{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,2,19]],"date-time":"2026-02-19T03:04:23Z","timestamp":1771470263814,"version":"3.50.1"},"reference-count":45,"publisher":"Portland Press Ltd.","issue":"3","license":[{"start":{"date-parts":[[2021,1,18]],"date-time":"2021-01-18T00:00:00Z","timestamp":1610928000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/2.0\/"}],"content-domain":{"domain":["portlandpress.com"],"crossmark-restriction":true},"short-container-title":[],"published-print":{"date-parts":[[2021,2,12]]},"abstract":"<jats:title>Abstract<\/jats:title>\n               <jats:p>Abnormal vascular smooth muscle cell (VSMC) proliferation is a critical step in the development of atherosclerosis. Serpina3c is a serine protease inhibitor (serpin) that plays a key role in metabolic diseases. The present study aimed to investigate the role of serpina3c in atherosclerosis and regulation of VSMC proliferation and possible mechanisms. Serpina3c is down-regulated during high-fat diet (HFD)-induced atherosclerosis. An Apoe\u2212\/\u2212\/serpina3c\u2212\/\u2212-double-knockout mouse model was used to determine the role of serpina3c in atherosclerosis after HFD for 12 weeks. Compared with Apoe\u2212\/\u2212 mice, the Apoe\u2212\/\u2212\/serpina3c\u2212\/\u2212 mice developed more severe atherosclerosis, and the number of VSMCs and macrophages in aortic plaques was significantly increased. The present study revealed serpina3c as a novel thrombin inhibitor that suppressed thrombin activity. In circulating plasma, thrombin activity was high in the Apoe\u2212\/\u2212\/serpina3c\u2212\/\u2212 mice, compared with Apoe\u2212\/\u2212 mice. Immunofluorescence staining showed thrombin and serpina3c colocalization in the liver and aortic cusp. In addition, inhibition of thrombin by dabigatran in serpina3c\u2212\/\u2212 mice reduced neointima lesion formation due to partial carotid artery ligation. Moreover, an in vitro study confirmed that thrombin activity was also decreased by serpina3c protein, supernatant and cell lysate that overexpressed serpina3c. The results of experiments showed that serpina3c negatively regulated VSMC proliferation in culture. The possible mechanism may involve serpina3c inhibition of ERK1\/2 and JNK signaling in thrombin\/PAR-1 system-mediated VSMC proliferation. Our results highlight a protective role for serpina3c as a novel thrombin inhibitor in the development of atherosclerosis, with serpina3c conferring protection through the thrombin\/PAR-1 system to negatively regulate VSMC proliferation through ERK1\/2 and JNK signaling.<\/jats:p>","DOI":"10.1042\/cs20201235","type":"journal-article","created":{"date-parts":[[2021,1,18]],"date-time":"2021-01-18T09:51:53Z","timestamp":1610963513000},"page":"447-463","update-policy":"https:\/\/doi.org\/10.1042\/crossmark_policy","source":"Crossref","is-referenced-by-count":14,"title":["Protective role of serpina3c as a novel thrombin inhibitor against atherosclerosis in mice"],"prefix":"10.1042","volume":"135","author":[{"given":"Ling-lin","family":"Qian","sequence":"first","affiliation":[{"name":"Department of Cardiology, Zhongda Hospital, School of Medicine, Southeast University, 87 Dingjiaqiao, Nanjing, Jiangsu 210009, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jing-jing","family":"Ji","sequence":"additional","affiliation":[{"name":"Department of Cardiology, Zhongda Hospital, School of Medicine, Southeast University, 87 Dingjiaqiao, Nanjing, Jiangsu 210009, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jia-qi","family":"Guo","sequence":"additional","affiliation":[{"name":"Department of Cardiology, Zhongda Hospital, School of Medicine, Southeast University, 87 Dingjiaqiao, Nanjing, Jiangsu 210009, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yan-ping","family":"Wu","sequence":"additional","affiliation":[{"name":"Department of Cardiology, Zhongda Hospital, School of Medicine, Southeast University, 87 Dingjiaqiao, Nanjing, Jiangsu 210009, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Gen-shan","family":"Ma","sequence":"additional","affiliation":[{"name":"Department of Cardiology, Zhongda Hospital, School of Medicine, Southeast University, 87 Dingjiaqiao, Nanjing, Jiangsu 210009, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7841-288X","authenticated-orcid":false,"given":"Yu-yu","family":"Yao","sequence":"additional","affiliation":[{"name":"Department of Cardiology, Zhongda Hospital, School of Medicine, Southeast University, 87 Dingjiaqiao, Nanjing, Jiangsu 210009, China"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"288","published-online":{"date-parts":[[2021,2,1]]},"reference":[{"key":"2021113011191650600_B1","doi-asserted-by":"publisher","first-page":"531","DOI":"10.1161\/CIRCRESAHA.116.308334","article-title":"Atherosclerosis: Successes, Surprises, and Future Challenges","volume":"118","author":"Libby","year":"2016","journal-title":"Circ. Res."},{"key":"2021113011191650600_B2","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1160\/TH10-12-0784","article-title":"Stabilisation of atherosclerotic plaques. Position paper of the European Society of Cardiology (ESC) Working Group on atherosclerosis and vascular biology","volume":"106","author":"Yl\u00e4-Herttuala","year":"2011","journal-title":"Thromb. Haemost."},{"key":"2021113011191650600_B3","doi-asserted-by":"publisher","first-page":"104","DOI":"10.1161\/CIRCINTERVENTIONS.110.957332","article-title":"Vascular smooth muscle cell proliferation in restenosis","volume":"4","author":"Marx","year":"2011","journal-title":"Circulation. Cardiovasc. Interven."},{"key":"2021113011191650600_B4","doi-asserted-by":"publisher","first-page":"812","DOI":"10.1161\/ATVBAHA.107.159327","article-title":"Role of smooth muscle cells in the initiation and early progression of atherosclerosis","volume":"28","author":"Doran","year":"2008","journal-title":"Arterioscler. Thromb. Vasc. Biol."},{"key":"2021113011191650600_B5","doi-asserted-by":"publisher","first-page":"767","DOI":"10.1152\/physrev.00041.2003","article-title":"Molecular regulation of vascular smooth muscle cell differentiation in development and disease","volume":"84","author":"Owens","year":"2004","journal-title":"Physiol. Rev."},{"key":"2021113011191650600_B6","doi-asserted-by":"publisher","first-page":"15957","DOI":"10.1016\/S0021-9258(17)33954-6","article-title":"The serpin superfamily of proteinase inhibitors: structure, function, and regulation","volume":"269","author":"Potempa","year":"1994","journal-title":"J. Biol. Chem."},{"key":"2021113011191650600_B7","doi-asserted-by":"publisher","first-page":"5258","DOI":"10.1021\/bi972521d","article-title":"Cross-class inhibition of the cysteine proteinases cathepsins K, L, and S by the serpin squamous cell carcinoma antigen 1: a kinetic analysis","volume":"37","author":"Schick","year":"1998","journal-title":"Biochemistry"},{"key":"2021113011191650600_B8","doi-asserted-by":"publisher","first-page":"73","DOI":"10.1038\/s41420-019-0152-z","article-title":"The whole transcriptome and proteome changes in the early stage of myocardial infarction","volume":"5","author":"Li","year":"2019","journal-title":"Cell Death Discovery"},{"key":"2021113011191650600_B9","doi-asserted-by":"publisher","first-page":"288","DOI":"10.3892\/ijmm.2012.994","article-title":"Serine protease inhibitor A3 in atherosclerosis and aneurysm disease","volume":"30","author":"W\u00e5gs\u00e4ter","year":"2012","journal-title":"Int. J. Mol. Med."},{"key":"2021113011191650600_B10","doi-asserted-by":"publisher","first-page":"368","DOI":"10.1080\/1061186X.2019.1693576","article-title":"Serpina3n: Potential drug and challenges, mini review","volume":"28","author":"Aslam","year":"2020","journal-title":"J. Drug Target."},{"key":"2021113011191650600_B11","doi-asserted-by":"publisher","first-page":"1309","DOI":"10.1515\/hsz-2017-0180","article-title":"Kallistatin: double-edged role in angiogenesis, apoptosis and oxidative stress","volume":"398","author":"Chao","year":"2017","journal-title":"Biol. Chem."},{"key":"2021113011191650600_B12","doi-asserted-by":"publisher","first-page":"e210","DOI":"10.1161\/ATVBAHA.119.312413","article-title":"Protease Activity in Vascular Disease","volume":"39","author":"Slack","year":"2019","journal-title":"Arterioscler. Thromb. Vasc. Biol."},{"key":"2021113011191650600_B13","doi-asserted-by":"publisher","first-page":"1479","DOI":"10.1161\/ATVBAHA.119.313775","article-title":"Drug Targeting of Plasminogen Activator Inhibitor-1 Inhibits Metabolic Dysfunction and Atherosclerosis in a Murine Model of Metabolic Syndrome","volume":"40","author":"Khoukaz","year":"2020","journal-title":"Arterioscler. Thromb. Vasc. Biol."},{"key":"2021113011191650600_B14","doi-asserted-by":"publisher","first-page":"821","DOI":"10.1084\/jem.183.3.821","article-title":"Thrombin functions as an inflammatory mediator through activation of its receptor","volume":"183","author":"Cirino","year":"1996","journal-title":"J. Exp. Med."},{"key":"2021113011191650600_B15","doi-asserted-by":"publisher","first-page":"392","DOI":"10.1093\/cvr\/cvp066","article-title":"Is thrombin a key player in the \u2018coagulation-atherogenesis\u2019 maze?","volume":"82","author":"Borissoff","year":"2009","journal-title":"Cardiovasc. Res."},{"key":"2021113011191650600_B16","doi-asserted-by":"publisher","first-page":"258","DOI":"10.1038\/35025229","article-title":"Thrombin signalling and protease-activated receptors","volume":"407","author":"Coughlin","year":"2000","journal-title":"Nature"},{"key":"2021113011191650600_B17","first-page":"245","article-title":"Proteinase-activated receptors","volume":"53","author":"Macfarlane","year":"2001","journal-title":"Pharmacol. Rev."},{"key":"2021113011191650600_B18","doi-asserted-by":"publisher","first-page":"128","DOI":"10.1016\/j.abb.2016.06.018","article-title":"Role of heparin and non heparin binding serpins in coagulation and angiogenesis: A complex interplay","volume":"604","author":"Bhakuni","year":"2016","journal-title":"Arch. Biochem. Biophys."},{"key":"2021113011191650600_B19","doi-asserted-by":"publisher","first-page":"100961","DOI":"10.1016\/j.isci.2020.100961","article-title":"Serpina3c Regulates Adipogenesis by Modulating Insulin Growth Factor 1 and Integrin Signaling","volume":"23","author":"Choi","year":"2020","journal-title":"iScience"},{"key":"2021113011191650600_B20","doi-asserted-by":"publisher","first-page":"109745","DOI":"10.1016\/j.cellsig.2020.109745","article-title":"Serpina3c protects against high-fat diet-induced pancreatic dysfunction through the JNK-related pathway","volume":"75","author":"Ji","year":"2020","journal-title":"Cell. Signal."},{"key":"2021113011191650600_B21","doi-asserted-by":"publisher","first-page":"93","DOI":"10.1016\/j.neuroscience.2016.01.065","article-title":"A novel histochemical method for the visualization of thrombin activity in the nervous system","volume":"320","author":"Bushi","year":"2016","journal-title":"Neuroscience"},{"key":"2021113011191650600_B22","doi-asserted-by":"publisher","first-page":"2.9.1","DOI":"10.1002\/cpps.20","article-title":"Comparative Protein Structure Modeling Using MODELLER","volume":"86","author":"Webb","year":"2016","journal-title":"Curr. Protocols Protein Sci."},{"key":"2021113011191650600_B23","doi-asserted-by":"publisher","first-page":"291","DOI":"10.1146\/annurev.biophys.29.1.291","article-title":"Comparative protein structure modeling of genes and genomes","volume":"29","author":"Mart\u00ed-Renom","year":"2000","journal-title":"Annu. Rev. Biophys. Biomol. Struct."},{"key":"2021113011191650600_B24","doi-asserted-by":"publisher","first-page":"477","DOI":"10.1007\/BF00228148","article-title":"AQUA and PROCHECK-NMR: programs for checking the quality of protein structures solved by NMR","volume":"8","author":"Laskowski","year":"1996","journal-title":"J. Biomol. NMR"},{"key":"2021113011191650600_B25","doi-asserted-by":"publisher","first-page":"283","DOI":"10.1107\/S0021889892009944","article-title":"Procheck: a program to check the stereochemical quality of protein structures","volume":"26","author":"Laskowski","year":"1993","journal-title":"J. Appl. Crystallogr."},{"key":"2021113011191650600_B26","doi-asserted-by":"publisher","first-page":"11603","DOI":"10.1073\/pnas.0704409104","article-title":"Crystal structures of murine thrombin in complex with the extracellular fragments of murine protease-activated receptors PAR3 and PAR4","volume":"104","author":"Bah","year":"2007","journal-title":"Proc. Natl. Acad. Sci. U.S.A."},{"key":"2021113011191650600_B27","doi-asserted-by":"publisher","first-page":"3556","DOI":"10.1021\/acs.jcim.9b00445","article-title":"CoDockPP: A Multistage Approach for Global and Site-Specific Protein-Protein Docking","volume":"59","author":"Kong","year":"2019","journal-title":"J. Chem. Inf. Model."},{"key":"2021113011191650600_B28","doi-asserted-by":"publisher","first-page":"1100","DOI":"10.1002\/prot.25892","article-title":"Template-based modeling and ab-initio docking using CoDock in CAPRI","volume":"88","author":"Kong","year":"2020","journal-title":"Proteins"},{"key":"2021113011191650600_B29","doi-asserted-by":"publisher","first-page":"044130","DOI":"10.1063\/5.0014475","article-title":"Scalable molecular dynamics on CPU and GPU architectures with NAMD","volume":"153","author":"Phillips","year":"2020","journal-title":"J. Chem. Phys."},{"key":"2021113011191650600_B30","doi-asserted-by":"publisher","first-page":"863","DOI":"10.1038\/nsmb810","article-title":"The ternary complex of antithrombin-anhydrothrombin-heparin reveals the basis of inhibitor specificity","volume":"11","author":"Dementiev","year":"2004","journal-title":"Nat. Struct. Mol. Biol."},{"key":"2021113011191650600_B31","doi-asserted-by":"publisher","first-page":"e009562","DOI":"10.1161\/JAHA.118.009562","article-title":"Reduced Plasma Kallistatin Is Associated With the Severity of Coronary Artery Disease, and Kallistatin Treatment Attenuates Atherosclerotic Plaque Formation in Mice","volume":"7","author":"Yao","year":"2018","journal-title":"J. Am. Heart Assoc."},{"key":"2021113011191650600_B32","doi-asserted-by":"publisher","first-page":"473","DOI":"10.1016\/S0008-6363(98)00311-3","article-title":"Biphasic pattern of cell turnover characterizes the progression from fatty streaks to ruptured human atherosclerotic plaques","volume":"41","author":"Lutgens","year":"1999","journal-title":"Cardiovasc. Res."},{"key":"2021113011191650600_B33","doi-asserted-by":"publisher","first-page":"2266","DOI":"10.1172\/JCI117917","article-title":"Apoptosis of human vascular smooth muscle cells derived from normal vessels and coronary atherosclerotic plaques","volume":"95","author":"Bennett","year":"1995","journal-title":"J. Clin. Invest."},{"key":"2021113011191650600_B34","doi-asserted-by":"publisher","first-page":"575","DOI":"10.1002\/jcb.24693","article-title":"Recombinant human kallistatin inhibits angiogenesis by blocking VEGF signaling pathway","volume":"115","author":"Huang","year":"2014","journal-title":"J. Cell. Biochem."},{"key":"2021113011191650600_B35","doi-asserted-by":"publisher","first-page":"3245","DOI":"10.1182\/blood-2002-01-0185","article-title":"Kallistatin is a new inhibitor of angiogenesis and tumor growth","volume":"100","author":"Miao","year":"2002","journal-title":"Blood"},{"key":"2021113011191650600_B36","doi-asserted-by":"publisher","first-page":"e108859","DOI":"10.1371\/journal.pone.0108859","article-title":"SERPINA3K plays antioxidant roles in cultured pterygial epithelial cells through regulating ROS system","volume":"9","author":"Zhu","year":"2014","journal-title":"PLoS ONE"},{"key":"2021113011191650600_B37","doi-asserted-by":"publisher","first-page":"14","DOI":"10.1167\/iovs.14-14023","article-title":"Serine Proteinase Inhibitor SERPINA3K Suppresses Corneal Neovascularization via Inhibiting Wnt Signaling and VEGF","volume":"14","author":"Zhou","year":"2014","journal-title":"Invest Ophthalmol Vis Sci"},{"key":"2021113011191650600_B38","doi-asserted-by":"publisher","first-page":"24498","DOI":"10.1016\/S0021-9258(20)80553-5","article-title":"Kallistatin: a novel human serine proteinase inhibitor. Molecular cloning, tissue distribution, and expression in Escherichia coli","volume":"268","author":"Chai","year":"1993","journal-title":"J. Biol. Chem."},{"key":"2021113011191650600_B39","doi-asserted-by":"publisher","first-page":"43168","DOI":"10.1074\/jbc.M505598200","article-title":"The murine orthologue of human antichymotrypsin: a structural paradigm for clade A3 serpins","volume":"280","author":"Horvath","year":"2005","journal-title":"J. Biol. Chem."},{"key":"2021113011191650600_B40","doi-asserted-by":"publisher","first-page":"488","DOI":"10.1007\/s00239-004-2640-9","article-title":"Expression patterns of murine antichymotrypsin-like genes reflect evolutionary divergence at the Serpina3 locus","volume":"59","author":"Horvath","year":"2004","journal-title":"J. Mol. Evol."},{"key":"2021113011191650600_B41","doi-asserted-by":"crossref","first-page":"4378","DOI":"10.1182\/blood.V89.12.4378","article-title":"Thrombin generation by apoptotic vascular smooth muscle cells","volume":"89","author":"Flynn","year":"1997","journal-title":"Blood."},{"key":"2021113011191650600_B42","doi-asserted-by":"publisher","first-page":"513","DOI":"10.1093\/cvr\/cvs274","article-title":"Cyclic stretch-induced thrombin generation by rat vascular smooth muscle cells is mediated by the integrin \u03b1v\u03b23 pathway","volume":"96","author":"Mao","year":"2012","journal-title":"Cardiovasc. Res."},{"key":"2021113011191650600_B43","doi-asserted-by":"publisher","first-page":"3895","DOI":"10.2174\/138161210794454987","article-title":"Insulin, thrombine, ERK1\/2 kinase and vascular smooth muscle cells proliferation","volume":"16","author":"Isenovi\u0107","year":"2010","journal-title":"Curr. Pharm. Des."},{"key":"2021113011191650600_B44","doi-asserted-by":"publisher","first-page":"1594","DOI":"10.1161\/ATVBAHA.109.185801","article-title":"Thrombin induces EGF receptor expression and cell proliferation via a PKC(delta)\/c-Src-dependent pathway in vascular smooth muscle cells","volume":"29","author":"Hsieh","year":"2009","journal-title":"Arterioscler. Thromb. Vasc. Biol."},{"key":"2021113011191650600_B45","doi-asserted-by":"publisher","first-page":"2080","DOI":"10.1002\/jnr.24049","article-title":"The involvement of thrombin in the pathogenesis of glioblastoma","volume":"95","author":"Krenzlin","year":"2017","journal-title":"J. Neurosci. Res."}],"container-title":["Clinical Science"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/portlandpress.com\/clinsci\/article-pdf\/135\/3\/447\/903231\/cs-2020-1235.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/portlandpress.com\/clinsci\/article-pdf\/135\/3\/447\/903231\/cs-2020-1235.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2021,11,30]],"date-time":"2021-11-30T12:17:02Z","timestamp":1638274622000},"score":1,"resource":{"primary":{"URL":"https:\/\/portlandpress.com\/clinsci\/article\/135\/3\/447\/227612\/Protective-role-of-serpina3c-as-a-novel-thrombin"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,2,1]]},"references-count":45,"journal-issue":{"issue":"3","published-print":{"date-parts":[[2021,2,12]]}},"URL":"https:\/\/doi.org\/10.1042\/cs20201235","relation":{},"ISSN":["0143-5221","1470-8736"],"issn-type":[{"value":"0143-5221","type":"print"},{"value":"1470-8736","type":"electronic"}],"subject":[],"published-other":{"date-parts":[[2021,2]]},"published":{"date-parts":[[2021,2,1]]}}}