Free Radical Biology and Medicine最新文献

{"title":"Edaravone mitigates TDP-43 mislocalization in human amyotrophic lateral sclerosis neurons with potential implication of the SIRT1-XBP1 pathway","authors":"Satsuki Mikuriya,&nbsp;Tomo Takegawa-Araki,&nbsp;Makoto Tamura","doi":"10.1016/j.freeradbiomed.2025.01.012","DOIUrl":"10.1016/j.freeradbiomed.2025.01.012","url":null,"abstract":"<div><div>Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disorder characterized by progressive motor neuron loss along with pathological mislocalization of TAR DNA-binding protein 43 (TDP-43), a protein implicated in RNA metabolism. Although edaravone, a free-radical scavenger, has been approved for ALS treatment, its precise mechanism of action is not fully understood, particularly in relation to TDP-43 pathology. Here, we investigated the effects of edaravone on induced pluripotent stem cell (iPSC)-derived motor neurons in a patient with ALS harboring a TDP-43 mutation. Our results demonstrated that edaravone significantly attenuated neurodegeneration, as evidenced by neurite preservation, neuronal cell death reduction, and correction of aberrant cytoplasmic localization of TDP-43. These neuroprotective effects were not observed with vitamin C, indicating a unique mechanism of action for edaravone, distinct from its antioxidative properties. RNA sequencing revealed that edaravone rapidly modulated gene expression, including protein quality control pathway, such as the ubiquitin-proteasome system. Further analysis identified X-box binding protein (XBP1), a key regulator of the endoplasmic reticulum stress response, as a critical factor in the therapeutic effects of edaravone. This study suggests that edaravone may offer a multifaceted therapeutic approach for ALS by targeting oxidative stress and TDP-43 mislocalization through distinct molecular pathways.</div></div>","PeriodicalId":12407,"journal":{"name":"Free Radical Biology and Medicine","volume":"230 ","pages":"Pages 283-293"},"PeriodicalIF":7.1,"publicationDate":"2025-02-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143487633","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"HnRNPA2B1 promotes cardiac ferroptosis via m6A-dependent stabilization of PFN2 mRNA in myocardial ischemia-reperfusion injury.","authors":"Shuotao Shi, Qi Chen, Ying Yang, Zipei Li, Ruiyan Zheng, Rong Zhang, Zhongqiu Liu, Yuanyuan Cheng","doi":"10.1016/j.freeradbiomed.2025.02.038","DOIUrl":"https://doi.org/10.1016/j.freeradbiomed.2025.02.038","url":null,"abstract":"<p><p>Myocardial ischemia-reperfusion damage (MIRI) is a clinical problem and lacks proven treatment approaches. As a m6A reader, hnRNPA2B1 controls RNA destiny in the pathophysiology of neurodegenerative and cancerous disorders. Recently, we found that the level of hnRNPA2B1 was elevated in patients with myocardial infarction after percutaneous coronary intervention (PCI), which was positively correlated with cTnI. However, the role of hnRNPA2B1 in MIRI is still unknown. In the present study, we investigated the mechanism underlying MIRI-induced ferroptosis by focusing on a novel function of hnRNPA2B1. Our results showed that HnRNPA2B1 was also significantly increased in cardiomyocytes of MIRI models in vitro and in vivo. Genetically deleting hnRNPA2B1 effectively mitigated myocardial injury and cardiac function during MIRI. Silencing hnRNPA2B1 in cardiomyocytes boosted cell survival and decreased ferroptosis by lowering lipid ROS, MDA, Fe2+, and raising GSH, FTH1 levels, while overexpressing hnRNPA2B1 had the opposite impact. Mechanistic investigations revealed that hnRNPA2B1 recognized and interacted with the m6A site of PFN2 mRNA at \"AGACU\" to enhance the stability of PFN2 mRNA transcripts. Furthermore, PFN2 knockdown resulted in decreased MDA and Fe²⁺ levels and an increase in FTH1 expression. Importantly, silencing PFN2 attenuated ferroptosis in cardiomyocytes overexpressing hnRNPA2B1 during OGD/R injury. Collectively, hnRNPA2B1 potentially acts as a therapeutic target of MIRI through regulating caridac ferroptosis mediated by m6A-PFN2/FTH1 pathway.</p>","PeriodicalId":12407,"journal":{"name":"Free Radical Biology and Medicine","volume":" ","pages":""},"PeriodicalIF":7.1,"publicationDate":"2025-02-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143515108","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":"Maternal lactoferrin supplementation prevents mitochondrial and redox homeostasis dysfunction, and improves antioxidant defenses through Nrf2 and UCP2 signaling after neonatal hypoxia-ischemia","authors":"Andrey Vinicios S. Carvalho ,&nbsp;Eduardo F. Sanches ,&nbsp;Rafael T. Ribeiro ,&nbsp;Luz Elena Durán-Carabali ,&nbsp;Osmar Ramires Júnior ,&nbsp;Bruna Dutra Muniz ,&nbsp;Moacir Wajner ,&nbsp;Angela T. Wyse ,&nbsp;Carlos Alexandre Netto ,&nbsp;Stéphane V. Sizonenko","doi":"10.1016/j.freeradbiomed.2025.02.036","DOIUrl":"10.1016/j.freeradbiomed.2025.02.036","url":null,"abstract":"<div><div>Neonatal hypoxia-ischemia (HI) is a major cause of mortality and neurological impairments in infants. Main HI-induced pathological mechanisms include mitochondrial dysfunction and oxidative stress due to insufficient oxygen and energetic substrates to the nervous cells. Bovine lactoferrin (Lf) has demonstrated neuroprotective effects in several experimental models of neonatal brain injury in rodents, however its mechanisms remain unclear. This study aimed to evaluate the early impact of maternal dietary supplementation with Lf on redox and hippocampal mitochondrial function following neonatal HI. From postnatal day 6 (PND6), pregnant Wistar rats were fed with a diet supplemented with Lf (1 g/kg) or with an isocaloric control diet until offspring euthanasia. At PND7, pups of both sexes were subjected to experimental HI through the occlusion of the right common carotid artery followed by 60 min of hypoxia (8 % oxygen). Lf prevented HI-induced increased levels of DCFH and lipoperoxidation in hippocampus. Furthermore, Lf enhanced antioxidant defenses including SOD, GPx, and GSH, counteracting HI-induced oxidative stress. HI injury altered the activities of enzymes in the mitochondrial respiratory chain and increased the mitochondrial membrane potential. Both effects were counteracted by Lf supplementation. Lactoferrin prevented oxidative stress and to restored mitochondrial function by upregulating Nrf2 and UCP2 expression following experimental HI. Our results show that even a shorter period of Lf delivery to rat pups is able to improve hippocampal response to neonatal hypoxia-ischemia, reversing initial mechanisms of damage in the cascade of HI injury.</div></div>","PeriodicalId":12407,"journal":{"name":"Free Radical Biology and Medicine","volume":"231 ","pages":"Pages 68-79"},"PeriodicalIF":7.1,"publicationDate":"2025-02-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143509733","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":"Nuclear Factor κB Signaling Revisited: Its Role in Skeletal Muscle and Exercise.","authors":"Li Li Ji","doi":"10.1016/j.freeradbiomed.2025.02.013","DOIUrl":"https://doi.org/10.1016/j.freeradbiomed.2025.02.013","url":null,"abstract":"<p><p>Nuclear factor (NF) κB as a redox sensitive, anti-apoptotic and pro-inflammatory signaling molecule has been studied extensively for more than three decades. Its role in inducing antioxidant enzymes, defending against extracellular and intracellular stress and maintaining redox homeostasis in skeletal muscle has also been recognized. New research continues to explore the polytropic nature of NFκB in cellular function, especially its crosstalk with other important signaling pathways. Understanding of the broad impact of these functions has significant implications in health and disease of skeletal muscle as an organ designed for contraction and mobility. Two important aspects of muscle wellbeing, i.e., disease and aging, are not discussed in this review. This review will provide an update on the new findings related to NFκB involvement in multiple signaling pathways and refresh our knowledge of its activation in skeletal muscle with a special reference to physical exercise.</p>","PeriodicalId":12407,"journal":{"name":"Free Radical Biology and Medicine","volume":" ","pages":""},"PeriodicalIF":7.1,"publicationDate":"2025-02-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143515110","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":"Identifying genetic overlaps in obesity and metabolic disorders unlocking unique and shared mechanistic insights","authors":"Liwan Fu ,&nbsp;Xiaodi Han ,&nbsp;Yuquan Wang ,&nbsp;Yue-Qing Hu","doi":"10.1016/j.freeradbiomed.2025.02.033","DOIUrl":"10.1016/j.freeradbiomed.2025.02.033","url":null,"abstract":"<div><h3>Objective</h3><div>Obesity has a high heritability and frequently co-occurs with metabolic disorders, indicating shared genetic susceptibility. The underlying causative genes and biological mechanisms of obesity and metabolic disorders remain predominantly elusive.</div></div><div><h3>Methods</h3><div>The FinnGen R11 dataset, including over 450,000 subjects, was employed in conjunction with the Genotype-Tissue Expression Project (GTEx) v8 eQTls dataset to conduct cross-tissue transcriptome association studies, Functional Summary-based Imputation in single tissues, and Gene Analysis combined with Multimarker Analysis of Genomic Annotation, respectively, for identifying distinct and shared genetic architectures of obesity and metabolic disorders. We also employed RHOGE to ascertain the genetic correlation and putative causal directions between them. Subsequent Mendelian randomization, colocalization analyses, and other cell and tissue enrichment analyses were employed to enhance our understanding of the functional implications of these susceptibility genes.</div></div><div><h3>Results</h3><div>A total of 35 genes were identified as obesity susceptibility and 10 genes linked to metabolic disorder susceptibility. Of these, three genes (<em>MCM6</em>, <em>MAPRE3</em> and <em>UBXN4</em>) were identified as being shared. Mendelian randomization and colocalization analyses revealed the three shared genes have causal associations with obesity and metabolic disorders and serve as independent signals. Subsequent analyses indicated <em>MCM6</em> may influence obesity and metabolic disorder risk by regulating DNA replication, cell proliferation, and interactions with chemical responses. <em>MAPRE3</em> may confer protective effects against obesity and metabolic disorders through PAK Pathway, while <em>UBXN4</em> may involve in regulating cholesterol metabolism.</div></div><div><h3>Conclusion</h3><div>Our study provides insight into the novel shared genetic mechanism between obesity and metabolic disorders and identifies potential targets for pharmacological intervention.</div></div>","PeriodicalId":12407,"journal":{"name":"Free Radical Biology and Medicine","volume":"231 ","pages":"Pages 80-93"},"PeriodicalIF":7.1,"publicationDate":"2025-02-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143500366","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":"Chlorogenic acid improves SPS-induced PTSD-like behaviors in rats by regulating the crosstalk between Nrf2 and NF-κB signaling pathway.","authors":"Can Tang, Jie Gao, Sen Li, Hui Cheng, Yu-Yuan Peng, Yang Ding, Huan Yang, Xin-Mei Ma, Hai-Yan Wang, Zai-Yun Long, Xiu-Min Lu, Yong-Tang Wang","doi":"10.1016/j.freeradbiomed.2025.02.034","DOIUrl":"https://doi.org/10.1016/j.freeradbiomed.2025.02.034","url":null,"abstract":"<p><p>Post-traumatic stress disorder (PTSD) is a long-term delayed mental disorder caused by sudden, threatening or catastrophic life events. Chlorogenic acid (CGA) is a polyphenolic acid rich in Eucommia ulmoides and other plants with potential neuroprotective effects, effectively enhances learning and memory, and exerts a beneficial impact on improving mood and attention. However, the effects and mechanisms of CGA on PTSD-like behaviors remain uncertain. This study is to explore the effects and mechanisms of CGA on PTSD by using network pharmacology analysis, molecular docking and experimental validation, and try to provide new strategies for the treatment of PTSD. The results indicated that 9 core targets with a strong binding affinity with CGA were screened out, and they were mainly enriched in apoptosis, inflammation, and oxidative stress. The followed vivo experiments indicated that CGA could alleviate single prolonged stress (SPS)-induced PTSD-like behaviors, and improve hippocampal pathological damage, apoptosis and synaptic plasticity through antioxidant and anti-inflammatory effects by regulating Nrf2 and NF-κB pathways. Thus, CGA may inhibit hippocampal neuronal apoptosis, reduce neuroinflammatory and oxdiative stress response, and enhance hippocampal synaptic plasticity through regulating the crosstalk between Nrf2 and NF-κB signaling pathway, thereby improving SPS-induced PTSD-like behaviors.</p>","PeriodicalId":12407,"journal":{"name":"Free Radical Biology and Medicine","volume":" ","pages":""},"PeriodicalIF":7.1,"publicationDate":"2025-02-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143500363","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":"A superoxide anion responsive and self-reporting fluorescent H2S donor for the treatment of diabetic wound","authors":"Ning Zhang ,&nbsp;Tengchuan Fu ,&nbsp;Tao Li ,&nbsp;Pengjie Zhong ,&nbsp;Luoyi Li ,&nbsp;Mingtao Peng ,&nbsp;Zhenghao Li ,&nbsp;Ling Zhang ,&nbsp;Han Wang ,&nbsp;Ping Hu ,&nbsp;Yifei Lu ,&nbsp;Mengyun Yao","doi":"10.1016/j.freeradbiomed.2025.02.026","DOIUrl":"10.1016/j.freeradbiomed.2025.02.026","url":null,"abstract":"<div><div>Superoxide anion (O₂^•−) not only serves as a critical precursor for numerous damaging reactive oxygen species (ROS), but also is implicated in a variety of diseases, including cancer, cardiovascular disorders, and diabetes. Consequently, reducing the levels of superoxide anions and alleviating oxidative stress are of paramount importance. Conversely, hydrogen sulfide (H₂S), recognized as a significant biological signaling molecule, plays vital roles in protecting mammalian cells from oxidative damage and promoting tissue regeneration. In this study, we reported a novel superoxide anion-responsive H₂S donor (HSD-SO-B) designed to scavenge O₂^•− and produce H₂S concurrently. This H₂S donor exhibits several advantages: (1) rapid response to superoxide anions (O₂^•−) with remarkable selectivity over competing species (2) generating H₂S while scavenging superoxide anions (3) producing ratiometric fluorescence for both visualization and quantification of H₂S release. Moreover, this O₂^•−-responsive, self-immolative fluorescent H₂S donor has shown significant therapeutic and reparative effects on the diabetic wound model in mice.</div></div>","PeriodicalId":12407,"journal":{"name":"Free Radical Biology and Medicine","volume":"231 ","pages":"Pages 109-119"},"PeriodicalIF":7.1,"publicationDate":"2025-02-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143476423","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":"HepG2 cells do not express xanthine oxidoreductase (XOR): Implications for XOR and uric acid-related research.","authors":"King R, Chapa M, Chen I, Giromini A, E E Kelley, Khoo N K H","doi":"10.1016/j.freeradbiomed.2025.02.023","DOIUrl":"https://doi.org/10.1016/j.freeradbiomed.2025.02.023","url":null,"abstract":"<p><p>Although the relative extent of xanthine oxidoreductase (XOR) varies considerably in human tissues, the greatest specific activity is reported in the liver and intestines. Unlike murine models, where primary hepatocytes are readily available, human counterparts are not. As such, investigators often utilize the human carcinoma cell line HepG2 for in vitro experimentation as these cells proliferate well in culture medium. Some of the studies using HepG2 cells for proof-of-principal experimentation focus on uric acid (UA) and/or XOR activity. However, it has been reported that hepatocellular carcinoma diminishes XOR expression to nearly unmeasurable levels when compared to normal cell counterparts which posits the question of validity in the context of using HepG2 cells for XOR/UA assessments. As such, we closely examined XOR expression, protein abundance, and enzymatic activity in HepG2 cells and compared these results to an immortalized murine hepatocyte line (AML12) as well as murine liver homogenate. We report the absence of detectable XOR message, protein, and enzymatic activity in HepG2 cells. Since cellular XOR expression has been reported to be stimulated by hypoxia and serum starvation, we then exposed both AML12 and HepG2 cells to hypoxia (1% O₂) for 24 h or serum starvation which impacted XOR activity in AML12 cells, but not in HepG2 cells. Knowing that several studies report XOR activity in HepG2 cells, we compared a commonly used non-selective XOR assay kit to our sensitive and selective HPLC/electrochemical assay. The kit assay indicated XOR activity in HepG2 cells; however, XOR inhibition did not diminish this activity. To account for alternative enzymatic oxidization of hypo/xanthine to UA, we investigated HepG2 cells for aldehyde oxidase (AO) protein abundance and found immunodetectable AO protein as well as AO-inhibitable conversion of hypoxanthine to UA; albeit, at a much slower rate when compared to XOR. In toto, these data confirm that HepG2 cells do not demonstrate XOR activity and thus commonly used kit assays for XOR, in the absence of XOR inhibitor controls, may indicate a positive response induced by an alternative source of peroxide and UA such as AO.</p>","PeriodicalId":12407,"journal":{"name":"Free Radical Biology and Medicine","volume":" ","pages":""},"PeriodicalIF":7.1,"publicationDate":"2025-02-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143482614","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":"Methylglyoxal deteriorates macrophage efferocytosis in diabetic wounds through ROS-induced ubiquitination degradation of KLF4","authors":"Hanting Zhu ,&nbsp;Wenao Wang ,&nbsp;Jiajun Zhu,&nbsp;Xuelian Chen,&nbsp;Jizhuang Wang,&nbsp;Jiaqiang Wang,&nbsp;Dan Liu,&nbsp;Peilang Yang,&nbsp;Yan Liu","doi":"10.1016/j.freeradbiomed.2025.02.030","DOIUrl":"10.1016/j.freeradbiomed.2025.02.030","url":null,"abstract":"<div><div>Diabetic wounds are a leading cause of disability and mortality in patients with diabetes, and persistent low-grade inflammation plays a significant role in their pathogenesis. Methylglyoxal (MGO), an active product of glucose metabolism, often induces chronic inflammation and is considered a major risk factor in the healing of diabetic wounds. Efferocytosis, the process by which macrophages clear apoptotic cells, is crucial for terminating the inflammatory response and tissue repair. However, the role of MGO in macrophage efferocytosis remains unclear. This study aimed to investigate whether MGO regulates macrophage efferocytosis and the underlying mechanisms. In this study, we observed impaired efferocytosis in diabetic wounds, leading to the accumulation of apoptotic neutrophils and a relative deficiency of M2 macrophages, with MGO being a significant cause. MGO promotes the production of ROS, which not only activates the MAPK p38 pathway, but also upregulates the transcription of the E3 ubiquitin ligase FBXO32, catalyzing the ubiquitination of the transcription factor KLF4 and suppressing the transcription of <em>MerTK</em> mRNA, thereby affecting the phagocytic function of macrophages. Inhibition of the MAPK p38 pathway or knockdown of FBXO32 reduced the ubiquitination and degradation of KLF4, thus mitigating the impairment of efferocytosis caused by oxidative stress. This study reveals the mechanism by which MGO inhibits efferocytosis in diabetic wounds, providing a new target and theoretical basis for the treatment of chronic diabetic wounds.</div></div>","PeriodicalId":12407,"journal":{"name":"Free Radical Biology and Medicine","volume":"231 ","pages":"Pages 23-37"},"PeriodicalIF":7.1,"publicationDate":"2025-02-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143471409","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Artemether relieves liver fibrosis by triggering ferroptosis in hepatic stellate cells via DHHC12-mediated S-palmitoylation of the BECN1 protein","authors":"Mengran Li ,&nbsp;Yuqi Sun ,&nbsp;Yuyao Wei ,&nbsp;Yujia Li ,&nbsp;Jiang Juan Shao ,&nbsp;Mei Guo ,&nbsp;Shizhong Zheng ,&nbsp;Zili Zhang","doi":"10.1016/j.freeradbiomed.2025.02.031","DOIUrl":"10.1016/j.freeradbiomed.2025.02.031","url":null,"abstract":"<div><div>Liver fibrosis, a pivotal stage in chronic liver disease progression, is driven by hepatic stellate cell (HSC) activation. Ferroptosis is a novel form of programmed cell death, which offers therapeutic potential for liver fibrosis. Although artemether (ART) exhibits antifibrotic properties, its mechanisms in liver fibrosis remain unclear. This study aimed to determine the therapeutic effects of ART on liver fibrosis and explore the role of S-palmitoylation in HSC ferroptosis.</div></div><div><h3>Methods</h3><div>A mouse model of liver fibrosis was constructed by carbon tetrachloride (CCl₄) injection. Transforming growth factor-β (TGF-β) was used for stimulating HSC activation in vitro. Histopathological and serological assays were performed to analyze the therapy effects of ART. Liquid Chromatography/Mass Spectrometry (LC/MS) and acyl-biotinyl exchange (ABE) were used to determine the role of S-palmitoylation in ART-induced HSC ferroptosis. Western blot and Co-Immunoprecipitation (Co-IP) were performed to examine the effects of autophagy in ART-induced HSC ferroptosis through regulating BECN1 S-palmitoylation.</div></div><div><h3>Results</h3><div>ART ameliorated liver fibrosis by inducing HSC ferroptosis, and the ferroptosis inhibitor ferrostatin-1 (Fer-1) impaired the inhibitory effect of ART. Interestingly, the levels of S-palmitoylation were elevated by upregulating the palmitoyltransferase DHHC12 during ART-induced HSC ferroptosis. DHHC12 knockdown reduced S-palmitoylation levels and impaired ART-mediated HSC ferroptosis. RNA-seq analysis indicated that autophagy activation was essential for ART to induce HSC ferroptosis. 3-methyladenine (3-MA) suppressed autophagy and ART-induced HSC ferroptosis. Importantly, BECN1 S-palmitoylation by DHHC12 drove ART to activate autophagy. DHHC12 bound to the cysteine 21 residue of BECN1, thereby stabilizing the BECN1 protein and facilitating autophagy activation. Mutation of the cysteine 21 residue decreased BECN1 protein stability, autophagy activation and ferroptosis in ART-treated HSCs. In a mouse model of hepatic fibrosis, HSC-specific inhibition of BECN1 S-palmitoylation reversed ART-induced HSC ferroptosis and the improvement of fibrotic liver.</div></div><div><h3>Conclusions</h3><div>ART alleviates liver fibrosis by inducing HSC ferroptosis via DHHC12-mediated BECN1 protein S-palmitoylation.</div></div>","PeriodicalId":12407,"journal":{"name":"Free Radical Biology and Medicine","volume":"231 ","pages":"Pages 120-135"},"PeriodicalIF":7.1,"publicationDate":"2025-02-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143482610","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}