{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,26]],"date-time":"2026-08-26T10:25:40Z","timestamp":1787739940204,"version":"build-2784847793"},"reference-count":43,"publisher":"Public Library of Science (PLoS)","issue":"6","license":[{"start":{"date-parts":[[2025,6,20]],"date-time":"2025-06-20T00:00:00Z","timestamp":1750377600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Medical Reserve Talent Cultivation Program of Yunnan Provincial Health Commission","award":["H-2018021"],"award-info":[{"award-number":["H-2018021"]}]},{"name":"Medical Leading Talent Training Program of Yunnan Provincial Health Commission","award":["L-2019029"],"award-info":[{"award-number":["L-2019029"]}]}],"content-domain":{"domain":["www.plosone.org"],"crossmark-restriction":false},"short-container-title":["PLoS One"],"abstract":"\nBackground<\/jats:title>\nChoroidal neovascularization (CNV) is a key manifestation of intraocular neovascularization, and it is considered one of the main causes of blindness in ophthalmology. Additionally, multiple anti-vascular endothelial growth factor (VEGF) drugs have been used as first-line treatment for CNV. However, several issues posed challenges to the anti-VEGF drugs, which were mainly composed of short duration of action, requirement for repeated injections, and complications. Thrombospondin-1 (TSP-1) is an endogenous protein that was found to regulate multiple biological processes within the body, and it has been proven to exhibit an inhibitory effect on neovascularization. Besides, the function of TSP-1 during the inhibition of neovascularization was currently considered to mainly focus on its type \u2160 repeats (TSRs), which was attributed to the large molecular weight, complex structure, and possible unknown functions of TSP-1. Therefore, TSRs can be applied as targets and research directions for the further development and exploration of potential therapeutic drugs.<\/jats:p>\n<\/jats:sec>\n\nObjectives<\/jats:title>\nBased on the type I repeats (TSRs) of thrombospondin-1 (TSP-1), amino acid sequences of different lengths were designed and synthesized in this study, named as VR-9 VR-10\u3001VR-11\u3001VR-12\u3001VR-13. The objective was to explore the effects of the above five peptides on angiogenesis in Chori-retinal neovascularization, alongside the screening of the best peptides and the deep exploration into the underlying mechanism, aimed to provide a basis for the development and application of peptide drugs in the treatment of CNV.<\/jats:p>\n<\/jats:sec>\n\nMethods<\/jats:title>\nWound healing, CCK-8, and 5-ethynyl-2\u2032-deoxyuridine (EdU) assays were employed to evaluate the proliferation and migration ability of cells. CRISPR-Cas9 technology was utilized to establish CD36 knockdown cell lines, alongside the conduction of qPCR to verify the efficiency of gene knockdown. The expression levels of VEGF and CD31 in RF\/6A cells and rats were assessed by Western blot. Additionally, Hematoxylin and eosin (HE) staining was performed to examine the structural integrity of the rat retina, while Fluorescein Isothiocyanate-Dextran Cardiac Perfusion (FITC) labeling was used to observe the occurrence and development of choroidal neovascularization (CNV).<\/jats:p>\n<\/jats:sec>\n\nResults<\/jats:title>\nAccording to the wound-healing and CCK-8 assays, VR-13 was the most effective in inhibiting the proliferation and migration of endothelial cells. Furthermore, VR-13 peptide effectively inhibited the pathological development of CNV without the detection of retinal toxicity in the rat CNV model.<\/jats:p>\n<\/jats:sec>\n\nConclusions<\/jats:title>\nOverall, it was found that VR-13 exhibit significant effects on the inducing of apoptosis and the inhibition of the progression of angiogenesis by regulating the expression of VEGF and CD31 via CD36 signaling pathway.<\/jats:p>\n<\/jats:sec>","DOI":"10.1371\/journal.pone.0325661","type":"journal-article","created":{"date-parts":[[2025,6,20]],"date-time":"2025-06-20T17:24:48Z","timestamp":1750440288000},"page":"e0325661","update-policy":"https:\/\/doi.org\/10.1371\/journal.pone.corrections_policy","source":"Crossref","is-referenced-by-count":2,"title":["Active peptides of TSP-1 inhibit retinal angiogenesis through the CD36 pathway in a rat model of choroidal neovascularization"],"prefix":"10.1371","volume":"20","author":[{"given":"Yadi","family":"Li","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Aiping","family":"Deng","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kangwei","family":"Jiao","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jie","family":"Yan","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wandong","family":"Zuo","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yujie","family":"Dong","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wenrong","family":"Xu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yuting","family":"Li","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chunming","family":"Guo","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Maorong","family":"Chen","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Run","family":"Tian","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1575-5356","authenticated-orcid":true,"given":"Zhulin","family":"Hu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"340","published-online":{"date-parts":[[2025,6,20]]},"reference":[{"key":"pone.0325661.ref001","doi-asserted-by":"crossref","first-page":"101113","DOI":"10.1016\/j.preteyeres.2022.101113","article-title":"Towards a better understanding of non-exudative choroidal and macular neovascularization","volume":"92","author":"R Sacconi","year":"2023","journal-title":"Prog Retin Eye Res"},{"issue":"6","key":"pone.0325661.ref002","article-title":"Anti-vascular endothelial growth factor biosimilars for neovascular age-related macular degeneration","volume":"6","author":"T Sunaga","year":"2024","journal-title":"Cochrane Database Syst Rev"},{"issue":"2","key":"pone.0325661.ref003","doi-asserted-by":"crossref","first-page":"600","DOI":"10.1177\/1120672120902028","article-title":"Incidence of endophthalmitis after intravitreal injection with and without topical antibiotic prophylaxis","volume":"31","author":"S Torres-Costa","year":"2021","journal-title":"Eur J Ophthalmol"},{"issue":"21","key":"pone.0325661.ref004","doi-asserted-by":"crossref","first-page":"8242","DOI":"10.3390\/ijms21218242","article-title":"Neovascular age-related macular degeneration: therapeutic management and new-upcoming approaches","volume":"21","author":"F Ricci","year":"2020","journal-title":"Int J Mol Sci"},{"issue":"5","key":"pone.0325661.ref005","doi-asserted-by":"crossref","first-page":"28","DOI":"10.1167\/iovs.64.5.28","article-title":"Mechanisms of acquired resistance to anti-VEGF therapy for neovascular eye diseases","volume":"64","author":"D Sharma","year":"2023","journal-title":"Invest Ophthalmol Vis Sci"},{"key":"pone.0325661.ref006","doi-asserted-by":"crossref","first-page":"100906","DOI":"10.1016\/j.preteyeres.2020.100906","article-title":"Incomplete response to Anti-VEGF therapy in neovascular AMD: Exploring disease mechanisms and therapeutic opportunities","volume":"82","author":"PS Mettu","year":"2021","journal-title":"Prog Retin Eye Res"},{"issue":"10","key":"pone.0325661.ref007","doi-asserted-by":"crossref","first-page":"982","DOI":"10.1002\/iub.2610","article-title":"Thrombospondins in human aortic aneurysms","volume":"74","author":"E Plana","year":"2022","journal-title":"IUBMB Life"},{"issue":"7","key":"pone.0325661.ref008","doi-asserted-by":"crossref","first-page":"376","DOI":"10.1089\/jop.2015.0020","article-title":"Thrombospondin-1 as a regulator of corneal inflammation and lymphangiogenesis: effects on dry eye disease and corneal graft immunology","volume":"31","author":"L Sch\u00f6llhorn","year":"2015","journal-title":"J Ocul Pharmacol Ther"},{"key":"pone.0325661.ref009","doi-asserted-by":"crossref","first-page":"22","DOI":"10.1016\/j.semcdb.2023.05.008","article-title":"Emerging functions of thrombospondin-1 in immunity","volume":"155","author":"S Kaur","year":"2024","journal-title":"Semin Cell Dev Biol"},{"issue":"4","key":"pone.0325661.ref010","doi-asserted-by":"crossref","DOI":"10.15190\/d.2014.23","article-title":"Thrombospondin-1 modulates actin filament remodeling and cell motility in mouse mammary tumor cells in vitro","volume":"2","author":"D Ndishabandi","year":"2014","journal-title":"Discoveries (Craiova)"},{"issue":"41","key":"pone.0325661.ref011","doi-asserted-by":"crossref","first-page":"42393","DOI":"10.1074\/jbc.M407631200","article-title":"Mechanism of CD47-induced alpha4beta1 integrin activation and adhesion in sickle reticulocytes","volume":"279","author":"JE Brittain","year":"2004","journal-title":"J Biol Chem"},{"issue":"2","key":"pone.0325661.ref012","doi-asserted-by":"crossref","first-page":"609","DOI":"10.1159\/000488628","article-title":"VR-10 thrombospondin-1 synthetic polypeptide\u2019s impact on rhesus choroid-retinal endothelial cells","volume":"46","author":"R Tian","year":"2018","journal-title":"Cell Physiol Biochem"},{"issue":"10","key":"pone.0325661.ref013","doi-asserted-by":"crossref","first-page":"1345","DOI":"10.1016\/j.ophtha.2020.04.017","article-title":"Retinal vasculitis and intraocular inflammation after intravitreal injection of brolucizumab","volume":"127","author":"CR Baumal","year":"2020","journal-title":"Ophthalmology"},{"key":"pone.0325661.ref014","doi-asserted-by":"crossref","first-page":"100680","DOI":"10.1016\/j.ajoc.2020.100680","article-title":"Retinal arterial occlusive vasculitis following intravitreal brolucizumab administration","volume":"18","author":"SJ Haug","year":"2020","journal-title":"Am J Ophthalmol Case Rep"},{"issue":"7","key":"pone.0325661.ref015","doi-asserted-by":"crossref","first-page":"1050","DOI":"10.1016\/j.ophtha.2020.11.011","article-title":"Risk of inflammation, retinal vasculitis, and retinal occlusion-related events with brolucizumab: post hoc review of HAWK and HARRIER","volume":"128","author":"J Mon\u00e9s","year":"2021","journal-title":"Ophthalmology"},{"issue":"5","key":"pone.0325661.ref016","first-page":"1275","article-title":"Role of thrombospondin\u20111 and thrombospondin\u20112 in cardiovascular diseases (Review)","volume":"45","author":"K Zhang","year":"2020","journal-title":"Int J Mol Med"},{"key":"pone.0325661.ref017","doi-asserted-by":"crossref","first-page":"45","DOI":"10.1016\/j.semcdb.2023.06.009","article-title":"Thrombospondin-1 in drug activity and tumor response to therapies","volume":"155","author":"E Longhi","year":"2024","journal-title":"Semin Cell Dev Biol"},{"issue":"9","key":"pone.0325661.ref018","doi-asserted-by":"crossref","first-page":"4570","DOI":"10.3390\/ijms22094570","article-title":"Functions of thrombospondin-1 in the tumor microenvironment","volume":"22","author":"S Kaur","year":"2021","journal-title":"Int J Mol Sci"},{"issue":"9","key":"pone.0325661.ref019","doi-asserted-by":"crossref","first-page":"2335","DOI":"10.1158\/1078-0432.CCR-16-1520","article-title":"Dysregulation of Rab37-mediated cross-talk between cancer cells and endothelial cells via thrombospondin-1 promotes tumor neovasculature and metastasis","volume":"23","author":"H-T Tzeng","year":"2017","journal-title":"Clin Cancer Res"},{"issue":"3","key":"pone.0325661.ref020","doi-asserted-by":"crossref","first-page":"673","DOI":"10.3390\/ijms20030673","article-title":"Diabetes impairs angiogenesis and induces endothelial cell senescence by up-regulating thrombospondin-CD47-dependent signaling","volume":"20","author":"MS Bitar","year":"2019","journal-title":"Int J Mol Sci"},{"issue":"1","key":"pone.0325661.ref021","doi-asserted-by":"crossref","first-page":"42","DOI":"10.1016\/j.cjca.2018.11.008","article-title":"Thrombospondin 1 is increased in the aorta and plasma of patients with acute aortic dissection","volume":"35","author":"T Zeng","year":"2019","journal-title":"Can J Cardiol"},{"issue":"2","key":"pone.0325661.ref022","doi-asserted-by":"crossref","first-page":"81","DOI":"10.1159\/000362371","article-title":"Thrombospondin-1 is produced by retinal glial cells and inhibits the growth of vascular endothelial cells","volume":"52","author":"Y Yafai","year":"2014","journal-title":"Ophthalmic Res"},{"issue":"8","key":"pone.0325661.ref023","doi-asserted-by":"crossref","first-page":"1472","DOI":"10.1161\/01.ATV.0000081636.25235.C6","article-title":"Novel proangiogenic effect of factor XIII associated with suppression of thrombospondin 1 expression","volume":"23","author":"R Dardik","year":"2003","journal-title":"Arterioscler Thromb Vasc Biol"},{"issue":"22","key":"pone.0325661.ref024","doi-asserted-by":"crossref","first-page":"5188","DOI":"10.1200\/JCO.2005.05.013","article-title":"Phase I safety, pharmacokinetic, and pharmacodynamic study of the thrombospondin-1-mimetic angiogenesis inhibitor ABT-510 in patients with advanced cancer","volume":"23","author":"R Hoekstra","year":"2005","journal-title":"J Clin Oncol"},{"issue":"24","key":"pone.0325661.ref025","doi-asserted-by":"crossref","first-page":"7444","DOI":"10.1158\/1078-0432.CCR-06-0109","article-title":"Preclinical evaluation of antiangiogenic thrombospondin-1 peptide mimetics, ABT-526 and ABT-510, in companion dogs with naturally occurring cancers","volume":"12","author":"A Rusk","year":"2006","journal-title":"Clin Cancer Res"},{"issue":"6","key":"pone.0325661.ref026","doi-asserted-by":"crossref","first-page":"685","DOI":"10.1164\/rccm.202104-1021OC","article-title":"Thrombospondin-1 plays a major pathogenic role in experimental and human bronchopulmonary dysplasia","volume":"205","author":"BA Ruschkowski","year":"2022","journal-title":"Am J Respir Crit Care Med"},{"issue":"2","key":"pone.0325661.ref027","doi-asserted-by":"crossref","first-page":"310","DOI":"10.1161\/CIRCRESAHA.120.316851","article-title":"Basic and therapeutic aspects of angiogenesis updated","volume":"127","author":"G Eelen","year":"2020","journal-title":"Circ Res"},{"key":"pone.0325661.ref028","doi-asserted-by":"crossref","first-page":"126","DOI":"10.1016\/j.semcancer.2022.09.006","article-title":"Counter regulation of tumor angiogenesis by vascular endothelial growth factor and thrombospondin-1","volume":"86","author":"J Lawler","year":"2022","journal-title":"Semin Cancer Biol"},{"issue":"3","key":"pone.0325661.ref029","doi-asserted-by":"crossref","first-page":"375","DOI":"10.1007\/s10456-024-09919-7","article-title":"Correction: Granzyme B degrades extracellular matrix and promotes inflammation and choroidal neovascularization","volume":"27","author":"G Obasanmi","year":"2024","journal-title":"Angiogenesis"},{"issue":"11","key":"pone.0325661.ref030","doi-asserted-by":"crossref","first-page":"2768","DOI":"10.1093\/emboj\/20.11.2768","article-title":"A single autophosphorylation site on KDR\/Flk-1 is essential for VEGF-A-dependent activation of PLC-gamma and DNA synthesis in vascular endothelial cells","volume":"20","author":"T Takahashi","year":"2001","journal-title":"EMBO J"},{"issue":"6","key":"pone.0325661.ref031","doi-asserted-by":"crossref","first-page":"1248","DOI":"10.1016\/j.cell.2019.01.021","article-title":"VEGF in signaling and disease: beyond discovery and development","volume":"176","author":"RS Apte","year":"2019","journal-title":"Cell"},{"issue":"11","key":"pone.0325661.ref032","doi-asserted-by":"crossref","first-page":"816","DOI":"10.1038\/s41580-023-00631-w","article-title":"Biology and therapeutic targeting of vascular endothelial growth factor A","volume":"24","author":"L P\u00e9rez-Guti\u00e9rrez","year":"2023","journal-title":"Nat Rev Mol Cell Biol"},{"issue":"4","key":"pone.0325661.ref033","doi-asserted-by":"crossref","first-page":"691","DOI":"10.1016\/j.cell.2007.06.054","article-title":"Autocrine VEGF signaling is required for vascular homeostasis","volume":"130","author":"S Lee","year":"2007","journal-title":"Cell"},{"issue":"7","key":"pone.0325661.ref034","doi-asserted-by":"crossref","first-page":"1341","DOI":"10.1242\/jcs.159954","article-title":"Local VE-cadherin mechanotransduction triggers long-ranged remodeling of endothelial monolayers","volume":"128","author":"AK Barry","year":"2015","journal-title":"J Cell Sci"},{"issue":"7","key":"pone.0325661.ref035","doi-asserted-by":"crossref","first-page":"1329","DOI":"10.1091\/mbc.8.7.1329","article-title":"Thrombospondin-1, a natural inhibitor of angiogenesis, regulates platelet-endothelial cell adhesion molecule-1 expression and endothelial cell morphogenesis","volume":"8","author":"N Sheibani","year":"1997","journal-title":"Mol Biol Cell"},{"issue":"3","key":"pone.0325661.ref036","doi-asserted-by":"crossref","first-page":"887","DOI":"10.1016\/S0002-9440(10)65188-7","article-title":"Functional roles for PECAM-1 (CD31) and VE-cadherin (CD144) in tube assembly and lumen formation in three-dimensional collagen gels","volume":"155","author":"S Yang","year":"1999","journal-title":"Am J Pathol"},{"issue":"5","key":"pone.0325661.ref037","article-title":"Involvement of human PECAM-1 in angiogenesis and in vitro endothelial cell migration","volume":"282","author":"G Cao","year":"2002","journal-title":"Am J Physiol Cell Physiol"},{"issue":"2","key":"pone.0325661.ref038","doi-asserted-by":"crossref","first-page":"903","DOI":"10.2353\/ajpath.2009.090206","article-title":"Angiogenesis in platelet endothelial cell adhesion molecule-1-null mice","volume":"175","author":"G Cao","year":"2009","journal-title":"Am J Pathol"},{"issue":"3","key":"pone.0325661.ref039","doi-asserted-by":"crossref","first-page":"253","DOI":"10.1097\/MOH.0000000000000239","article-title":"Endothelial functions of platelet\/endothelial cell adhesion molecule-1 (CD31)","volume":"23","author":"P Lertkiatmongkol","year":"2016","journal-title":"Curr Opin Hematol"},{"issue":"9","key":"pone.0325661.ref040","doi-asserted-by":"crossref","first-page":"1178","DOI":"10.1161\/CIRCRESAHA.120.315935","article-title":"CD31 as a therapeutic target in atherosclerosis","volume":"126","author":"G Caligiuri","year":"2020","journal-title":"Circ Res"},{"issue":"4","key":"pone.0325661.ref041","doi-asserted-by":"crossref","first-page":"308","DOI":"10.1089\/ars.2013.5743","article-title":"Oxidized high-density lipoprotein impairs endothelial progenitor cells\u2019 function by activation of CD36-MAPK-TSP-1 pathways","volume":"22","author":"J Wu","year":"2015","journal-title":"Antioxid Redox Signal"},{"issue":"1","key":"pone.0325661.ref042","doi-asserted-by":"crossref","first-page":"171","DOI":"10.3390\/cells12010171","article-title":"The past and present lives of the intraocular transmembrane protein CD36","volume":"12","author":"R Yang","year":"2022","journal-title":"Cells"},{"key":"pone.0325661.ref043","doi-asserted-by":"crossref","first-page":"170868","DOI":"10.1016\/j.peptides.2022.170868","article-title":"VR-10 polypeptide interacts with CD36 to induce cell apoptosis and autophagy in choroid-retinal endothelial cells: Identification of VR-10 as putative novel therapeutic agent for choroid neovascularization (CNV) treatment","volume":"157","author":"R Tian","year":"2022","journal-title":"Peptides"}],"container-title":["PLOS One"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/dx.plos.org\/10.1371\/journal.pone.0325661","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,6,20]],"date-time":"2025-06-20T17:24:57Z","timestamp":1750440297000},"score":1,"resource":{"primary":{"URL":"https:\/\/dx.plos.org\/10.1371\/journal.pone.0325661"}},"subtitle":[],"editor":[{"given":"Keun-Yeong","family":"Jeong","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"editor"}]}],"short-title":[],"issued":{"date-parts":[[2025,6,20]]},"references-count":43,"journal-issue":{"issue":"6","published-online":{"date-parts":[[2025,6,20]]}},"URL":"https:\/\/doi.org\/10.1371\/journal.pone.0325661","relation":{},"ISSN":["1932-6203"],"issn-type":[{"value":"1932-6203","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,6,20]]}}}