{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,13]],"date-time":"2026-10-13T07:01:40Z","timestamp":1791874900437,"version":"4.3.4"},"reference-count":28,"publisher":"Springer Science and Business Media LLC","issue":"1","license":[{"start":{"date-parts":[[2025,12,9]],"date-time":"2025-12-09T00:00:00Z","timestamp":1765238400000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"},{"start":{"date-parts":[[2026,1,13]],"date-time":"2026-01-13T00:00:00Z","timestamp":1768262400000},"content-version":"vor","delay-in-days":35,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"}],"funder":[{"DOI":"10.13039\/501100007128","name":"Natural Science Foundation of Shaanxi Province","doi-asserted-by":"publisher","award":["2021JQ-905"],"award-info":[{"award-number":["2021JQ-905"]}],"id":[{"id":"10.13039\/501100007128","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Key Research and Development Plan of Shaanxi Province","award":["2023-YBSF-478"],"award-info":[{"award-number":["2023-YBSF-478"]}]}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["Sci Rep"],"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>Idiopathic pulmonary arterial hypertension (IPAH) is a progressive and devastating disorder. Mitochondrial oxidative stress has been found to be involved in PAH development. This study aims to explore potential mitochondrial oxidative stress-related biomarkers in IPAH via integrated bioinformatics, validate these genes in MCT-induced rat PAH model and examine the functional roles of COX6B1 and HMGCL in mitochondrial oxidative stress in PDGF-induced pulmonary arterial smooth muscle cells (PASMCs). GSE15197 was downloaded from GEO database. Genes associated with mitochondria and oxidative stress were obtained from Molecular Signatures Database and MitoCarta3.0 databases. Weighted gene co-expression network analysis (WGCNA) was used. GO and KEGG enrichment analysis were utilized. Furthermore, machine learning algorithms (LASSO, SVM-RFE and RF) were constructed to screen hub genes. A nomogram model based on the hub genes were performed for evaluation the diagnostic value, and the expression of hub genes were validated by real-time quantitative PCR and Western blot in MCT-induced rat PAH model. Finally, COX6B1 and HMGCL specific siRNAs were transfected into PDGF-induced PASMCs, mitochondrial ROS (mtROS) level and MnSOD activity were examined. A total of 18 genes related to mitochondrial oxidative stress were identified in IPAH. Two hub genes (COX6B1 and HMGCL) were finally determined as potential biomarkers. Based on the 2 genes, a nomogram model was constructed. This model exhibited remarkable diagnostic efficiency values for IPAH. The expressions of COX6B1 and HMGCL were found to be upregulated in MCT-induced PAH model, in agreement with the findings of bioinformatics analysis. While, transfection with either COX6B1 siRNA or HMGCL siRNA suppressed the abnormal accumulation of mtROS and restored MnSOD activity in PDGF-induced PASMCs. Our results indicated that two mitochondrial oxidative stress-related genes (COX6B1 and HMGCL) were identified as diagnostic biomarkers and may serve as potential targets for IPAH development, although further research is necessary.<\/jats:p>","DOI":"10.1038\/s41598-025-31150-7","type":"journal-article","created":{"date-parts":[[2025,12,9]],"date-time":"2025-12-09T07:05:30Z","timestamp":1765263930000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["Identification of potential biomarkers associated with mitochondrial oxidative stress in idiopathic pulmonary arterial hypertension via bioinformatic and experimental analysis"],"prefix":"10.1038","volume":"16","author":[{"given":"Yanting","family":"Zhu","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiaoming","family":"Wang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiaohui","family":"Yan","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yan","family":"Sun","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Qiong","family":"Wang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Cui","family":"Zhai","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2025,12,9]]},"reference":[{"key":"31150_CR1","doi-asserted-by":"publisher","first-page":"3618","DOI":"10.1093\/eurheartj\/ehac237","volume":"43","author":"M Humbert","year":"2022","unstructured":"Humbert, M. et al. 2022 ESC\/ERS guidelines for the diagnosis and treatment of pulmonary hypertension. Eur. Heart J. 43, 3618\u20133731. https:\/\/doi.org\/10.1093\/eurheartj\/ehac237 (2022).","journal-title":"Eur. Heart J."},{"key":"31150_CR2","doi-asserted-by":"publisher","first-page":"124","DOI":"10.1038\/s41392-024-01839-8","volume":"9","author":"Y Zong","year":"2024","unstructured":"Zong, Y. et al. Mitochondrial dysfunction: Mechanisms and advances in therapy. Signal. Transduct. Target. Ther. 9, 124. https:\/\/doi.org\/10.1038\/s41392-024-01839-8 (2024).","journal-title":"Signal. Transduct. Target. Ther."},{"key":"31150_CR3","doi-asserted-by":"publisher","first-page":"1670","DOI":"10.1111\/bph.13403","volume":"174","author":"A Daiber","year":"2017","unstructured":"Daiber, A. et al. Crosstalk of mitochondria with NADPH oxidase via reactive oxygen and nitrogen species signalling and its role for vascular function. Br. J. Pharmacol. 174, 1670\u20131689. https:\/\/doi.org\/10.1111\/bph.13403 (2017).","journal-title":"Br. J. Pharmacol."},{"key":"31150_CR4","doi-asserted-by":"publisher","first-page":"2601","DOI":"10.1016\/j.cell.2024.04.037","volume":"187","author":"A Suomalainen","year":"2024","unstructured":"Suomalainen, A. & Nunnari, J. Mitochondria at the crossroads of health and disease. Cell 187, 2601\u20132627. https:\/\/doi.org\/10.1016\/j.cell.2024.04.037 (2024).","journal-title":"Cell"},{"key":"31150_CR5","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1038\/s12276-019-0355-7","volume":"51","author":"JN Peoples","year":"2019","unstructured":"Peoples, J. N., Saraf, A., Ghazal, N., Pham, T. T. & Kwong, J. Q. Mitochondrial dysfunction and oxidative stress in heart disease. Exp. Mol. Med. 51, 1\u201313. https:\/\/doi.org\/10.1038\/s12276-019-0355-7 (2019).","journal-title":"Exp. Mol. Med."},{"key":"31150_CR6","doi-asserted-by":"publisher","first-page":"H1235","DOI":"10.1089\/ars.2018.7652","volume":"298","author":"R Rajkumar","year":"2010","unstructured":"Rajkumar, R. et al. Genomewide RNA expression profiling in lung identifies distinct signatures in idiopathic pulmonary arterial hypertension and secondary pulmonary hypertension. Am. J. Physiol. Heart Circ. Physiol. 298, H1235\u20131248. https:\/\/doi.org\/10.1089\/ars.2018.7652 (2010).","journal-title":"Am. J. Physiol. Heart Circ. Physiol."},{"key":"31150_CR7","doi-asserted-by":"publisher","first-page":"2361","DOI":"10.1056\/NEJMra2000348","volume":"385","author":"PM Hassoun","year":"2021","unstructured":"Hassoun, P. M. Pulmonary arterial hypertension. N Engl. J. Med. 385, 2361\u20132376. https:\/\/doi.org\/10.1056\/NEJMra2000348 (2021).","journal-title":"N Engl. J. Med."},{"key":"31150_CR8","doi-asserted-by":"publisher","first-page":"129","DOI":"10.1111\/joim.12623","volume":"282","author":"P Ghataorhe","year":"2017","unstructured":"Ghataorhe, P. et al. Pulmonary arterial hypertension - progress in Understanding the disease and prioritizing strategies for drug development. J. Intern. Med. 282, 129\u2013141. https:\/\/doi.org\/10.1111\/joim.12623 (2017).","journal-title":"J. Intern. Med."},{"key":"31150_CR9","doi-asserted-by":"publisher","first-page":"310","DOI":"10.1016\/j.healun.2019.12.012","volume":"39","author":"R Badagliacca","year":"2020","unstructured":"Badagliacca, R. et al. Clinical implications of idiopathic pulmonary arterial hypertension phenotypes defined by cluster analysis. J. Heart Lung Transpl. 39, 310\u2013320. https:\/\/doi.org\/10.1016\/j.healun.2019.12.012 (2020).","journal-title":"J. Heart Lung Transpl."},{"key":"31150_CR10","doi-asserted-by":"publisher","first-page":"447","DOI":"10.1089\/ars.2018.7652","volume":"32","author":"Z Yang","year":"2020","unstructured":"Yang, Z. et al. Important role of sarcoplasmic reticulum Ca(2+) release via Ryanodine Receptor-2 channel in hypoxia-induced rieske iron-sulfur protein-mediated mitochondrial reactive oxygen species generation in pulmonary artery smooth muscle cells. Antioxid. Redox Signal. 32, 447\u2013462. https:\/\/doi.org\/10.1089\/ars.2018.7652 (2020).","journal-title":"Antioxid. Redox Signal."},{"key":"31150_CR11","doi-asserted-by":"publisher","first-page":"1","DOI":"10.3390\/antiox12122060","volume":"12","author":"S Yan","year":"2023","unstructured":"Yan, S., Sheak, J. R., Walker, B. R., Jernigan, N. L. & Resta, T. C. Contribution of mitochondrial reactive oxygen species to chronic hypoxia-induced pulmonary hypertension. Antioxid. (Basel) 12, 1\u201323. https:\/\/doi.org\/10.3390\/antiox12122060 (2023).","journal-title":"Antioxid. (Basel)"},{"key":"31150_CR12","doi-asserted-by":"publisher","first-page":"9","DOI":"10.1016\/j.freeradbiomed.2022.09.026","volume":"193","author":"J Sevilla-Montero","year":"2022","unstructured":"Sevilla-Montero, J. et al. Cigarette smoke induces pulmonary arterial dysfunction through an imbalance in the redox status of the soluble Guanylyl cyclase. Free Radic Biol. Med. 193, 9\u201322. https:\/\/doi.org\/10.1016\/j.freeradbiomed.2022.09.026 (2022).","journal-title":"Free Radic Biol. Med."},{"key":"31150_CR13","doi-asserted-by":"publisher","first-page":"2078","DOI":"10.1093\/hmg\/ddt601","volume":"23","author":"KK Kemppainen","year":"2014","unstructured":"Kemppainen, K. K. et al. Expression of alternative oxidase in drosophila ameliorates diverse phenotypes due to cytochrome oxidase deficiency. Hum. Mol. Genet. 23, 2078\u20132093. https:\/\/doi.org\/10.1093\/hmg\/ddt601 (2014).","journal-title":"Hum. Mol. Genet."},{"key":"31150_CR14","doi-asserted-by":"publisher","first-page":"159","DOI":"10.1038\/ejhg.2014.85","volume":"23","author":"UN Abdulhag","year":"2015","unstructured":"Abdulhag, U. N. et al. Mitochondrial complex IV deficiency, caused by mutated COX6B1, is associated with encephalomyopathy, hydrocephalus and cardiomyopathy. Eur. J. Hum. Genet. 23, 159\u2013164. https:\/\/doi.org\/10.1038\/ejhg.2014.85 (2015).","journal-title":"Eur. J. Hum. Genet."},{"key":"31150_CR15","doi-asserted-by":"publisher","first-page":"9787","DOI":"10.1007\/s11357-015-9787-8","volume":"37","author":"SE Kim","year":"2015","unstructured":"Kim, S. E. et al. Upregulation of cytochrome c oxidase subunit 6b1 (Cox6b1) and formation of mitochondrial supercomplexes: Implication of Cox6b1 in the effect of calorie restriction. Age (Dordr) 37, 9787. https:\/\/doi.org\/10.1007\/s11357-015-9787-8 (2015).","journal-title":"Age (Dordr)"},{"key":"31150_CR16","doi-asserted-by":"publisher","first-page":"59","DOI":"10.1007\/s10529-018-2614-4","volume":"41","author":"W Zhang","year":"2019","unstructured":"Zhang, W., Wang, Y., Wan, J., Zhang, P. & Pei, F. COX6B1 relieves hypoxia\/reoxygenation injury of neonatal rat cardiomyocytes by regulating mitochondrial function. Biotechnol. Lett. 41, 59\u201368. https:\/\/doi.org\/10.1007\/s10529-018-2614-4 (2019).","journal-title":"Biotechnol. Lett."},{"key":"31150_CR17","doi-asserted-by":"publisher","first-page":"4852","DOI":"10.3892\/mmr.2019.10144","volume":"19","author":"S Yang","year":"2019","unstructured":"Yang, S., Wu, P., Xiao, J. & Jiang, L. Overexpression of COX6B1 protects against I\/R\u2013induced neuronal injury in rat hippocampal neurons. Mol. Med. Rep. 19, 4852\u20134862. https:\/\/doi.org\/10.3892\/mmr.2019.10144 (2019).","journal-title":"Mol. Med. Rep."},{"key":"31150_CR18","doi-asserted-by":"publisher","first-page":"33227","DOI":"10.1074\/jbc.M112.393231","volume":"287","author":"C Montgomery","year":"2012","unstructured":"Montgomery, C., Pei, Z., Watkins, P. A. & Miziorko, H. M. Identification and characterization of an extramitochondrial human 3-hydroxy-3-methylglutaryl-CoA lyase. J. Biol. Chem. 287, 33227\u201333236. https:\/\/doi.org\/10.1074\/jbc.M112.393231 (2012).","journal-title":"J. Biol. Chem."},{"key":"31150_CR19","doi-asserted-by":"publisher","first-page":"e110466","DOI":"10.15252\/embj.2021110466","volume":"41","author":"V Gouirand","year":"2022","unstructured":"Gouirand, V. et al. Ketogenic HMG-CoA lyase and its product \u03b2-hydroxybutyrate promote pancreatic cancer progression. Embo j. 41, e110466. https:\/\/doi.org\/10.15252\/embj.2021110466 (2022).","journal-title":"Embo j."},{"key":"31150_CR20","doi-asserted-by":"publisher","first-page":"11954","DOI":"10.1038\/s41598-017-11025-2","volume":"7","author":"W Luo","year":"2017","unstructured":"Luo, W. et al. Inactivation of HMGCL promotes proliferation and metastasis of nasopharyngeal carcinoma by suppressing oxidative stress. Sci. Rep. 7, 11954. https:\/\/doi.org\/10.1038\/s41598-017-11025-2 (2017).","journal-title":"Sci. Rep."},{"key":"31150_CR21","doi-asserted-by":"publisher","first-page":"753004","DOI":"10.3389\/fcell.2021.753004","volume":"9","author":"J Liu","year":"2021","unstructured":"Liu, J. et al. Development of an oxidative Phosphorylation-Related and immune microenvironment prognostic signature in uterine corpus endometrial carcinoma. Front. Cell. Dev. Biol. 9, 753004. https:\/\/doi.org\/10.3389\/fcell.2021.753004 (2021).","journal-title":"Front. Cell. Dev. Biol."},{"key":"31150_CR22","doi-asserted-by":"publisher","first-page":"1498","DOI":"10.12122\/j.issn.1673-4254.2025.07.16","volume":"45","author":"X Gong","year":"2025","unstructured":"Gong, X. et al. [LncRNA SNHG15 promotes proliferation, migration and invasion of lung adenocarcinoma cells by regulating COX6B1 through sponge adsorption of miR-30b-3p]. Nan Fang Yi Ke Da Xue Xue Bao 45, 1498\u20131505. https:\/\/doi.org\/10.12122\/j.issn.1673-4254.2025.07.16 (2025).","journal-title":"Nan Fang Yi Ke Da Xue Xue Bao"},{"key":"31150_CR23","doi-asserted-by":"publisher","first-page":"653","DOI":"10.1093\/neuonc\/noad232","volume":"26","author":"Y Sun","year":"2024","unstructured":"Sun, Y. et al. Metabolic modulation of histone acetylation mediated by HMGCL activates the FOXM1\/\u03b2-catenin pathway in glioblastoma. Neuro Oncol. 26, 653\u2013669. https:\/\/doi.org\/10.1093\/neuonc\/noad232 (2024).","journal-title":"Neuro Oncol."},{"key":"31150_CR24","doi-asserted-by":"publisher","first-page":"27","DOI":"10.1093\/nar\/28.1.27","volume":"28","author":"M Kanehisa","year":"2000","unstructured":"Kanehisa, M. & Goto, S. KEGG: Kyoto encyclopedia of genes and genomes. Nucleic Acids Res. 28, 27\u201330. https:\/\/doi.org\/10.1093\/nar\/28.1.27 (2000).","journal-title":"Nucleic Acids Res."},{"key":"31150_CR25","doi-asserted-by":"publisher","first-page":"1947","DOI":"10.1002\/pro.3715","volume":"28","author":"M Kanehisa","year":"2019","unstructured":"Kanehisa, M. Toward understanding the origin and evolution of cellular organisms. Protein Sci. 28, 1947\u20131951. https:\/\/doi.org\/10.1002\/pro.3715 (2019).","journal-title":"Protein Sci."},{"key":"31150_CR26","doi-asserted-by":"publisher","first-page":"D587","DOI":"10.1093\/nar\/gkac963","volume":"51","author":"M Kanehisa","year":"2023","unstructured":"Kanehisa, M., Furumichi, M., Sato, Y., Kawashima, M. & Ishiguro-Watanabe, M. KEGG for taxonomy-based analysis of pathways and genomes. Nucleic Acids Res. 51, D587\u2013d592. https:\/\/doi.org\/10.1093\/nar\/gkac963 (2023).","journal-title":"Nucleic Acids Res."},{"key":"31150_CR27","doi-asserted-by":"publisher","first-page":"535","DOI":"10.1186\/s12872-024-04192-4","volume":"24","author":"RJ Gao","year":"2024","unstructured":"Gao, R. J. et al. Quercetin regulates pulmonary vascular remodeling in pulmonary hypertension by downregulating TGF-\u03b21-Smad2\/3 pathway. BMC Cardiovasc. Disord 24, 535. https:\/\/doi.org\/10.1186\/s12872-024-04192-4 (2024).","journal-title":"BMC Cardiovasc. Disord"},{"key":"31150_CR28","doi-asserted-by":"publisher","first-page":"208","DOI":"10.1186\/s10020-024-00975-9","volume":"30","author":"C Chen","year":"2024","unstructured":"Chen, C. et al. PI3K p85\u03b1\/HIF-1\u03b1 accelerates the development of pulmonary arterial hypertension by regulating fatty acid uptake and mitophagy. Mol. Med. 30, 208. https:\/\/doi.org\/10.1186\/s10020-024-00975-9 (2024).","journal-title":"Mol. Med."}],"container-title":["Scientific Reports"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.nature.com\/articles\/s41598-025-31150-7.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/s41598-025-31150-7","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/s41598-025-31150-7.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,1,15]],"date-time":"2026-01-15T10:49:47Z","timestamp":1768474187000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.nature.com\/articles\/s41598-025-31150-7"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,12,9]]},"references-count":28,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2026,12]]}},"alternative-id":["31150"],"URL":"https:\/\/doi.org\/10.1038\/s41598-025-31150-7","relation":{},"ISSN":["2045-2322"],"issn-type":[{"value":"2045-2322","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,12,9]]},"assertion":[{"value":"15 June 2025","order":1,"name":"received","label":"Received","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"30 November 2025","order":2,"name":"accepted","label":"Accepted","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"9 December 2025","order":3,"name":"first_online","label":"First Online","group":{"name":"ArticleHistory","label":"Article History"}},{"order":1,"name":"Ethics","group":{"name":"EthicsHeading","label":"Declarations"}},{"value":"The authors declare no competing interests.","order":2,"name":"Ethics","group":{"name":"EthicsHeading","label":"Competing interests"}},{"value":"All animal care and experiments were performed in accordance with the Guide for the Care and Use of Laboratory Animals of Xi\u2019an Medical University Animal Experiment Center. All protocols used in this study were approved by the Laboratory Animal Care Committee of Xi\u2019an Medical University. The study was carried out in compliance with the ARRIVE guidelines.","order":3,"name":"Ethics","group":{"name":"EthicsHeading","label":"Ethical approval"}}],"article-number":"1647"}}