{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,2]],"date-time":"2026-07-02T18:51:59Z","timestamp":1783018319935,"version":"3.54.6"},"reference-count":32,"publisher":"Public Library of Science (PLoS)","issue":"7","license":[{"start":{"date-parts":[[2026,7,2]],"date-time":"2026-07-02T00:00:00Z","timestamp":1782950400000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["www.plosone.org"],"crossmark-restriction":false},"short-container-title":["PLoS One"],"abstract":"<jats:p>Simulated diving and decompression can impair endothelial function, but the upstream oxidant sources and their relationship with endothelial nitric oxide synthase (eNOS) coupling in the pulmonary circulation remain unclear. We investigated whether NADPH oxidase 2 (NOX2) is associated with oxidative stress, tetrahydrobiopterin (BH4) depletion, altered eNOS coupling, and pulmonary endothelial dysfunction after simulated air diving. Eighteen male Sprague\u2013Dawley rats were assigned to three groups: control, decompression stress, and decompression stress with the NOX2 inhibitor GSK2795039 (100 mg\/kg, intraperitoneal) administered before pressurization. Decompression stress was induced by hyperbaric exposure to 600 kPa for 1 h followed by decompression to ambient pressure; pulmonary arteries were collected 1 h after decompression. We evaluated NOX2 expression, oxidative stress indices, BH4 content, eNOS phosphorylation and dimer\/monomer ratio, nitric oxide metabolites (nitrate plus nitrite), markers associated with endothelial activation, and vasoreactivity. Compared with controls, decompression stress increased NOX2 expression, reactive oxygen species and lipid peroxidation, decreased superoxide dismutase activity, reduced BH4 and nitric oxide metabolites. It also caused a shift in eNOS towards a lower dimer\/monomer ratio, increased endothelin-1 and adhesion molecules, and impaired endothelium-dependent relaxation, though endothelium-independent relaxation remained intact. GSK2795039 pretreatment attenuated oxidative stress, improved BH4 availability, restored nitric oxide metabolites, and decreased markers of endothelial activation, partially improving endothelium-dependent relaxation. These findings suggest that NOX2-associated oxidative stress contributes to reduced BH4 availability and eNOS coupling imbalance, leading to pulmonary endothelial dysfunction after decompression.<\/jats:p>","DOI":"10.1371\/journal.pone.0351145","type":"journal-article","created":{"date-parts":[[2026,7,2]],"date-time":"2026-07-02T17:43:09Z","timestamp":1783014189000},"page":"e0351145","update-policy":"https:\/\/doi.org\/10.1371\/journal.pone.corrections_policy","source":"Crossref","is-referenced-by-count":0,"title":["NOX2 inhibition attenuates oxidative stress and eNOS uncoupling in pulmonary arteries of rats following simulated air diving"],"prefix":"10.1371","volume":"21","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-9820-2409","authenticated-orcid":true,"given":"Qingbo","family":"Gong","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lin","family":"Xiao","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0009-0393-3782","authenticated-orcid":true,"given":"Song","family":"Wang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"340","published-online":{"date-parts":[[2026,7,2]]},"reference":[{"issue":"9760","key":"pone.0351145.ref001","doi-asserted-by":"crossref","first-page":"153","DOI":"10.1016\/S0140-6736(10)61085-9","article-title":"Decompression illness","volume":"377","author":"RD Vann","year":"2011","journal-title":"Lancet"},{"key":"pone.0351145.ref002","doi-asserted-by":"crossref","first-page":"901","DOI":"10.1113\/jphysiol.2005.089862","article-title":"A single air dive reduces arterial endothelial function in man","volume":"566","author":"AO Brubakk","year":"2005","journal-title":"J Physiol"},{"issue":"6","key":"pone.0351145.ref003","doi-asserted-by":"crossref","first-page":"432","DOI":"10.1111\/j.1475-097X.2010.00962.x","article-title":"Successive deep dives impair endothelial function and enhance oxidative stress in man","volume":"30","author":"A Obad","year":"2010","journal-title":"Clin Physiol Funct Imaging"},{"issue":"4","key":"pone.0351145.ref004","doi-asserted-by":"crossref","first-page":"389","DOI":"10.1016\/j.mehy.2008.11.022","article-title":"Gas bubbles may not be the underlying cause of decompression illness - The at-depth endothelial dysfunction hypothesis","volume":"72","author":"LA Madden","year":"2009","journal-title":"Med Hypotheses"},{"issue":"8","key":"pone.0351145.ref005","doi-asserted-by":"crossref","first-page":"1268","DOI":"10.1152\/japplphysiol.01305.2011","article-title":"Microparticle production, neutrophil activation, and intravascular bubbles following open-water SCUBA diving","volume":"112","author":"SR Thom","year":"2012","journal-title":"J Appl Physiol (1985)"},{"issue":"10","key":"pone.0351145.ref006","doi-asserted-by":"crossref","first-page":"1033","DOI":"10.1039\/c1ib00017a","article-title":"Mechanotransductional basis of endothelial cell response to intravascular bubbles","volume":"3","author":"AL Klinger","year":"2011","journal-title":"Integr Biol (Camb)"},{"issue":"6","key":"pone.0351145.ref007","doi-asserted-by":"crossref","first-page":"453","DOI":"10.1038\/nrd3403","article-title":"Combating oxidative stress in vascular disease: NADPH oxidases as therapeutic targets","volume":"10","author":"GR Drummond","year":"2011","journal-title":"Nat Rev Drug Discov"},{"issue":"9","key":"pone.0351145.ref008","doi-asserted-by":"crossref","first-page":"452","DOI":"10.1016\/j.tem.2014.06.012","article-title":"Endothelial NADPH oxidases: which NOX to target in vascular disease?","volume":"25","author":"GR Drummond","year":"2014","journal-title":"Trends Endocrinol Metab"},{"issue":"8","key":"pone.0351145.ref009","doi-asserted-by":"crossref","first-page":"1201","DOI":"10.1172\/JCI200314172","article-title":"Oxidation of tetrahydrobiopterin leads to uncoupling of endothelial cell nitric oxide synthase in hypertension","volume":"111","author":"U Landmesser","year":"2003","journal-title":"J Clin Invest"},{"issue":"13","key":"pone.0351145.ref010","doi-asserted-by":"crossref","first-page":"1708","DOI":"10.1161\/CIRCULATIONAHA.105.602532","article-title":"Endothelial nitric oxide synthase in vascular disease: from marvel to menace","volume":"113","author":"U F\u00f6rstermann","year":"2006","journal-title":"Circulation"},{"issue":"10","key":"pone.0351145.ref011","doi-asserted-by":"crossref","first-page":"840","DOI":"10.1161\/01.RES.87.10.840","article-title":"Endothelial dysfunction in cardiovascular diseases: the role of oxidant stress","volume":"87","author":"H Cai","year":"2000","journal-title":"Circ Res"},{"issue":"1","key":"pone.0351145.ref012","doi-asserted-by":"crossref","first-page":"87","DOI":"10.1111\/j.1742-7843.2011.00785.x","article-title":"Reactive oxygen species and endothelial function--role of nitric oxide synthase uncoupling and Nox family nicotinamide adenine dinucleotide phosphate oxidases","volume":"110","author":"AC Montezano","year":"2012","journal-title":"Basic Clin Pharmacol Toxicol"},{"issue":"3","key":"pone.0351145.ref013","doi-asserted-by":"crossref","first-page":"274","DOI":"10.1080\/10715760601080371","article-title":"Scuba diving enhances endogenous antioxidant defenses in lymphocytes and neutrophils","volume":"41","author":"MD Ferrer","year":"2007","journal-title":"Free Radic Res"},{"issue":"4","key":"pone.0351145.ref014","doi-asserted-by":"crossref","DOI":"10.1152\/ajpregu.00155.2015","article-title":"Ascorbic acid supplementation diminishes microparticle elevations and neutrophil activation following SCUBA diving","volume":"309","author":"M Yang","year":"2015","journal-title":"Am J Physiol Regul Integr Comp Physiol"},{"key":"pone.0351145.ref015","doi-asserted-by":"crossref","first-page":"239","DOI":"10.1016\/j.cis.2014.01.017","article-title":"Circulatory bubble dynamics: from physical to biological aspects","volume":"206","author":"V Papadopoulou","year":"2014","journal-title":"Adv Colloid Interface Sci"},{"key":"pone.0351145.ref016","doi-asserted-by":"crossref","first-page":"33390","DOI":"10.1038\/srep33390","article-title":"Endothelial dysfunction correlates with decompression bubbles in rats","volume":"6","author":"K Zhang","year":"2016","journal-title":"Sci Rep"},{"key":"pone.0351145.ref017","doi-asserted-by":"crossref","first-page":"611208","DOI":"10.3389\/fphys.2020.611208","article-title":"Extensive Simulated Diving Aggravates Endothelial Dysfunction in Male Pro-atherosclerotic ApoE Knockout Rats","volume":"11","author":"S Berenji Ardestani","year":"2020","journal-title":"Front Physiol"},{"issue":"8","key":"pone.0351145.ref018","article-title":"BM-MSCs mitigate lung injury in a rat model of decompression sickness","volume":"20","author":"C Lu","year":"2025","journal-title":"PLoS One"},{"key":"pone.0351145.ref019","doi-asserted-by":"crossref","first-page":"181","DOI":"10.3389\/fphys.2017.00181","article-title":"Time Course of Endothelial Dysfunction Induced by Decompression Bubbles in Rats","volume":"8","author":"K Zhang","year":"2017","journal-title":"Front Physiol"},{"key":"pone.0351145.ref020","first-page":"65","article-title":"Analysis of superoxide anion production in tissue","volume":"108","author":"M Zanetti","year":"2005","journal-title":"Methods Mol Med"},{"key":"pone.0351145.ref021","doi-asserted-by":"crossref","first-page":"100309","DOI":"10.1016\/j.conctc.2018.100309","article-title":"Comparing different ways of calculating sample size for two independent means: A worked example","volume":"13","author":"L Clifton","year":"2018","journal-title":"Contemp Clin Trials Commun"},{"key":"pone.0351145.ref022","doi-asserted-by":"crossref","first-page":"1","DOI":"10.28920\/dhm53.4.supp.1-53","article-title":"Decompression illness: a comprehensive overview","volume":"54","author":"SJ Mitchell","year":"2024","journal-title":"Diving Hyperb Med"},{"issue":"21","key":"pone.0351145.ref023","doi-asserted-by":"crossref","DOI":"10.3390\/ijms252111367","article-title":"Decompression illness after technical diving session in Mediterranean Sea: oxidative stress, inflammation, and HBO therapy","volume":"25","author":"S Mrakic-Sposta","year":"2024","journal-title":"Int J Mol Sci"},{"issue":"2","key":"pone.0351145.ref024","doi-asserted-by":"crossref","first-page":"435","DOI":"10.1152\/japplphysiol.00035.2021","article-title":"The protective effects of 1,3-butanediol acetoacetate diester on decompression sickness in rats","volume":"131","author":"K Zhang","year":"2021","journal-title":"J Appl Physiol (1985)"},{"issue":"17","key":"pone.0351145.ref025","doi-asserted-by":"crossref","first-page":"11632","DOI":"10.1021\/acs.jmedchem.3c00770","article-title":"NADPH oxidases: from molecular mechanisms to current inhibitors","volume":"66","author":"A Cipriano","year":"2023","journal-title":"J Med Chem"},{"key":"pone.0351145.ref026","doi-asserted-by":"crossref","first-page":"14","DOI":"10.1016\/j.freeradbiomed.2025.07.023","article-title":"Neutrophil extravasation and NETosis in decompression illness lung injury: A P-selectin\/PSGL-1\/NOX2\/PAD4 axis governed pathway validated by multi-model interrogation","volume":"239","author":"Z Li","year":"2025","journal-title":"Free Radic Biol Med"},{"issue":"5","key":"pone.0351145.ref027","doi-asserted-by":"crossref","first-page":"358","DOI":"10.1089\/ars.2014.6202","article-title":"Discovery of GSK2795039, a Novel Small Molecule NADPH Oxidase 2 Inhibitor","volume":"23","author":"K Hirano","year":"2015","journal-title":"Antioxid Redox Signal"},{"key":"pone.0351145.ref028","doi-asserted-by":"crossref","first-page":"114102","DOI":"10.1016\/j.jpba.2021.114102","article-title":"NOX2 inhibitor GSK2795039 metabolite identification towards drug optimization","volume":"201","author":"EC Padilha","year":"2021","journal-title":"J Pharm Biomed Anal"},{"issue":"1","key":"pone.0351145.ref029","doi-asserted-by":"crossref","first-page":"21","DOI":"10.1186\/s11658-023-00423-2","article-title":"Endothelial dysfunction due to eNOS uncoupling: molecular mechanisms as potential therapeutic targets","volume":"28","author":"A Janaszak-Jasiecka","year":"2023","journal-title":"Cell Mol Biol Lett"},{"issue":"2","key":"pone.0351145.ref030","doi-asserted-by":"crossref","first-page":"1136","DOI":"10.1074\/jbc.M805403200","article-title":"Quantitative regulation of intracellular endothelial nitric-oxide synthase (eNOS) coupling by both tetrahydrobiopterin-eNOS stoichiometry and biopterin redox status: insights from cells with tet-regulated GTP cyclohydrolase I expression","volume":"284","author":"MJ Crabtree","year":"2009","journal-title":"J Biol Chem"},{"issue":"4","key":"pone.0351145.ref031","doi-asserted-by":"crossref","first-page":"507","DOI":"10.1007\/s00421-008-0927-z","article-title":"Release of VCAM-1 associated endothelial microparticles following simulated SCUBA dives","volume":"105","author":"RV Vince","year":"2009","journal-title":"Eur J Appl Physiol"},{"issue":"13","key":"pone.0351145.ref032","doi-asserted-by":"crossref","first-page":"2278","DOI":"10.1093\/cvr\/cvad130","article-title":"Targeting endothelial vascular cell adhesion molecule-1 in atherosclerosis: drug discovery and development of vascular cell adhesion molecule-1-directed novel therapeutics","volume":"119","author":"JR Pickett","year":"2023","journal-title":"Cardiovasc Res"}],"container-title":["PLOS One"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/dx.plos.org\/10.1371\/journal.pone.0351145","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,7,2]],"date-time":"2026-07-02T17:43:16Z","timestamp":1783014196000},"score":1,"resource":{"primary":{"URL":"https:\/\/dx.plos.org\/10.1371\/journal.pone.0351145"}},"subtitle":[],"editor":[{"given":"Peng","family":"Zhang","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"editor"}]}],"short-title":[],"issued":{"date-parts":[[2026,7,2]]},"references-count":32,"journal-issue":{"issue":"7","published-online":{"date-parts":[[2026,7,2]]}},"URL":"https:\/\/doi.org\/10.1371\/journal.pone.0351145","relation":{},"ISSN":["1932-6203"],"issn-type":[{"value":"1932-6203","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,7,2]]}}}