Journal of Bioenergetics and Biomembranes最新文献

{"title":"Protonophore uncouplers and the decoupler α,ω-hexadecanedioic acid relieve the succinate-induced blockade of glutamate and malate oxidation in the resting state (state 4).","authors":"Svetlana I Pavlova, Victor N Samartsev, Elya I Kuzmina, Galina I Zainutdinova, Mikhail V Dubinin","doi":"10.1007/s10863-026-10129-7","DOIUrl":"10.1007/s10863-026-10129-7","url":null,"abstract":"<p><p>Theoretical and experimental studies have revealed that in liver mitochondria in the absence of ATP synthesis (state 4), the oxidation of a mixture of succinate with glutamate and malate, like the oxidation of succinate alone, involves only complexes III and IV in the generation of the proton motive force, while complex II, which does not pump protons, oxidizes succinate. Consequently, succinate oxidation suppresses the oxidation of glutamate and malate, likely by inducing reverse electron transport (RET) at complex I and inhibiting NADH oxidation. The concentration of succinate at which its half-maximal effect on blocking glutamate and malate oxidation is observed is 220 ± 25 µM, which corresponds to its concentration in cells of various organs and tissues under physiological conditions. The protonophore uncouplers n-trifluoromethoxycarbonylcyanide phenylhydrazone (FCCP), palmitic acid (PA), and chenodeoxycholic acid (CDCA), provided that they stimulate respiration no more than two-fold alleviate the suppression of glutamate and malate oxidation by succinate. The decoupler α,ω-hexadecanedioic acid (HDA), upon stimulating mitochondrial respiration by 1.5-fold, also alleviates the suppressive effect of succinate on glutamate and malate oxidation. It is noted that, unlike protonophore uncouplers, this effect of HDA may be associated with switching complex III of the respiratory chain to an idle mode of operation. During the oxidation of glutamate and malate, i.e., under conditions of forward electron transport, the protonophore uncouplers FCCP, PA, and CDCA inhibit H₂O₂ generation by liver mitochondria, whereas HDA is ineffective. During the oxidation of succinate and the mixture of succinate with glutamate and malate, i.e., under conditions of reverse electron transport (RET), protonophore uncouplers and HDA inhibit H₂O₂ generation by liver mitochondria to approximately the same extent. It is suggested that this effect of the aforementioned uncouplers and HDA is likely related to the inhibition of RET and, consequently, to the alleviation of the suppressive effect of succinate on glutamate and malate oxidation. This study demonstrates that the physiological concentration of succinate is sufficient to dominate electron flow, and that both classical and natural uncouplers, as well as the decoupler HDA, can reverse this effect, albeit via different mechanistic pathways.</p>","PeriodicalId":15080,"journal":{"name":"Journal of Bioenergetics and Biomembranes","volume":"58 1","pages":""},"PeriodicalIF":3.7,"publicationDate":"2026-08-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148828632","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"M2 macrophage-derived exosomal EHF transcriptionally activates FGFR1 to promote malignant phenotypes and glycolysis in NSCLC.","authors":"Dan Zhang, Wei Wu, Peng Ge, Xia Cao, Yuan Qin, Danfeng Dong, Jin Yang","doi":"10.1007/s10863-026-10121-1","DOIUrl":"10.1007/s10863-026-10121-1","url":null,"abstract":"&lt;p&gt;&lt;strong&gt;Background: &lt;/strong&gt;Non-small cell lung cancer (NSCLC) remains a leading cause of cancer-related mortality worldwide, largely due to the supportive role of the tumor microenvironment (TME). Tumor-associated macrophages, particularly the M2 phenotype, are pivotal in promoting NSCLC progression. Exosomes, key mediators of intercellular communication, can transfer functional cargo from M2 macrophages to cancer cells, thereby regulating malignant behaviors. However, the specific mechanisms by which M2 macrophage-derived exosomes modulate NSCLC progression are not fully understood.&lt;/p&gt;&lt;p&gt;&lt;strong&gt;Methods: &lt;/strong&gt;Bioinformatics analyses were initially performed to identify differentially expressed genes (DEGs) in M2 macrophages and NSCLC tissues using the GSE159112 and GSE268175 datasets. THP-1 cells were induced to differentiate into M2 macrophages, from which exosomes were isolated and characterized via nanoparticle tracking analysis, transmission electron microscopy, and western blotting. NSCLC cell lines (A549 and H23) were co-cultured with these exosomes to assess the transfer of ETS homologous factor (EHF). Functional assays, including 5-Ethynyl-2'-deoxyuridine (EdU), Transwell, flow cytometry, and sphere formation assays, were conducted to evaluate cell proliferation, migration, invasion, apoptosis, and stemness. Glycolytic capacity was determined by measuring glucose uptake, lactate production, and ATP levels. Chromatin immunoprecipitation (ChIP) and dual-luciferase reporter assays were employed to verify the transcriptional regulation of FGFR1 by EHF. Rescue experiments involving FGFR1 overexpression were performed to validate the signaling axis, and a xenograft mouse model was established to confirm the in vivo findings.&lt;/p&gt;&lt;p&gt;&lt;strong&gt;Results: &lt;/strong&gt;EHF was identified as a critical upregulated transcription factor in both M2 macrophages and NSCLC tissues. M2 macrophage-derived exosomes were efficiently internalized by NSCLC cells, resulting in subsequent upregulation of its expression in recipient cells. Mechanistically, EHF transcriptionally activated fibroblast growth factor receptor 1 (FGFR1) by binding to its promoter region. Functionally, exosomes derived from EHF-deficient M2 macrophages significantly suppressed NSCLC cell proliferation, migration, invasion, and sphere formation, while promoting apoptosis. Concurrently, the loss of exosomal EHF inhibited glycolysis, evidenced by reduced glucose uptake, lactate production, and ATP levels. Crucially, restoring FGFR1 expression reversed the suppressive effects induced by EHF-deficient exosomes, confirming that the EHF/FGFR1 axis drives these malignant phenotypes. In vivo experiments further demonstrated that exosomes from EHF-silenced M2 macrophages inhibited tumor growth and downregulated proliferation markers.&lt;/p&gt;&lt;p&gt;&lt;strong&gt;Conclusion: &lt;/strong&gt;M2 macrophage-derived exosomal EHF promoted NSCLC progression and glycolysis by transcriptionally activating FGFR1. These finding","PeriodicalId":15080,"journal":{"name":"Journal of Bioenergetics and Biomembranes","volume":"58 1","pages":""},"PeriodicalIF":3.7,"publicationDate":"2026-08-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148706586","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Fluoride exposure induces oxidative damage in primary astrocytes of SD rats and changes in the expression of α-synuclein and its effect on Nrf2-NQO1/HO-1 signaling pathway.","authors":"Xiao Zhou, Kaiju Mo, Wen Wan, Ling Ye, Minghai Liu, Zhizhong Guan, Yuchuan Ding, Yanjie Liu, Shu Xu","doi":"10.1007/s10863-026-10128-8","DOIUrl":"10.1007/s10863-026-10128-8","url":null,"abstract":"<p><p>Long-term excessive fluoride intake accumulates in brain tissue, causing neuronal degeneration and nervous system dysfunction. This mechanism is closely related to elevated oxidative stress. α-Synuclein (α-Syn) is expressed in neuronal presynaptic terminals; under environmental toxin exposure or oxidative stress, its expression changes and it misfolds into oligomers and aggregates that are cytotoxic and induce oxidative cellular damage. Meanwhile, the nuclear factor erythroid 2-related factor 2 (Nrf2)-NAD(P)H: quinone oxidoreductase 1 (NQO1)/heme oxygenase-1 (HO-1) pathway is an important cellular system for regulating oxidative damage, and its core kinase is mainly expressed in brain astrocytes.The aim of this study is to investigate the effects of fluoride exposure on oxidative stress levels in astrocytes and the expression of α-Syn within the Nrf2-NQO1/HO-1 pathway. Primary astrocytes were treated with extracellular α-synuclein pre-formed fibrils (αSP) in conjunction with fluoride exposure. The results demonstrated that fluoride exposure induced reactive activation of primary astrocytes, increased levels of reactive oxygen species (ROS), decreased activities of superoxide dismutase (SOD) and catalase (CAT), and up-regulated the protein expression within the Nrf2-NQO1/HO-1 pathway. αSP caused oxidative stress in primary astrocytes and exacerbated oxidative damage induced by fluoride exposure. These findings are significant for enhancing the understanding of the mechanisms underlying brain injury associated with chronic fluorosis.</p>","PeriodicalId":15080,"journal":{"name":"Journal of Bioenergetics and Biomembranes","volume":"58 1","pages":""},"PeriodicalIF":3.7,"publicationDate":"2026-08-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148684557","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Aloe vera extract disrupts mitochondrial bioenergetics and suppresses HeLa cervical cancer cell proliferation: Transcriptomic and metabolic insights into cisplatin co-treatment.","authors":"Tuğba Kul Köprülü, Burçin Erkal Çam","doi":"10.1007/s10863-026-10127-9","DOIUrl":"10.1007/s10863-026-10127-9","url":null,"abstract":"<p><p>Cervical cancer remains a leading cause of cancer related mortality in women worldwide. Cisplatin (CP) is a cornerstone chemotherapeutic agent; however, its clinical utility is constrained by dose-limiting toxicity and the emergence of drug resistance, necessitating novel combinatorial strategies. This study investigated the therapeutic potential of combining CP with Aloe vera extract (AVE) against HeLa cervical cancer cells and the normal human skin fibroblast cell line CCD-1072Sk. Cytotoxicity assays performed using the xCELLigence Real-Time Cell Analysis (RTCA) system determined IC₅₀ values of 18.45 µM for CP and 18.88 µg/mL for AVE in HeLa cells, compared with 53.31 µM and 19.09 µg/mL, respectively, in CCD-1072Sk cells. The CP + AVE combination exhibited markedly enhanced antiproliferative activity in HeLa cells compared with CP alone, while CCD-1072Sk cells exhibited continued but decelerated proliferation, suggesting a differential cellular response. Seahorse metabolic analysis revealed that AVE, both alone and in combination with CP, markedly increased mitochondrial oxygen consumption rate (OCR) and ATP-linked respiration in HeLa cells, indicating induction of sustained bioenergetic stress beyond the cells' adaptive capacity. Transcriptomic profiling of HeLa cells identified 453 differentially expressed genes (DEGs) upon combination treatment, with significant downregulation of oxidative phosphorylation (OXPHOS) components, mitochondrial electron transport chain subunits, and the fatty acid metabolism gene CPT1B, collectively indicating a severe disruption of cellular energy metabolism. DepMap database analysis confirmed that downregulated targets including CPT1B and CHN2 are functionally important for HeLa cell survival. Collectively, these data suggest that AVE drives HeLa cells into a hypermetabolic state that overloads mitochondrial capacity; in combination with CP, this converging bioenergetic stress produces enhanced cytotoxicity descriptively consistent with an additive interaction (a term used here in its dose-response sense, as no formal combination-index or Bliss/Loewe synergy analysis was performed) at the IC₅₀ doses employed. This study highlights the potential of AVE as a metabolically active combinatorial agent capable of potentiating CP efficacy by targeting cancer cell energy metabolism, and provides a mechanistic basis for further investigation.</p>","PeriodicalId":15080,"journal":{"name":"Journal of Bioenergetics and Biomembranes","volume":"58 1","pages":""},"PeriodicalIF":3.7,"publicationDate":"2026-08-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148678756","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Correction: Succinate and lactate produced as conserved biomarkers through chronic and transient substrate-level phosphorylation: from microorganisms to cancer.","authors":"Derek C Lee, Tomas Duraj, Christos Chinopoulos, Thomas N Seyfried","doi":"10.1007/s10863-026-10124-y","DOIUrl":"10.1007/s10863-026-10124-y","url":null,"abstract":"","PeriodicalId":15080,"journal":{"name":"Journal of Bioenergetics and Biomembranes","volume":"58 1","pages":""},"PeriodicalIF":3.7,"publicationDate":"2026-08-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13437563/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148669443","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Par-4/TERT induced autophagy dysregulation in regulating glycolysis in hyperglycemia combined with hepatocellular carcinoma.","authors":"Jiaying Tang, Ping Wang, Xiangfen Chen, Xiaotian Lei, Sheng Qiu, Qinan Wu","doi":"10.1007/s10863-026-10126-w","DOIUrl":"10.1007/s10863-026-10126-w","url":null,"abstract":"<p><p>To explore the mechanism by which the Prostate apoptosis response-4 (Par-4)/Telomerase Reverse Transcriptase (TERT) axis induces tumor progression by regulating autophagy and glycolysis in hepatocellular carcinoma (HCC) under hyperglycemic conditions. The high-glucose Hepa1-6 cell model and nude mouse HCC xenograft model were used, along with gene knockdown (shPar-4, shTERT) and overexpression (oePar-4) strategies. Methylthiazolyldiphenyl-tetrazolium bromide (MTT) assay, flow cytometry, Western blot, TUNEL staining, co-immunoprecipitation (co-IP), quantitative real-time PCR (qPCR), and animal experiments were performed to analyze the effects of Par-4/TERT on AKT activity, autophagic activity, and the expression of key glycolytic enzymes (GLUT-1, HK, PFK, and PK). The anti-tumor effects of AKT inhibitors (SH-5 and LY294002), the autophagy regulator (rapamycin), and the glycolysis inhibitor (2-deoxy-D-glucose, 2-DG) were further evaluated. Under hyperglycemic conditions, Par-4 expression was significantly suppressed, whereas TERT expression was markedly upregulated. This elevated TERT expression activated the AKT/NF-κB/FOXO3a signaling pathway, leading to enhanced glycolytic metabolism (evidenced by upregulated expression of glucose transporters such as GLUT-1 and key glycolytic enzymes) and induced autophagic processes (characterized by increased LC3-II/LC3-I ratios and decreased p62 levels). Par-4 overexpression or TERT knockdown effectively reversed these effects, resulting in inhibited cell proliferation and induced apoptosis. Co-immunoprecipitation (co-IP) experiments confirmed a direct interaction between Par-4 and TERT, with Par-4 negatively regulating TERT expression. In combination therapeutic approaches, the LY294002 + 2-DG regimen exhibited synergistic anti-tumor efficacy in models with Par-4 overexpression or TERT knockdown, achieving a greater than 50% reduction in tumor volume accompanied by downregulated p62 expression. The Par-4/TERT regulatory axis modulates autophagy dysregulation and glycolytic metabolism through the AKT signaling cascade, contributing to HCC progression under hyperglycemic conditions. Synergistic targeting of this axis, in conjunction with AKT blockade and glycolytic inhibition, exerts potent anti-tumor activity, providing a novel precision therapeutic strategy for HCC associated with hyperglycemia.</p>","PeriodicalId":15080,"journal":{"name":"Journal of Bioenergetics and Biomembranes","volume":"58 1","pages":""},"PeriodicalIF":3.7,"publicationDate":"2026-07-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148630625","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Application of glutamine metabolism-related gene signatures in the non-invasive diagnosis of coronary artery disease: integrating bulk and single-cell transcriptomics.","authors":"Zhiyong Zhao, Yu Li, Junpeng Li, Yongqian Gao, Zhijun Yan, Hui Li, Qiuli Liu","doi":"10.1007/s10863-026-10125-x","DOIUrl":"10.1007/s10863-026-10125-x","url":null,"abstract":"<p><p>Coronary artery disease (CAD) is a common cardiovascular disorder strongly associated with glutamine metabolism. This study seeks to identify novel glutamine metabolism-related gene markers in CAD. Based on GEO datasets and glutamine metabolism-related genes, hub genes were pinpointed through WGCNA combined with LASSO, SVM-RFE, and random forest algorithms. Immune cell infiltration was estimated with the CIBERSORT algorithm. To confirm the expression and roles of hub genes, ox-LDL-stimulated HUVECs were analyzed in culture. MYBPC3 and TRIM47 emerged as candidate diagnostic signatures for CAD. Associations between MYBPC3/TRIM47 and immune cell infiltration were revealed. Upregulation of MYBPC3 and TRIM47 in CAD was confirmed in both GSE113079 and ox-LDL-treated HUVECs. Knockdown of TRIM47 or MYBPC3 promoted ox-LDL-induced proliferation of HUVECs, alleviated cellular apoptosis, and suppressed the expression of pro-inflammatory cytokines (IL-1β, IL-6, TNF-α) as well as M1 macrophage polarization. MYBPC3 and TRIM47 may serve as candidate diagnostic signatures for CAD.</p>","PeriodicalId":15080,"journal":{"name":"Journal of Bioenergetics and Biomembranes","volume":"58 1","pages":""},"PeriodicalIF":3.7,"publicationDate":"2026-07-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148619794","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"The fullerenol C₆₀(OH)₃₆ influence on the structural-dynamic and functional parameters of mitochondria.","authors":"Ashura Israfilova, Albina Dzhafarova, Vagab Abdulaev, Aina Adieva, Maria Markova, Kamila Sfieva, Anna Titova, Zhanna Lyutova, Alina Borisenkova","doi":"10.1007/s10863-026-10123-z","DOIUrl":"10.1007/s10863-026-10123-z","url":null,"abstract":"<p><p>Fullerenols, which possess antioxidants, anti-inflammatory, and neuroprotective properties, can affect mitochondria. The effect of fullerenol C₆₀(OH)₃₆ on the structural-dynamic and functional parameters of mitochondria, both isolated in vitro and in vivo in Wistar rats, was studied. It was found that fullerenol at a dose of 10 mg/kg reduced bioenergetic parameters of both glutamate-dependent and succinate-dependent intracellular respiration in vivo. In the in vitro model system, fullerenol at a dose of 0.2 mg/ml significantly decreased the oxygen consumption rates in all metabolic states, leading to a reduction in the Lardy and Chance respiratory control ratios and the P/O ratio. Conversely, at a dose of 0.1 µg/ml, fullerenols significantly increased these respiratory parameters. Fullerenols, both in vivo and in vitro, increased the viscosity of annular and total mitochondrial membrane lipids. They did not affect the polarity of total lipids but decreased the polarity of annular lipids. Fullerenols at high doses induced significant conformational changes in membrane proteins, increasing their immersion into the lipid matrix. Our results demonstrate that fullerenols modulate mitochondrial bioenergetics and membrane structural dynamics in a dose-dependent manner, which may have implications for understanding both the cytoprotective and potentially cytotoxic effects of these nanoparticles.</p>","PeriodicalId":15080,"journal":{"name":"Journal of Bioenergetics and Biomembranes","volume":"58 1","pages":""},"PeriodicalIF":3.7,"publicationDate":"2026-07-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148603388","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"N6 methyl adenosine-stabilized microtubule actin crosslinking factor 1 ameliorates staphylococcal protein A-induced damage in osteocyte-like MLO-Y4 cells.","authors":"Siyu Lu, Muguo Song, Lirong Ren, Junyi Li, Kehan Lv, Jian Shi, Yongqing Xu","doi":"10.1007/s10863-026-10122-0","DOIUrl":"10.1007/s10863-026-10122-0","url":null,"abstract":"<p><p>Staphylococcus aureus-induced bone infection is one of the thorny issues in clinical orthopedics, and understanding its detailed molecular regulatory mechanisms will help in seeking strategies. In this study, the differentially expressed genes in murine long bone osteocyte (MLO)-Y4 cells treated with or without Staphylococcus aureus were screened out through RNA sequencing, and microtubule actin crosslinking factor 1 (MACF1) was identified as one of the most significantly downregulated genes, which was validated by the subsequent RT-qPCR and Western blot. Then, Cellular functional assays showed that MACF1 overexpression markedly inhibited apoptosis and secretion of inflammatory mediators, but recovered cell viability and production of osteogenic markers in staphylococcal protein A (SPA) induced MLO-Y4 cells. Mechanistically, MACF1 mRNA was stabilized by N6 methyl adenosine (m6A) modification, and upregulated MACF1 inactivated the PI3K/AKT signaling in SPA-induced MLO-Y4 cells. To conclude, targeting m6A-methylated MACF1 ameliorates SPA-triggered inflammation, apoptosis and metabolic dysfunction in MLO-Y4 cells.</p>","PeriodicalId":15080,"journal":{"name":"Journal of Bioenergetics and Biomembranes","volume":"58 1","pages":""},"PeriodicalIF":3.7,"publicationDate":"2026-07-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148603355","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Methotrexate potentiates oxidative stress-mediated transdifferentiation and metabolic reprogramming of neutrophils.","authors":"Aniruddha Bagchi, Shilpa Sengupta, Parasar Ghosh, Alakendu Ghosh, Mitali Chatterjee","doi":"10.1007/s10863-026-10119-9","DOIUrl":"10.1007/s10863-026-10119-9","url":null,"abstract":"<p><p>Rheumatoid arthritis (RA) is associated with inflammation, oxidative stress and infiltration of immune cells (~ 90% neutrophils) into inflamed joints. Neutrophils have demonstrated the ability to transdifferentiate into 'neutrophil-dendritic cell hybrids' or N-DCs, acquire antigen presenting properties (HLA-DR/CD80/CD86) and demonstrate enhanced oxidative stress. Classical neutrophils rely mainly on glycolysis, but the bioenergetics of N-DCs remains poorly defined. Accordingly, this study aimed to assess the impact of methotrexate (MTX) on neutrophils with regard to their transdifferentiation and bioenergetics. The ex-vivo effect of MTX on neutrophils sourced from healthy controls was studied (+/- Phorbol 12-myristate 13-acetate, PMA) in terms of CD83⁺, HLA-DR⁺, generation of reactive oxygen species, activation (myeloperoxidase) and apoptotic status (Annexin V, Caspase 3 and Bcl-2) by flow cytometry, while the bioenergetics was assessed using Agilent XFp analyser, along with expression of key metabolic regulatory enzymes by real time PCR. In neutrophils, MTX did not impact on the generation of ROS; however, in a pro-oxidant milieu induced by PMA, MTX demonstrated a synergistic enhancement in the generation of ROS, increased %CD83^high/HLA-DR^high, raised myeloperoxidase, elevation of apoptotic markers (Annexin V, Caspase 3) along with downregulation of Bcl-2. MTX per se did not alter the oxygen consumption rate (OCR), whereas following PMA-induced oxidative stress, MTX enhanced OCR, while glycolytic mechanisms remained unaltered. MTX, the gold standard for treatment of RA did not alter the redox status of neutrophils, but in a PMA-induced pro-oxidant milieu, MTX facilitated transdifferentiation and activation of neutrophils, altered their apoptotic and bioenergetic status, suggestive of an adverse bystander effect.</p>","PeriodicalId":15080,"journal":{"name":"Journal of Bioenergetics and Biomembranes","volume":"58 1","pages":""},"PeriodicalIF":3.7,"publicationDate":"2026-07-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148536076","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}