Free Radical Biology and Medicine最新文献

{"title":"Platelet COX and LOX enzymes orchestrate amyloid-β secretion via RhoA signaling: Implications for neurodegenerative diseases","authors":"A. Trostchansky ,&nbsp;M. Alarcón","doi":"10.1016/j.freeradbiomed.2025.08.060","DOIUrl":"10.1016/j.freeradbiomed.2025.08.060","url":null,"abstract":"<div><div>Arachidonic acid metabolism through cyclooxygenase (COX) and lipoxygenase (LOX) pathways is fundamental to inflammation, vascular homeostasis, and neuronal signaling. Here, we investigated the roles of platelet-expressed COX (PTGS1) and LOX (ALOX12) isoforms in amyloid-β (Aβ) secretion, a process implicated in the pathogenesis of cerebral amyloid angiopathy (CAA) and Alzheimer's disease (AD). Using an integrative approach combining bioinformatic protein–protein interaction mapping, pathway enrichment analysis, and experimental validation, we identified extensive networks linking PTGS and ALOX isoforms to cytoskeletal remodeling, mitochondrial function, and vesicle trafficking. Functional enrichment pointed to key roles for PTGS1 and ALOX12 in platelet activation and secretory processes. In vitro studies demonstrated that stimulation of human platelets with TRAP-6 triggered a robust increase in Aβ40 secretion, which was significantly attenuated by COX inhibition or blockade of RhoA, a critical regulator of cytoskeletal dynamics. These findings suggest that platelet-derived Aβ release is driven by COX/LOX-dependent signaling via RhoA. While our results support a COX/LOX-RhoA axis, we recognize that causality remains to be fully established, and the role of ALOX12 requires further experimental validation. Given the vascular deposition of Aβ40 in CAA, our results position platelets as important peripheral contributors to neurovascular amyloidosis. This study should therefore be viewed as hypothesis-generating, underscoring the therapeutic potential of targeting platelet signaling pathways to mitigate Aβ-driven vascular pathology.</div></div>","PeriodicalId":12407,"journal":{"name":"Free Radical Biology and Medicine","volume":"240 ","pages":"Pages 641-649"},"PeriodicalIF":8.2,"publicationDate":"2025-09-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145000050","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}

{"title":"Effects of aging and anti-aging dietary restriction on regulators of the [NADPH]/[NADP+] in different neural cell types and brain regions","authors":"Leah E. Jamerson,&nbsp;Patrick C. Bradshaw","doi":"10.1016/j.freeradbiomed.2025.09.001","DOIUrl":"10.1016/j.freeradbiomed.2025.09.001","url":null,"abstract":"<div><div>Dietary restriction (DR), which slows aging, increases the ratio of reduced glutathione (GSH) to oxidized glutathione disulfide (GSSG) in the brain. DR increases liver cytoplasmic [NADPH]/[NADP⁺] where much of the NADPH is generated by the folate cycle. This could also occur in astrocytes, the neural cell type with the highest folate cycle flux. Mice on a DR diet showed increased expression of folate cycle enzyme MTHFD1L in several brain regions and likely show increased astrocyte sarcosine catabolism increasing folate cycle cytoplasmic NADPH generation by ALDH1L1. Fasting also increases blood malate/pyruvate that increases tissue [NADPH]/[NADP⁺]. These events together with decreased NADPH-utilizing lipid synthesis during DR could lead to an increased brain cytoplasmic [NADPH]/[NADP⁺]. The more reduced NADP(H) pool, combined with the increased expression of brain glutathione disulfide reductase (GSR) and the decreased brain mitochondrial H₂O₂ generation, decreasing H₂O₂-induced oxidation of GSH, could lead to the increased brain GSH/GSSG. Aging also decreased the expression of mouse hippocampal NAD⁺ kinase (NADK) that was restored by DR. Studies that measure the [NADPH]/[NADP⁺], cysteine/cystine, and GSH/GSSG in different brain regions, subcellular compartments, and neural cell types, especially in astrocytes, during aging and DR are needed to establish effective targets and therapies for aging-related disorders.</div></div>","PeriodicalId":12407,"journal":{"name":"Free Radical Biology and Medicine","volume":"240 ","pages":"Pages 745-772"},"PeriodicalIF":8.2,"publicationDate":"2025-09-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144999997","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}

{"title":"Single-cell, spatial and bulk transcriptome data analysis revealed LINC00467-mediated Sertoli cell ferroptosis is a potential therapeutic target and biomarker for azoospermia","authors":"Hao Bo ,&nbsp;Fang Zhu ,&nbsp;Xueheng Zhao ,&nbsp;Lin Du ,&nbsp;Qianyin Zhou ,&nbsp;Lvjun Liu ,&nbsp;Shanshan Lv ,&nbsp;Huan Zhang ,&nbsp;Liqing Fan","doi":"10.1016/j.freeradbiomed.2025.08.061","DOIUrl":"10.1016/j.freeradbiomed.2025.08.061","url":null,"abstract":"<div><div>Human spermatogenesis is an important physiological process related to programmed cell death. However, which type of programmed cell death playing a key role in normal and abnormal human spermatogenesis remains obscure. This study integrated single-cell, bulk RNA and spatial transcriptome data analysis and found that the ferroptosis signal plays a potential role in spermatogenesis and significantly elevate in testicular samples from humans with non-obstructive azoospermia (NOA) due to various factors. In addition, the increased levels of GPX4-dependent ferroptosis in Sertoli cells is a common feature of NOA samples and inhibiting ferroptosis of Sertoli cells in azoospermia patients can improve the functional status of Sertoli cells. Based on weighted correlation network analysis (WGCNA), correlation analysis, differential expression analysis and <em>in vitro</em> experiments, we confirmed that long non-coding RNA (lncRNA) LINC00467 is a key factor in regulating ferroptosis in Sertoli cells. Finally, quantitative real-time polymerase chain reaction (qRT-PCR) experiments showed that LINC00467 can be used to predict sperm retrieval outcomes in NOA patients. In summary, this study reveals and verifies that the level of ferroptosis apparently elevates in Sertoli cells is a novel mechanism for NOA patients, and proposes that ferroptosis-related gene LINC00467 is potential biomarker and therapeutic target for NOA.</div></div>","PeriodicalId":12407,"journal":{"name":"Free Radical Biology and Medicine","volume":"240 ","pages":"Pages 650-662"},"PeriodicalIF":8.2,"publicationDate":"2025-08-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144948656","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}

{"title":"Depalmitoylation of TEAD1 facilitates lipid droplet accumulation and resistance to oxidative stress by transactivating PP2Acα","authors":"Xiaoli Sun ,&nbsp;Shuang Xu ,&nbsp;Yajie Ni ,&nbsp;Ruofan Chen ,&nbsp;Chunsun Dai","doi":"10.1016/j.freeradbiomed.2025.08.056","DOIUrl":"10.1016/j.freeradbiomed.2025.08.056","url":null,"abstract":"<div><h3>Background</h3><div>An overdose of acetaminophen (APAP) triggers acute liver failure via excessive production of reactive oxygen species (ROS). Modulating lipid droplet (LD) homeostasis in hepatocytes can protect against hepatic oxidative stress. However, rapid accumulation of LDs in the liver shortly after APAP administration remains unclear.</div></div><div><h3>Methods</h3><div>KEGG analysis was conducted to investigate the pathways associated with APAP-induced acute liver failure using data from the GSE database. Lipid metabolism-related pathways and the Hippo signaling pathway were identified as the most significantly enriched pathways. To investigate the functional role of Hippo signal in hepatotoxicity, hepatocyte-specific TEAD1 knockout mice were generated and challenged with APAP.</div></div><div><h3>Results</h3><div>Compared to wild-type controls, TEAD1-KO mice demonstrated significantly exacerbated hepatotoxicity, accompanied by reduced hepatic triglyceride (TG) content. Conversely, the hepatic overexpression of TEAD1 elevated TG levels and ameliorated APAP-induced liver injury. ChIP assays demonstrated that TEAD1 binds directly to the promoter region of PP2Acα, transcriptionally regulating its expression and promoting ACC1 dephosphorylation, thereby enhancing de novo lipogenesis. Furthermore, depalmitoylation of TEAD1 increased its capacity to form transcriptional condensates at the PP2Acα locus, resulting in enhanced PP2Acα transcriptional activity. This molecular mechanism facilitated the formation of numerous enlarged LDs, which conferred hepatoprotective effects against APAP toxicity.</div></div><div><h3>Conclusion</h3><div>Decreased palmitoylation of TEAD1 during the early stages of APAP administration facilitates rapid de novo lipid synthesis in response to lipid peroxidation. This process occurs through the enhanced capacity of TEAD1 for liquid-liquid phase separation (LLPS), which subsequently initiates transcription of the PP2Acα gene.</div></div>","PeriodicalId":12407,"journal":{"name":"Free Radical Biology and Medicine","volume":"240 ","pages":"Pages 532-545"},"PeriodicalIF":8.2,"publicationDate":"2025-08-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144925325","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}

{"title":"Cardiorespiratory fitness as a key predictor of metabolic, inflammatory, and oxidative stress biomarkers in adults with different physical activity levels","authors":"Omar A. Hernández-López ,&nbsp;Blanca Murillo-Ortíz ,&nbsp;Clara Luna-Marco ,&nbsp;Julia Cacace ,&nbsp;Alberto Hermo-Argibay ,&nbsp;Victoria Ramírez ,&nbsp;Martha Guevara-Cruz ,&nbsp;María Pelechá-Salvador ,&nbsp;Milagros Rocha ,&nbsp;Susana Rovira-Llopis ,&nbsp;Víctor M. Víctor ,&nbsp;Iván Torre-Villalvazo","doi":"10.1016/j.freeradbiomed.2025.08.058","DOIUrl":"10.1016/j.freeradbiomed.2025.08.058","url":null,"abstract":"<div><div>Sedentary lifestyles are associated with poor cardiorespiratory fitness (CRF) and predispose individuals to cardiometabolic diseases and increased all-cause mortality, events related to oxidative stress and inflammation. While regular exercise induces adaptations that improve metabolic homeostasis, its antioxidant effects are not fully characterised. This cross-sectional study compared antioxidant gene/protein expression in peripheral blood mononuclear cells (PBMCs) and metabolic, inflammatory, and oxidative stress biomarkers in sedentary and active individuals, and analysed potential associations with CRF.</div><div>Fifty-one healthy adults (18–45 years) were recruited. Participants were classified as sedentary (SED), physically active (PA), or endurance athletes (EA) using the International Physical Activity Questionnaire (IPAQ; n = 17/group). Anthropometric, biochemical, metabolic, and inflammatory variables were assessed. Oxidative stress was measured via serum hydrogen peroxide (H₂O₂) and malondialdehyde (MDA). PBMC protein/mRNA expression of nuclear factor erythroid 2-related factor 2 (NRF2) and antioxidant enzymes [superoxide dismutase 1 (SOD1), glutathione reductase (GSR), glutathione peroxidase 1 (GPX1), catalase (CAT)] were evaluated. Metabolic flexibility was assessed at rest by indirect calorimetry, and CRF by maximal oxygen uptake (VO₂max mL·kg⁻¹·min⁻¹) during a progressive maximal cardiopulmonary exercise test.</div><div>The PA and EA groups showed greater metabolic flexibility and CRF than the SED group, which exhibited altered homeostasis model assessment of insulin resistance (HOMA-IR), triglyceride-to-high-density lipoprotein cholesterol ratio (TG/HDL-C), elevated tumour necrosis factor alpha (TNFα), high-sensitivity C-reactive protein (hsCRP), oxidative stress (H₂O₂, MDA), and reduced antioxidant gene/protein expression. Higher VO₂max mL·kg⁻¹·min⁻¹ correlated with healthier metabolic profiles, less inflammation, and higher antioxidant expression, while inversely correlating with HOMA-IR, MDA, and TNFα.</div><div>Optimal or high CRF strongly protects against insulin resistance, oxidative stress, inflammation, and metabolic inflexibility, key hallmarks of sedentary behaviour. Regular physical exercise improves metabolic and redox profiles, enhancing antioxidant defences in PBMCs. A functional CRF threshold represents a practical target by which to reduce cardiometabolic risk.</div></div>","PeriodicalId":12407,"journal":{"name":"Free Radical Biology and Medicine","volume":"240 ","pages":"Pages 735-744"},"PeriodicalIF":8.2,"publicationDate":"2025-08-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144948592","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}

{"title":"Redox regulator LanCL1 suppresses glioma progression by coordinately inhibiting cell growth and regulating mitochondrial metabolism","authors":"Yunbo Yuan ,&nbsp;Junhong Li ,&nbsp;Mengping Wang ,&nbsp;Qiuyun Yuan ,&nbsp;Yanhui Liu ,&nbsp;Wanchun Yang ,&nbsp;Mina Chen","doi":"10.1016/j.freeradbiomed.2025.08.059","DOIUrl":"10.1016/j.freeradbiomed.2025.08.059","url":null,"abstract":"<div><div>Gliomas are highly aggressive and heterogeneous brain tumors with poor clinical outcomes, necessitating an urgent need for novel prognostic biomarkers and therapeutic targets. Redox regulation, which balances reactive oxygen species (ROS) generation with antioxidant defense mechanisms, has emerged as a crucial adaptive mechanism supporting glioma progression. However, the precise roles and clinical implications of redox-associated genes in glioma remain poorly defined. Here, we employed integrative analyses to explore the functional impact of redox-related genes in glioma and identified LanCL1 as a key regulator in glioma malignancy. Consensus clustering of 97 redox-related genes stratified gliomas into prognostic subtypes, with high LanCL1 expression correlating with low tumor grade, favorable molecular features (e.g., IDH1 mutation, 1p/19q co-deletion), and superior patient survival. Functional assays further revealed that LanCL1 inhibited glioma cell growth and migration by coordinately activating mitochondrial metabolism at the transcriptional level and inducing cell cycle arrest via modulation of CDK1/p27 axis. These findings highlight the critical role of redox regulation in glioma pathogenesis and establish LanCL1 as both a prognostic biomarker and potential therapeutic target for exploiting redox vulnerabilities in glioma.</div></div>","PeriodicalId":12407,"journal":{"name":"Free Radical Biology and Medicine","volume":"240 ","pages":"Pages 559-565"},"PeriodicalIF":8.2,"publicationDate":"2025-08-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144948649","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}

{"title":"Targeting KEAP1/NRF2 interaction with oleuropein ameliorates atherosclerosis by inhibiting macrophage ferroptosis","authors":"Xinxin Zhu ,&nbsp;Yuwu Chen ,&nbsp;Biyi Xu ,&nbsp;Junke Mou ,&nbsp;Mengyang Wang ,&nbsp;Qishuo Gu ,&nbsp;Qianhui Sun ,&nbsp;Man Li ,&nbsp;Chen Zhao ,&nbsp;Ming Zeng ,&nbsp;Ying Lv ,&nbsp;Shan Zhang ,&nbsp;Xiaoxuan Bai ,&nbsp;Jie Du ,&nbsp;Hang Yu ,&nbsp;Minghao Liu ,&nbsp;Xing Luo ,&nbsp;Ji Li ,&nbsp;Sining Hu ,&nbsp;Haibo Jia ,&nbsp;Bo Yu","doi":"10.1016/j.freeradbiomed.2025.08.036","DOIUrl":"10.1016/j.freeradbiomed.2025.08.036","url":null,"abstract":"<div><div>Atherosclerosis (AS) is a chronic inflammatory arterial disease. Oleuropein (OL), extracted from olive leaves, has demonstrated broad cardioprotective properties. However, the effects of OL on AS remain to be fully elucidated. ox-LDL incubated macrophages were used to imitate macrophage damage within plaques <strong><em>in vitro</em></strong>. Recombinant adeno-associated virus serotype 9 (AAV9) encoding a short hairpin RNA targeting NRF2 (AAV9-shNRF2) and AAV-KEAP1-R451S were administered to OL-treated ApoE^−/− mice. The molecular mechanisms were explored through immunoprecipitation and Chip-qPCR, seahorse assays, and proteomics analysis. OL administration significantly attenuated the progression of AS and enhanced plaque stability, as evidenced by reduced plaque area and lipid deposition, along with increased collagen content. Proteomics revealed that OL suppressed ferroptosis by upregulating GPX4/xCT level, improving mitochondrial function, alleviating oxidative stress and suppressing lipid peroxidation in macrophages treated by ox-LDL and atherosclerotic plaques. Moreover, OL enhanced NRF2 activation and nuclear translocation, while NRF2 inhibition or knockdown abolished the protective effect of OL on AS and macrophage ferroptosis. Mechanistically, molecular dynamics analysis suggested that OL may bind to the Arg415 site of KEAP1 competitively, promoting the separation and nuclear translocation of NRF2 from KEAP1. Additionally, transfection with the Arg415 mutant plasmid (KEAP1-R415S) abolished the antioxidative and anti-ferroptosis effects of OL in macrophages induced by ox-LDL. Moreover, the protective effects of OL against atherosclerosis and macrophage ferroptosis were significantly attenuated in AAV-KEAP1-R415S mutant ApoE^−/− mice. OL attenuated AS progression and macrophage ferroptosis by facilitating NRF2 nuclear translocation and activation via binding to the Arg415 residue of KEAP1 competitively. These findings identified a novel insight into potential therapeutic strategies for the treatment of AS.</div></div>","PeriodicalId":12407,"journal":{"name":"Free Radical Biology and Medicine","volume":"240 ","pages":"Pages 566-582"},"PeriodicalIF":8.2,"publicationDate":"2025-08-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144948622","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}

{"title":"NFATc1-targeted siRNA microdroplets prevent radiation-induced osteoporosis by coordinating bone-immune crosstalk","authors":"Yi Zhang ,&nbsp;Lijia Zhang ,&nbsp;Xilai Yang ,&nbsp;Xiliang Liu ,&nbsp;Ruimin Liu ,&nbsp;Wenqing Huang ,&nbsp;Yi Wang ,&nbsp;Yaping Ma ,&nbsp;Yin Xiao ,&nbsp;Jincheng Sima ,&nbsp;Xin Wang","doi":"10.1016/j.freeradbiomed.2025.08.057","DOIUrl":"10.1016/j.freeradbiomed.2025.08.057","url":null,"abstract":"<div><div>Radiation-induced bone loss, driven by osteoclast activation, involves the transcription factor nuclear factor of activated T-cells cytoplasmic 1 (NFATc1)-mediated signaling. This study developed NFATc1 siRNA-loaded microdroplets (NFATc1/MDs) to mitigate skeletal damage post-radiotherapy.</div></div><div><h3>Methods</h3><div>NFATc1/MDs were synthesized and characterized using TEM and confocal microscopy. Biocompatibility was tested in hBMSCs and RAW 264.7 macrophages. Osteoclastogenesis, osteogenesis, and adipogenesis were assessed <em>in vitro</em>, and therapeutic efficacy was evaluated in a rat radiation-induced bone loss model.</div></div><div><h3>Results</h3><div>NFATc1/MDs exhibited a core-shell structure, high biocompatibility, and efficient cellular uptake. They suppressed osteoclastogenesis without impairing osteogenic/adipogenic differentiation and modulated macrophage activity post-irradiation. <em>In vivo</em>, NFATc1/MDs preserved bone microstructure, reduced osteoclast numbers, and downregulated NFATc1, cathepsin K (CTSK), and tumor necrosis factor-alpha (TNF-α) expression. <strong>Conclusions</strong>: NFATc1/MDs effectively inhibited osteoclast-mediated bone resorption and inflammation, offering a targeted strategy to prevent radiation-induced bone loss. This biocompatible platform demonstrates potential for clinical translation in radiotherapy-associated skeletal complications.</div></div>","PeriodicalId":12407,"journal":{"name":"Free Radical Biology and Medicine","volume":"240 ","pages":"Pages 546-558"},"PeriodicalIF":8.2,"publicationDate":"2025-08-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144948675","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}

{"title":"Inhalation of Electrophilic and Redox Active Electronic Cigarette Aerosol Increases Oxidative Potential in the Lung in an Acute Manner.","authors":"David H Gonzalez, Ella M Kang, Emily N Nguyen, Jocelyn A Castellanos, Allen Louie, Rajat Gupta, Haoxuan Chen, Yifang Zhu, Steven J Campbell, Suzanne E Paulson, Maria C Jordan, Kenneth P Roos, Julian Whitelegge, Arthur K Cho, Jesus A Araujo","doi":"10.1016/j.freeradbiomed.2025.08.048","DOIUrl":"https://doi.org/10.1016/j.freeradbiomed.2025.08.048","url":null,"abstract":"<p><p>Electronic cigarettes (EC) are linked to pulmonary health effects involving oxidative stress, but underlying mechanisms are poorly understood. Induction of oxidative stress is thought to be mediated by imbalanced reactive oxygen species (ROS) production is implicated in the onset of inflammation in a multitude of pathogenic and diseases conditions. EC aerosol components deposited in the lungs can induce ROS generation and form electrophilic protein adducts, thereby promoting prooxidative redox environments in the airways. A variety of methods to assess oxidative stress exist but many are non-specific and have unclear molecular and/or biological interpretations confounding the understanding of mechanisms underlying EC toxicity. Here, we developed a method to detect prooxidant shifts in the redox environment of murine bronchoalveolar lavage fluid (BALF) following acute exposure to EC aerosol, using the terephthalate (TA) probe. Using acellular assays, we determined that the EC aerosol employed in this work has intrinsic electrophilic potential and capacity to generate •OH acellularly, suggesting that EC aerosols can promote prooxidative conditions by themselves. We then mathematically derived a parameter called the Iron (Fe)-Induced Oxidative Potential in BALF (FIBOP) that was calculated by measuring •OH generation rates in mixtures of 1-10 μM Fe(II) and 200 μM ascorbate in BALF. •OH generation rates normalized by total protein plotted as a function of [Fe(II)] allowed us to quantify FIBOP, 24 hours after acute exposure to EC. C57BL/6J mice exposed to EC aerosol for a total of 30 minutes in one day exhibited 60% enhancement of FIBOP compared to control mice exposed to filtered air (FA). This was accompanied by significantly increased levels of the free oxidized lipids, total HETEs (5-,12-,15-HETEs) (p<0.05) and total HODEs (9-,13-HODEs) (p<0.05) in BALF. Our results indicate that our method is sensitive to detecting prooxidant shifts in BALF induced by EC aerosol exposure in an acute manner.</p>","PeriodicalId":12407,"journal":{"name":"Free Radical Biology and Medicine","volume":" ","pages":""},"PeriodicalIF":8.2,"publicationDate":"2025-08-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144948590","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}

{"title":"MicroRNA-15b promotes myofascial pain syndrome by targeting NFS1 to regulate mitophagy","authors":"Dawei Han ,&nbsp;Molei Liu ,&nbsp;Mingwei Sheng ,&nbsp;Lili Jia ,&nbsp;Yunxia Liu ,&nbsp;Ling Liu ,&nbsp;Hongxia Li ,&nbsp;Yiqi Weng ,&nbsp;Xiaofei Song ,&nbsp;Yinghui Ren ,&nbsp;Wenli Yu","doi":"10.1016/j.freeradbiomed.2025.08.054","DOIUrl":"10.1016/j.freeradbiomed.2025.08.054","url":null,"abstract":"<div><h3>Background and aims</h3><div>Myofascial pain syndrome (MPS), driven by dysfunction in myofascial trigger points (MTrPs), remains mechanistically unclear. This study aimed to explore miR-15 b's function in MTrP pathogenesis, focusing on its regulation of iron-sulfur (Fe–S) cluster synthesis and mitophagy.</div></div><div><h3>Methods</h3><div>A rat MTrP model was established using repetitive mechanical injury and eccentric exercise. Skeletal muscle tissues and primary satellite cells were analysed for miR-15b expression, Fe-S cluster-related proteins (NFS1, NDUFS3, and SDH B), and mitophagy markers (FUN14 structural domain-containing protein 1 (FUNDC1) and LC3-II/I). <em>In vitro</em>, tumour necrosis factor-alpha (TNF-α)-induced inflammation and miR-15b modulation (mimics/sponges) and NFS1 modulation (overexpression/knockdown) were used to assess mitochondrial functions. <em>In vivo</em>, the therapeutic effect on normal and MTrP model rats was evaluated by intramuscular injection of transiently transfected complexes of NFS1 plasmid, miR-15b plasmid or sponge constructs.</div></div><div><h3>Results</h3><div>MTrP rats exhibited miR-15b, suppressed NFS1, and impaired Fe-S-dependent complexes. Dual luciferase assays verified miR-15b targeting NFS1. Rescue experiments further validated that miR-15b directly inhibits NFS1, increase reactive oxygen species (ROS), lowering mitochondrial membrane potential (MMP), triggering FUNDC1-mediated mitophagy. TNF-α stimulation elevated miR-15b levels, exacerbating mitochondrial dysfunction, whereas miR-15b inhibition restored NFS1 and normalised mitophagy. In normal rats, miR-15b overexpression recapitulated MTrP-like pathology in healthy rats. Moreover, in the MTrP model rats, miR-15b overexpression exacerbated these manifestations, sponge and NFS1 treatment attenuated or even reversed certain pathological changes.</div></div><div><h3>Conclusions</h3><div>miR-15b drives MTrP progression by suppressing NFS1, disrupting Fe-S homeostasis, and activating FUNDC1-dependent mitophagy. Targeting miR-15b mitigates mitochondrial dysfunction and pain hypersensitivity, underscoring its therapeutic potential in MPS.</div></div>","PeriodicalId":12407,"journal":{"name":"Free Radical Biology and Medicine","volume":"240 ","pages":"Pages 626-640"},"PeriodicalIF":8.2,"publicationDate":"2025-08-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144948680","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}