellular senescence. The PRR/LRRK2 axis may be a potentially promising and novel therapeutic paradigm for treating DCM, and targeting PRR may represent a possibly promising therapeutic strategy.
{"title":"(Pro)renin receptor (PRR) exacerbates diabetic cardiomyopathy by suppressing LRRK2-Mediated mitophagy and promoting senescence","authors":"Lihui Deng , Boyang Wang , Haipeng Jie , Meitong Liu , Luyao Yu , Shuzhen Wu , Lanlan Wang , Shengnan Li , Xiaohui Hu , Yalin Yu , Guohua Song , Bo Dong","doi":"10.1016/j.freeradbiomed.2026.01.036","DOIUrl":"10.1016/j.freeradbiomed.2026.01.036","url":null,"abstract":"<div><h3>Background</h3><div>Diabetic cardiomyopathy (DCM) is a major complication of diabetes mellitus, leading to significant mortality. The (Pro)renin Receptor (PRR) is implicated in cardiovascular pathology, but its specific role in regulating mitochondrial quality control and cellular senescence in the context of DCM remains poorly understood. This study aimed to elucidate the mechanism by which PRR contributes to myocardial injury in DCM.</div></div><div><h3>Methods</h3><div>DCM was induced in mice using a high-fat diet combined with streptozotocin injection. The function of PRR was investigated in vivo and in high-glucose (HG)-stimulated neonatal rat cardiomyocytes (NRCMs) in vitro using adenoviral vectors for overexpression and knockdown. Cardiac function, myocardial remodeling (fibrosis, hypertrophy), mitophagy, and senescence were assessed using echocardiography, histological and immunofluorescence staining, Western blot, and RT-qPCR. RNA-sequencing was employed to identify downstream targets of PRR, and the protein-protein interaction was validated by co-immunoprecipitation and pull-down assays.</div></div><div><h3>Results</h3><div>PRR expression was significantly upregulated in the myocardium of DCM mice and in HG-treated NRCMs. Overexpression of PRR exacerbated cardiac dysfunction, myocardial fibrosis, and hypertrophy, which was associated with impaired mitophagy and increased cellular senescence. Conversely, genetic knockdown of PRR ameliorated these pathological changes. Mechanistically, PRR was found to physically interact with and suppress kinase activity of Leucine-rich repeat kinase 2 (LRRK2). Silencing LRRK2 abolished the protective effects of PRR knockdown, confirming that LRRK2 is a critical downstream mediator of PRR's detrimental effects.</div></div><div><h3>Conclusions</h3><div>PRR exacerbates diabetic cardiomyopathy by suppressing LRRK2, leading to impaired mitophagy and accelerated cellular senescence. The PRR/LRRK2 axis may be a potentially promising and novel therapeutic paradigm for treating DCM, and targeting PRR may represent a possibly promising therapeutic strategy.</div></div>","PeriodicalId":12407,"journal":{"name":"Free Radical Biology and Medicine","volume":"246 ","pages":"Pages 442-455"},"PeriodicalIF":8.2,"publicationDate":"2026-01-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146074884","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2026-01-29DOI: 10.1016/j.freeradbiomed.2026.01.047
Alessandra Pecorelli, Anna Guiotto, Alice Casoni, Marta Ruzza, Lorena Beltrami, Barbara Canepa, Fiorella Biasi, Giuseppe Poli, Giuseppe Valacchi
The cutaneous tissue is persistently exposed to environmental stressors, including a wide range of airborne pollutants. This chronic exposure often leads to a condition of oxidative stress, with the outermost layer of epidermis, the stratum corneum (SC), being especially vulnerable due to its high lipid content. Notably, approximately 40 % of SC lipids consist of cholesterol, present in both esterified and unesterified forms. The oxidative imbalance induced by environmental stressors and constantly associated with inflammatory skin diseases promotes the formation and accumulation of cholesterol oxidation products, belonging to the oxysterols' family, which are known for their potent pro-oxidant and pro-inflammatory properties. In addition, harmful oxysterols of dietary origin could reach the epidermis via the vascularized dermis, thus adding another route of exposure. 7β-Hydroxycholesterol (7βOHC) and 7-ketocholesterol (7 KC), two highly toxic oxysterols of non-enzymatic origin, have been shown to significantly downregulate proteins involved in adherens and tight junctions in the intestinal epithelium. Given the structural similarity of extracellular junction proteins across tissues, it is reasonable to expect that oxysterols may similarly disrupt the integrity of the epidermal barrier. To investigate this, supraphysiologic concentrations of 7 KC and 7βOHC were added to the medium of human keratinocytes. Immunofluorescence analysis revealed a consistent and significant reduction in the levels of Claudin-1, Zonulin-1 (ZO1), and E-cadherin, key proteins of tight and adherens junctions, respectively, in oxysterol-treated cells compared to controls. Notably, oxysterol exposure also led to a reduction of mitochondrial membrane potential and an increased mitochondrial reactive oxygen species (ROS) production. Both mitochondrial damage and the disruption of skin junctions were efficiently prevented by mitoTEMPO, a selective mitochondrial superoxide scavenger, suggesting the pro-oxidant activity of oxysterols mediates these effects in keratinocytes. Finally, experiments conducted using a 3D skin model corroborated findings observed in keratinocyte cultures, reinforcing the role of oxysterols in compromising the skin barrier integrity.
{"title":"Oxysterol-induced oxidative disruption of skin junction integrity.","authors":"Alessandra Pecorelli, Anna Guiotto, Alice Casoni, Marta Ruzza, Lorena Beltrami, Barbara Canepa, Fiorella Biasi, Giuseppe Poli, Giuseppe Valacchi","doi":"10.1016/j.freeradbiomed.2026.01.047","DOIUrl":"10.1016/j.freeradbiomed.2026.01.047","url":null,"abstract":"<p><p>The cutaneous tissue is persistently exposed to environmental stressors, including a wide range of airborne pollutants. This chronic exposure often leads to a condition of oxidative stress, with the outermost layer of epidermis, the stratum corneum (SC), being especially vulnerable due to its high lipid content. Notably, approximately 40 % of SC lipids consist of cholesterol, present in both esterified and unesterified forms. The oxidative imbalance induced by environmental stressors and constantly associated with inflammatory skin diseases promotes the formation and accumulation of cholesterol oxidation products, belonging to the oxysterols' family, which are known for their potent pro-oxidant and pro-inflammatory properties. In addition, harmful oxysterols of dietary origin could reach the epidermis via the vascularized dermis, thus adding another route of exposure. 7β-Hydroxycholesterol (7βOHC) and 7-ketocholesterol (7 KC), two highly toxic oxysterols of non-enzymatic origin, have been shown to significantly downregulate proteins involved in adherens and tight junctions in the intestinal epithelium. Given the structural similarity of extracellular junction proteins across tissues, it is reasonable to expect that oxysterols may similarly disrupt the integrity of the epidermal barrier. To investigate this, supraphysiologic concentrations of 7 KC and 7βOHC were added to the medium of human keratinocytes. Immunofluorescence analysis revealed a consistent and significant reduction in the levels of Claudin-1, Zonulin-1 (ZO1), and E-cadherin, key proteins of tight and adherens junctions, respectively, in oxysterol-treated cells compared to controls. Notably, oxysterol exposure also led to a reduction of mitochondrial membrane potential and an increased mitochondrial reactive oxygen species (ROS) production. Both mitochondrial damage and the disruption of skin junctions were efficiently prevented by mitoTEMPO, a selective mitochondrial superoxide scavenger, suggesting the pro-oxidant activity of oxysterols mediates these effects in keratinocytes. Finally, experiments conducted using a 3D skin model corroborated findings observed in keratinocyte cultures, reinforcing the role of oxysterols in compromising the skin barrier integrity.</p>","PeriodicalId":12407,"journal":{"name":"Free Radical Biology and Medicine","volume":" ","pages":"505-517"},"PeriodicalIF":8.2,"publicationDate":"2026-01-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146097116","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}