The FASEB Journal最新文献

{"title":"ATF7–PINK1 Axis Governs Mitophagy and Intestinal Inflammation in Ulcerative Colitis","authors":"Yidong Chen,&nbsp;Xiaopeng Zhang,&nbsp;Junrong Li,&nbsp;Fang Liu,&nbsp;Qi Yu,&nbsp;Jiamin Li,&nbsp;Liangru Zhu","doi":"10.1096/fj.202500813R","DOIUrl":"https://doi.org/10.1096/fj.202500813R","url":null,"abstract":"<p>Ulcerative colitis (UC), a chronic inflammatory bowel disease, is marked by sustained inflammation and excessive apoptosis of intestinal epithelial cells (IECs). Despite progress in understanding UC pathogenesis, the role of activating transcription factors (ATFs) in disease progression remains elusive. Here, we profile the expression of ATF family members (ATF1–ATF7) in the colonic mucosa of UC patients and identify ATF7 as a critical regulator of mitophagy through its control of PTEN-induced kinase 1 (PINK1). Expression levels of ATF1–ATF7 were quantified in colonic mucosal samples from UC patients (<i>n</i> = 219) and healthy controls (<i>n</i> = 105) via quantitative PCR. Using IEC-specific ATF7 knockout mouse models and human CCD 841 CoN colonic epithelial cells, we employed ChIP-seq, dual-luciferase assays, transmission electron microscopy, and immunofluorescence to elucidate their roles in mitophagy and disease progression. Clinical correlation between ATF7 expression and disease severity was assessed using the Mayo score. ATF7 expression was significantly reduced in UC patients and inversely correlated with disease severity. Mechanistically, ATF7 was identified as a direct transcriptional activator of PINK1, a key mitophagy regulator. Loss of ATF7 or PINK1 disrupted mitophagy, exacerbating mitochondrial dysfunction, IEC apoptosis, and colonic inflammation in vivo and in vitro. Our findings uncover a pivotal ATF7-PINK1 axis that governs mitophagy and limits UC progression. The inverse correlation between ATF7 expression and UC severity highlights its potential as a therapeutic target, offering new avenues for intervention in this debilitating disease.</p>","PeriodicalId":50455,"journal":{"name":"The FASEB Journal","volume":"39 13","pages":""},"PeriodicalIF":4.4,"publicationDate":"2025-06-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1096/fj.202500813R","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144514469","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":"Deciphering the Effect of Ginsenoside Rk3 on Gut Microbiota and Its Applications","authors":"Ammu P. Nair,&nbsp;Deepika Kaushik,&nbsp;Ashwani Kumar,&nbsp;Jyoti Singh,&nbsp;Emel Oz,&nbsp;Avinash Sharma,&nbsp;Ali Jebreen,&nbsp;Charalampos Proestos,&nbsp;Fatih Oz,&nbsp;Mukul Kumar","doi":"10.1096/fj.202501234RR","DOIUrl":"https://doi.org/10.1096/fj.202501234RR","url":null,"abstract":"<div>\u0000 \u0000 <p>Gut microbiota, a complex community of microorganisms residing in the gastrointestinal tract, has been implicated in the overall health and disease states of the host organism. Among various bioactive compounds, ginsenoside Rk3, a component of the medicinal plant <i>Panax ginseng</i>, has emerged as a potential modulator of gut microbial composition. This abstract aimed to highlight the impact of ginsenoside Rk3 on gut microbiota, uncovering its effects on microbial diversity, abundance, and functionality. By investigating the specific changes induced by ginsenoside Rk3 within the gut microbial ecosystem, the study illuminates its potential therapeutic roles, thereby expanding the understanding of host–microbiome interactions and their significance in health and disease.</p>\u0000 </div>","PeriodicalId":50455,"journal":{"name":"The FASEB Journal","volume":"39 13","pages":""},"PeriodicalIF":4.4,"publicationDate":"2025-06-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1096/fj.202501234RR","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144514511","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":"Akkermansia muciniphila-Derived Outer Membrane Vesicles as a Novel Therapeutic Approach for Mastitis: Insights From In Vitro and Vivo Studies","authors":"Boqi Zhang,&nbsp;Nan Wang,&nbsp;Guitian He,&nbsp;Tong Chen,&nbsp;Jinxin Zong,&nbsp;Caomeihui Shen,&nbsp;Yueying Wang,&nbsp;Chuanghang Li,&nbsp;Xuanqi Yin,&nbsp;Yang Meng,&nbsp;Fuqiang Chang,&nbsp;Sihui Wang,&nbsp;Chunjin Li,&nbsp;Xu Zhou","doi":"10.1096/fj.202500877RR","DOIUrl":"https://doi.org/10.1096/fj.202500877RR","url":null,"abstract":"<p>Mastitis is a major disease affecting the dairy industry. Although antibiotics are the most prevalent treatment for mastitis, their overuse presents significant risks to public health. Probiotic therapy has emerged as a promising alternative for controlling mastitis, but there is a lack of comprehensive studies on the efficacy and mechanisms of specific probiotics in treating this condition. Here, we investigate the potential of <i>Akkermansia muciniphila</i> (<i>A. muciniphila</i>) and its derived outer membrane vesicles (AOMVs) as therapeutic agents for mastitis. Significant differences in microbial communities were identified through an analysis of the microbiota composition in milk from healthy and mastitis cows. Spearman's correlation analysis revealed a significant negative correlation between the relative abundances of <i>Verrucomicrobia</i> and <i>Akkermansia</i> and the levels of inflammatory cytokines in mastitis milk. Furthermore, we evaluated the roles of live and pasteurized <i>A. muciniphila</i> in lipopolysaccharide (LPS)-induced inflammatory responses in bovine mammary epithelial cells (MAC-T). Compared to pasteurized <i>A. muciniphila</i>, live <i>A. muciniphila</i> exhibited a stronger anti-inflammatory effect, notably inhibiting the TLR4 and NF-κB signaling pathways. We also demonstrated that these effects of <i>A. muciniphila</i> may be mediated through AOMVs. Subsequently, the therapeutic potential of AOMVs was evaluated in vivo using an LPS-induced mastitis mouse model. The results demonstrated that AOMVs targeted the mammary glands, alleviated LPS-induced damage, and reduced inflammatory cytokine levels and signaling pathway activation. Finally, we conducted a preliminary investigation to assess the therapeutic effects of <i>A. muciniphila</i> on mastitis in dairy cows. Administration of <i>A. muciniphila</i> significantly reduced somatic cell count (SCC) in the milk of cows with mastitis. Notably, four of the five treated cows showed a decrease in SCC to below 2 × 10⁵ cells/mL, with negative results in the California Mastitis Test. Our findings provide compelling evidence for the therapeutic potential of <i>A. muciniphila</i> and AOMVs in the treatment of mastitis. These results not only highlight the anti-inflammatory properties of <i>A. muciniphila</i> as a potential probiotic, proposing novel strategies for developing alternatives to antibiotic therapies, but also provide a theoretical basis for exploring the role of outer membrane vesicles in the host-microbe interface.</p>","PeriodicalId":50455,"journal":{"name":"The FASEB Journal","volume":"39 13","pages":""},"PeriodicalIF":4.4,"publicationDate":"2025-06-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1096/fj.202500877RR","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144515050","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":"SRSF5 Regulates Trophoblast Apoptosis by Inhibiting NR2F2 Transcriptional Activity Through MLX Ubiquitin Degradation Mediated by Alternative Splicing in Preeclampsia","authors":"Ranhong Li,&nbsp;Rui Chen,&nbsp;Li Wang,&nbsp;Kan Liu,&nbsp;Haiying Wu","doi":"10.1096/fj.202501137R","DOIUrl":"https://doi.org/10.1096/fj.202501137R","url":null,"abstract":"<div>\u0000 \u0000 <p>The pathogenesis of preeclampsia (PE) is closely related to the dysfunction of placental trophoblast. However, the precise mechanisms underlying placental dysfunction remain inadequately elucidated. The expression of Serine/arginine-rich Splicing Factor 5 (SRSF5) in the placental tissues of PE mice was detected by quantitative real time-PCR (qRT-PCR), western blot and immunohistochemistry (IHC) experiments. The functions of SRSF5 in HTR8/SVneo cells were detected by CCK8, wound healing and transwell assays. TUNEL was used to analyze the apoptosis of HTR8/SVneo cells. Subsequently, alternative splicing events of Max-like protein X (MLX) were identified using RT-PCR and RIP assays. In addition, experiments such as Co-IP and in vivo ubiquitination were used to analyze the molecular mechanism by which SRSF5 regulates apoptosis of HTR8/SVneo cells. <b>S</b>RSF5 was significantly overexpressed in placental tissues of PE mice. Knockdown of SRSF5 induced proliferation, migration and invasion of HTR8/SVneo cells and inhibited cell apoptosis. Mechanistically, silencing SRSF5 could induce the ubiquitination and degradation of MLX through alternative splicing in HTR8/SVneo cells. Furthermore, the knockdown of SRSF5 enhanced the transcriptional activity of nuclear receptor subfamily 2 group F member 2 (NR2F2) via MLX, which contributed to the apoptosis of HTR8/SVneo cells. In summary, the downregulation of SRSF5 enhances the transcriptional activation of NR2F2 and inhibits trophoblast cell apoptosis by facilitating ubiquitination and degradation of MLX protein through alternative splicing, thereby participating in the pathogenesis of PE.</p>\u0000 </div>","PeriodicalId":50455,"journal":{"name":"The FASEB Journal","volume":"39 13","pages":""},"PeriodicalIF":4.4,"publicationDate":"2025-06-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144514512","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":"Effect of Renal Denervation on Early and Late Stages of Diabetic Nephropathy","authors":"Biyang Xu,&nbsp;Melissa Lowe,&nbsp;Vladislav Levchenko,&nbsp;Mark N. Morales,&nbsp;Christine A. Klemens,&nbsp;Christopher T. Banek,&nbsp;Alexander Staruschenko","doi":"10.1096/fj.202501453R","DOIUrl":"https://doi.org/10.1096/fj.202501453R","url":null,"abstract":"<p>Diabetic kidney disease (DKD) is a major driver of morbidity and mortality worldwide. Although the FDA has recently approved renal denervation (RDNx) to treat hypertension, contradictory findings have been reported on the effect of RDNx on the progression of DKD in type 2 diabetes. Here, we used type 2 diabetic nephropathy (T2DN) rats, a well-established non-obese model that mimics key features in human DKD, to evaluate the effect of RDNx on the progression of DKD in the early and late stages of diabetic nephropathy. Results indicate that RDNx lowers mean arterial pressure in the early but not late stage of DKD without improving glycemic control or renal function, which may suggest that the effect of RDNx on type 2 diabetes is stage-dependent, emphasizing the importance of timing when considering RDNx as a potential treatment for DKD.</p>","PeriodicalId":50455,"journal":{"name":"The FASEB Journal","volume":"39 13","pages":""},"PeriodicalIF":4.4,"publicationDate":"2025-06-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1096/fj.202501453R","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144503219","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":"Exploring PU.1 as a Therapeutic Target to Mitigate Oxidative Stress and Inflammation in Peritoneal Dialysis-Induced Peritoneal Fibrosis","authors":"Shuo Zhong,&nbsp;Wenyan Su,&nbsp;Yunzhao Wang,&nbsp;Dan Liu,&nbsp;Xiumei Li,&nbsp;Fangzhou Ma,&nbsp;Liang Xu,&nbsp;Jie Gao,&nbsp;Haiping Wang,&nbsp;Jing Sun","doi":"10.1096/fj.202500305RR","DOIUrl":"https://doi.org/10.1096/fj.202500305RR","url":null,"abstract":"<div>\u0000 \u0000 <p>The transcription factor PU.1 has been implicated in various fibrotic diseases; however, its role in peritoneal dialysis-associated peritoneal fibrosis (PF) remains unclear. The role of PU.1 was investigated using knockdown approaches with AAV9-shPU.1 in a rat PF model and siRNA in human peritoneal mesothelial cells (MeT-5A). Multiple experimental techniques, including real-time quantitative PCR, western blot, enzyme-linked immunosorbent assay (ELISA), and immunohistochemical staining, were utilized. In vivo, PU.1 expression was significantly upregulated in the rat PF model. Knockdown of PU.1 ameliorated peritoneal thickening, collagen deposition, angiogenesis, inflammation, and oxidative stress. In vitro, high-glucose exposure upregulated PU.1 expression in MeT-5A cells, promoting fibrosis and angiogenesis. PU.1 knockdown reduced fibrosis-related markers and suppressed the IL-6/JAK1/STAT3 signaling pathway, a critical regulator of PF. Treatment with an IL-6 inhibitor further confirmed that PU.1 modulates fibrosis through this pathway. PU.1 plays a pivotal role in the progression of PF. Its knockdown mitigates fibrosis by reducing inflammation and angiogenesis, and suppressing the activation of the IL-6/JAK1/STAT3 pathway, highlighting its potential as a therapeutic target.</p>\u0000 </div>","PeriodicalId":50455,"journal":{"name":"The FASEB Journal","volume":"39 13","pages":""},"PeriodicalIF":4.4,"publicationDate":"2025-06-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144503228","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":"Skeletal Muscle Mitochondria Contain Nuclear-Encoded RNA Species Prior to and Following Adaptation to Exercise Training in Rats","authors":"Jessica L. Silver,&nbsp;Séverine Lamon,&nbsp;Stella Loke,&nbsp;Gisella Mazzarino,&nbsp;Larry Croft,&nbsp;Mark Ziemann,&nbsp;Megan Soria,&nbsp;Glenn D. Wadley,&nbsp;Adam J. Trewin","doi":"10.1096/fj.202500157R","DOIUrl":"https://doi.org/10.1096/fj.202500157R","url":null,"abstract":"<p>Skeletal muscle mitochondria adaptation to exercise training is mediated by molecular factors that are not fully understood. Mitochondria import over 1000 proteins encoded by the nuclear genome, but the RNA population resident within the organelle is generally thought to be exclusively encoded by the mitochondrial genome. However, recent in vitro evidence suggests that specific nuclear-encoded miRNAs and other noncoding RNAs (ncRNAs) can reside within the mitochondrial matrix. Whether these are present in mitochondria of skeletal muscle tissue, and whether this is affected by endurance training—a potent metabolic stimulus for mitochondrial adaptation—remains unknown. Rats underwent 4 weeks of moderate-intensity treadmill exercise training, then were humanely killed and tissues were collected for molecular profiling. Mitochondria from gastrocnemius skeletal muscle were isolated by immunoprecipitation, further purified, and then the resident RNA was sequenced to assess the mitochondrial transcriptome. Exercise training elicited typical transcriptomic responses and functional adaptations in skeletal muscle, including increased mitochondrial respiratory capacity. We identified 24 nuclear-encoded coding or noncoding RNAs in purified mitochondria, in addition to 50 nuclear-encoded miRNAs that met a specified abundance threshold. Although none were differentially expressed in the exercise vs. control group at FDR &lt; 0.05, exploratory analyses suggested that the abundance of 3 miRNAs was altered (<i>p</i> &lt; 0.05) in mitochondria isolated from trained compared with sedentary skeletal muscle. We report the presence of a specific population of nuclear-encoded RNAs in the mitochondria isolated from rat skeletal muscle tissue, which could play a role in regulating exercise adaptations and mitochondrial biology.</p>","PeriodicalId":50455,"journal":{"name":"The FASEB Journal","volume":"39 13","pages":""},"PeriodicalIF":4.4,"publicationDate":"2025-06-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1096/fj.202500157R","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144503225","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":"Therapeutic Potential of GeGen Decoction for the Treatment of Polycystic Ovary Syndrome","authors":"Yiwen Qian,&nbsp;Alamusi Bayoude,&nbsp;Haoyan Zhou,&nbsp;Yuanjiang Shen,&nbsp;Chengzhi Chai","doi":"10.1096/fj.202500192RR","DOIUrl":"https://doi.org/10.1096/fj.202500192RR","url":null,"abstract":"<div>\u0000 \u0000 <p>Polycystic ovary syndrome (PCOS), a prevalent endocrine and metabolic condition during women's reproductive age, is acknowledged as a significant contributory factor for several metabolic disorders. Despite its high prevalence, specific pharmacological interventions for PCOS remain limited. GeGen Decoction (GGD), documented since the Han Dynasty, has been continuously explored for its modern applications. Currently, it is frequently utilized to treat gynecological and metabolic disorders, including PCOS. Our study applied in vivo models utilizing dehydroepiandrosterone (DHEA)-induced PCOS rats and in vitro models employing dihydrotestosterone (DHT) combined with palmitic acid (PA) and oleic acid (OA) to induce KGN cells, to confirm the effects of GGD on PCOS and to explore the mechanism of action. Our findings demonstrated that GGD effectively alleviated PCOS symptoms in rat models by improving estrous cycles, restoring ovarian morphology, regulating hormone levels, and ameliorating glucose and lipid metabolism disorders. Mechanistically, GGD maintained cholesterol homeostasis by modulating the interaction between sterol O-acyltransferase 2 (ACAT2) and insulin-induced gene 1 (INSIG1), thereby enhancing the intracellular environment of KGN cells, promoting the expression of cytochrome P450 family 19 subfamily A member 1 (CYP19A1), and subsequently mitigating PCOS-related symptoms. Our data indicate that GGD may serve as a potential therapeutic agent for PCOS.</p>\u0000 </div>","PeriodicalId":50455,"journal":{"name":"The FASEB Journal","volume":"39 13","pages":""},"PeriodicalIF":4.4,"publicationDate":"2025-06-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144503230","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":"M2-Like Macrophages Exhibit Sialic Acid-Enhanced Efferocytosis via the Siglec CD22","authors":"Emily N. Kukan,&nbsp;Gabrielle L. Fabiano,&nbsp;Leandre M. Glendenning,&nbsp;Julie Y. Zhou,&nbsp;Kevin A. Telfer,&nbsp;James C. Paulson,&nbsp;Brian A. Cobb","doi":"10.1096/fj.202500146RR","DOIUrl":"https://doi.org/10.1096/fj.202500146RR","url":null,"abstract":"<div>\u0000 \u0000 <p>The sialic acid/Siglec axis is an important immunologic regulatory pathway in which host-specific α2,6-sialylated glycans are recognized as markers of self. CD22, known primarily as a surface receptor on B cells, directly prevents autoantigen responses through concurrent recognition of α2,6-linked sialic acids. Here, we report that CD22 is expressed in macrophages polarized to an M2-like, immunomodulatory phenotype. Tissue-resident macrophage populations classically showing an M2-like skew, such as in the lung, were found to be significantly enriched for CD22 expression. We also discovered that CD22 promotes efferocytosis of sialylated glycoproteins and apoptotic debris and is associated with increased protein processing but reduced T cell activation. These findings support a model whereby CD22⁺ M2-like macrophages participate in the resolution of inflammation and a return to tissue homeostasis via the clearance of host-derived α2,6-sialylated debris, degrading this material without further exacerbation of T cell-mediated inflammation.</p>\u0000 </div>","PeriodicalId":50455,"journal":{"name":"The FASEB Journal","volume":"39 13","pages":""},"PeriodicalIF":4.4,"publicationDate":"2025-06-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144503059","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":"Homocysteine: Canary in the Coal Mine or Hidden Threat? A Biochemical Study on the Role of Plasma Thiols","authors":"Daniela Giustarini,&nbsp;Sante Colella,&nbsp;Isabella Dalle-Donne,&nbsp;Ranieri Rossi","doi":"10.1096/fj.202500677RR","DOIUrl":"https://doi.org/10.1096/fj.202500677RR","url":null,"abstract":"<p>Homocysteinemia is routinely measured as a biomarker of cardiovascular risk, but its pathogenic role remains controversial because it is unclear whether—and how—interventions to lower homocysteine levels provide real benefit. In the present original study, we analyzed in detail the effects of oxidative stress, thiol-disulfide exchange reactions, and plasma thiol levels on homocysteinemia. We conducted a clinical study in a group of healthy, homogeneous individuals (<i>n</i> = 62) in which the different redox forms of plasma thiols and several biomarkers of oxidative stress were determined. Homocysteine was characterized by the fact that it was almost completely present as mixed protein disulfide (about 80%–85%). A strong inverse correlation was found between total homocysteine and glutathione concentrations, whereas no correlation was found between homocysteine and oxidative stress markers. The observation that oxidative stress does not affect total homocysteine levels in plasma was confirmed by in vitro treatments of human blood with a special device that allows slow delivery of oxidants. Experiments with cultured cells showed that they can release glutathione in large quantities with different kinetics over time. In addition, a strong inverse correlation between GSH and total homocysteine has been demonstrated in the plasma of humans of different ages and in mammalian species. All these data support the hypothesis that GSH, once released from cells, can trigger a series of thiol-disulfide exchange reactions leading to the cleavage of protein-bound homocysteine and the increase of free homocysteine, thus promoting its excretion. It can therefore be concluded that homocysteinemia can be regulated by the release of GSH from cells and that, consequently, total homocysteine in plasma can be considered a biomarker of cardiovascular risk without necessarily having a direct causal role. The specificity of this process must be taken into account when investigating the pathogenetic role of homocysteine.</p>","PeriodicalId":50455,"journal":{"name":"The FASEB Journal","volume":"39 13","pages":""},"PeriodicalIF":4.4,"publicationDate":"2025-06-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1096/fj.202500677RR","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144503060","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}