{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,24]],"date-time":"2026-07-24T14:30:17Z","timestamp":1784903417348,"version":"3.55.0"},"reference-count":39,"publisher":"Public Library of Science (PLoS)","issue":"5","license":[{"start":{"date-parts":[[2024,5,15]],"date-time":"2024-05-15T00:00:00Z","timestamp":1715731200000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"NIH\/NEI","award":["EY019949, EY025061, EY030970"],"award-info":[{"award-number":["EY019949, EY025061, EY030970"]}]},{"DOI":"10.13039\/100006312","name":"BrightFocus Foundation","doi-asserted-by":"publisher","award":["NGR G2019302"],"award-info":[{"award-number":["NGR G2019302"]}],"id":[{"id":"10.13039\/100006312","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100001818","name":"Research to Prevent Blindness","doi-asserted-by":"publisher","award":["An Unrestricted Grant to the Department of Ophthalmology, the State University of New York at Buffalo"],"award-info":[{"award-number":["An Unrestricted Grant to the Department of Ophthalmology, the State University of New York at Buffalo"]}],"id":[{"id":"10.13039\/100001818","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["www.plosone.org"],"crossmark-restriction":false},"short-container-title":["PLoS ONE"],"abstract":"<jats:p>Diabetic Retinopathy (DR) is the leading cause of vision loss in working-age adults. The hallmark features of DR include vascular leakage, capillary loss, retinal ischemia, and aberrant neovascularization. Although the pathophysiology is not fully understood, accumulating evidence supports elevated reactive oxygen species associated with increased activity of NADPH oxidase 4 (Nox4) as major drivers of disease progression. Previously, we have shown that Nox4 upregulation in retinal endothelial cells by diabetes leads to increased vascular leakage by an unknown mechanism. Platelet endothelial cell adhesion molecule 1 (PECAM-1) is a cell surface molecule that is highly expressed in endothelial cells and regulates endothelial barrier function. In the present study, using endothelial cell-specific human Nox4 transgenic (TG) mice and endothelial cell-specific Nox4 conditional knockout (cKO) mice, we investigated the impact of Nox4 upregulation on PECAM-1 expression in mouse retinas and brain microvascular endothelial cells (BMECs). Additionally, cultured human retinal endothelial cells (HRECs) transduced with adenovirus overexpressing human Nox4 were used in the study. We found that overexpression of Nox4 increases PECAM-1 mRNA but has no effect on its protein expression in the mouse retina, BMECs, or HRECs. Furthermore, PECAM-1 mRNA and protein expression was unchanged in BMECs isolated from cKO mice compared to wild type (WT) mice with or without 2 months of diabetes. Together, these findings do not support a significant role of Nox4 in the regulation of PECAM-1 expression in the diabetic retina and endothelial cells. Further studies are warranted to elucidate the mechanism of Nox4-induced vascular leakage by investigating other intercellular junctional proteins in endothelial cells and their implications in the pathophysiology of diabetic retinopathy.<\/jats:p>","DOI":"10.1371\/journal.pone.0303010","type":"journal-article","created":{"date-parts":[[2024,5,15]],"date-time":"2024-05-15T17:38:57Z","timestamp":1715794737000},"page":"e0303010","update-policy":"https:\/\/doi.org\/10.1371\/journal.pone.corrections_policy","source":"Crossref","is-referenced-by-count":2,"title":["Effects of Nox4 upregulation on PECAM-1 expression in a mouse model of diabetic retinopathy"],"prefix":"10.1371","volume":"19","author":[{"given":"Jinli","family":"Wang","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0008-4796-7661","authenticated-orcid":true,"given":"Daniel A.","family":"Lai","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Joshua J.","family":"Wang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7848-3263","authenticated-orcid":true,"given":"Sarah X.","family":"Zhang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"340","published-online":{"date-parts":[[2024,5,15]]},"reference":[{"issue":"9735","key":"pone.0303010.ref001","doi-asserted-by":"crossref","first-page":"124","DOI":"10.1016\/S0140-6736(09)62124-3","article-title":"Diabetic retinopathy.","volume":"376","author":"N Cheung","year":"2010","journal-title":"Lancet"},{"issue":"6","key":"pone.0303010.ref002","doi-asserted-by":"crossref","DOI":"10.3390\/ijms19061816","article-title":"Diabetic Retinopathy: Pathophysiology and Treatments.","volume":"19","author":"W Wang","year":"2018","journal-title":"Int J Mol Sci."},{"issue":"6","key":"pone.0303010.ref003","doi-asserted-by":"crossref","first-page":"1528","DOI":"10.2337\/db09-1057","article-title":"Inhibition of reactive oxygen species by Lovastatin downregulates vascular endothelial growth factor expression and ameliorates blood-retinal barrier breakdown in db\/db mice: role of NADPH oxidase 4","volume":"59","author":"J Li","year":"2010","journal-title":"Diabetes"},{"issue":"2","key":"pone.0303010.ref004","doi-asserted-by":"crossref","first-page":"748","DOI":"10.1167\/iovs.18-25068","article-title":"Loss of Platelet Endothelial Cell Adhesion Molecule-1 (PECAM-1) in the Diabetic Retina: Role of Matrix Metalloproteinases.","volume":"60","author":"RS Eshaq","year":"2019","journal-title":"Invest Ophthalmol Vis Sci"},{"key":"pone.0303010.ref005","doi-asserted-by":"crossref","first-page":"101799","DOI":"10.1016\/j.redox.2020.101799","article-title":"Oxidative stress and diabetic retinopathy: Molecular mechanisms, pathogenetic role and therapeutic implications.","volume":"37","author":"Q Kang","year":"2020","journal-title":"Redox Biol.PubMed Central PMCID"},{"issue":"4","key":"pone.0303010.ref006","doi-asserted-by":"crossref","first-page":"C719","DOI":"10.1152\/ajpcell.2001.280.4.C719","article-title":"Oxidant stress and endothelial cell dysfunction","volume":"280","author":"H Lum","year":"2001","journal-title":"Am J Physiol Cell Physiol"},{"issue":"4","key":"pone.0303010.ref007","doi-asserted-by":"crossref","first-page":"R1619","DOI":"10.1152\/ajpregu.00290.2007","article-title":"Role of NADPH oxidase and ANG II in diabetes-induced retinal leukostasis","volume":"293","author":"P Chen","year":"2007","journal-title":"Am J Physiol Regul Integr Comp Physiol"},{"issue":"12","key":"pone.0303010.ref008","doi-asserted-by":"crossref","first-page":"6167","DOI":"10.1158\/0008-5472.CAN-05-3640","article-title":"Hydrogen peroxide produced by angiopoietin-1 mediates angiogenesis","volume":"66","author":"YM Kim","year":"2006","journal-title":"Cancer research"},{"issue":"5","key":"pone.0303010.ref009","article-title":"Diabetic Retinopathy and NADPH Oxidase-2: A Sweet Slippery Road.","volume":"10","author":"RA Kowluru","year":"2021","journal-title":"Antioxidants (Basel, Switzerland)."},{"key":"pone.0303010.ref010","first-page":"231","article-title":"Endothelial NADPH oxidase 4 mediates vascular endothelial growth factor receptor 2-induced intravitreal neovascularization in a rat model of retinopathy of prematurity","volume":"20","author":"H Wang","year":"2014","journal-title":"Mol Vis"},{"issue":"1","key":"pone.0303010.ref011","doi-asserted-by":"crossref","first-page":"112","DOI":"10.2337\/db22-0194","article-title":"Sustained Upregulation of Endothelial Nox4 Mediates Retinal Vascular Pathology in Type 1","volume":"72","author":"X Tang","year":"2023","journal-title":"Diabetes. 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