title: Antithrombotic Effects of Nur77 and Nor1 Are Mediated Through Upregulating Thrombomodulin Expression in Endothelial Cells.
abstract: OBJECTIVE
  Thrombomodulin is highly expressed on the lumenal surface of vascular endothelial cells (ECs) and possesses potent anticoagulant, antifibrinolytic, and anti-inflammatory activities in the vessel wall. However, the regulation of thrombomodulin expression in ECs remains largely unknown.
  
  
  RESULTS
  In this study, we characterized nuclear receptor 4A family as a novel regulator of thrombomodulin expression in vascular ECs. We demonstrated that both nuclear receptors 4A, Nur77 and Nor1, robustly increase thrombomodulin mRNA and protein levels in human vascular ECs and in mouse liver tissues after adenovirus-mediated transduction of Nur77 and Nor1 cDNAs. Moreover, Nur77 deficiency and knockdown of Nur77 and Nor1 expression markedly attenuated the basal and vascular endothelial growth factor165-stimulated thrombomodulin expression. Mechanistically, we found that Nur77 and Nor1 increase thrombomodulin expression by acting through 2 different mechanisms. We showed that Nur77 barely affects thrombomodulin promoter activity, but significantly increases thrombomodulin mRNA stability, whereas Nor1 enhances thrombomodulin expression mainly through induction of Kruppel-like factors 2 and 4 in vascular ECs. Furthermore, we demonstrated that both Nur77 and Nor1 significantly increase protein C activity and inhibit tumor necrosis factor α-induced prothrombotic effects in human ECs. Deficiency of Nur77 increases susceptibility to arterial thrombosis, whereas enhanced expression of Nur77 and Nor1 protects mice from arterial thrombus formation.
  
  
  CONCLUSIONS
  Our results identified nuclear receptors 4A as novel regulators of thrombomodulin expression and function in vascular ECs and provided a proof-of-concept demonstration that targeted increasing expression of Nur77 and Nor1 in the vascular endothelium might represent a novel therapeutic approach for the treatment of thrombotic disorders.
authors.affiliation: From the Department of Pharmacy, Xinhua Hospital, Shanghai Jiaotong University, Shanghai, China (P.Y., X.W., J.Z.); Center for Translational Medicine, Department of Medicine, Thomas Jefferson University, Philadelphia, PA (P.Y., X.W., B.Y., G-.X.Z.,L.Y., J.S.); and Centers of Metabolic Disease Research and Cardiovascular Research, Department of Pharmacology, Temple University School of Medicine, Philadelphia, PA (X-.F.Y.).
authors.forename: Ping
authors.initials: P
authors.lastname: Yang
authors.affiliation: From the Department of Pharmacy, Xinhua Hospital, Shanghai Jiaotong University, Shanghai, China (P.Y., X.W., J.Z.); Center for Translational Medicine, Department of Medicine, Thomas Jefferson University, Philadelphia, PA (P.Y., X.W., B.Y., G-.X.Z.,L.Y., J.S.); and Centers of Metabolic Disease Research and Cardiovascular Research, Department of Pharmacology, Temple University School of Medicine, Philadelphia, PA (X-.F.Y.).
authors.forename: Xin
authors.initials: X
authors.lastname: Wei
authors.affiliation: From the Department of Pharmacy, Xinhua Hospital, Shanghai Jiaotong University, Shanghai, China (P.Y., X.W., J.Z.); Center for Translational Medicine, Department of Medicine, Thomas Jefferson University, Philadelphia, PA (P.Y., X.W., B.Y., G-.X.Z.,L.Y., J.S.); and Centers of Metabolic Disease Research and Cardiovascular Research, Department of Pharmacology, Temple University School of Medicine, Philadelphia, PA (X-.F.Y.).
authors.forename: Jian
authors.initials: J
authors.lastname: Zhang
authors.affiliation: From the Department of Pharmacy, Xinhua Hospital, Shanghai Jiaotong University, Shanghai, China (P.Y., X.W., J.Z.); Center for Translational Medicine, Department of Medicine, Thomas Jefferson University, Philadelphia, PA (P.Y., X.W., B.Y., G-.X.Z.,L.Y., J.S.); and Centers of Metabolic Disease Research and Cardiovascular Research, Department of Pharmacology, Temple University School of Medicine, Philadelphia, PA (X-.F.Y.).
authors.forename: Bing
authors.initials: B
authors.lastname: Yi
authors.affiliation: From the Department of Pharmacy, Xinhua Hospital, Shanghai Jiaotong University, Shanghai, China (P.Y., X.W., J.Z.); Center for Translational Medicine, Department of Medicine, Thomas Jefferson University, Philadelphia, PA (P.Y., X.W., B.Y., G-.X.Z.,L.Y., J.S.); and Centers of Metabolic Disease Research and Cardiovascular Research, Department of Pharmacology, Temple University School of Medicine, Philadelphia, PA (X-.F.Y.).
authors.forename: Guan-Xin
authors.initials: GX
authors.lastname: Zhang
authors.affiliation: From the Department of Pharmacy, Xinhua Hospital, Shanghai Jiaotong University, Shanghai, China (P.Y., X.W., J.Z.); Center for Translational Medicine, Department of Medicine, Thomas Jefferson University, Philadelphia, PA (P.Y., X.W., B.Y., G-.X.Z.,L.Y., J.S.); and Centers of Metabolic Disease Research and Cardiovascular Research, Department of Pharmacology, Temple University School of Medicine, Philadelphia, PA (X-.F.Y.).
authors.forename: Litian
authors.initials: L
authors.lastname: Yin
authors.affiliation: From the Department of Pharmacy, Xinhua Hospital, Shanghai Jiaotong University, Shanghai, China (P.Y., X.W., J.Z.); Center for Translational Medicine, Department of Medicine, Thomas Jefferson University, Philadelphia, PA (P.Y., X.W., B.Y., G-.X.Z.,L.Y., J.S.); and Centers of Metabolic Disease Research and Cardiovascular Research, Department of Pharmacology, Temple University School of Medicine, Philadelphia, PA (X-.F.Y.).
authors.forename: Xiao-Feng
authors.initials: XF
authors.lastname: Yang
authors.affiliation: From the Department of Pharmacy, Xinhua Hospital, Shanghai Jiaotong University, Shanghai, China (P.Y., X.W., J.Z.); Center for Translational Medicine, Department of Medicine, Thomas Jefferson University, Philadelphia, PA (P.Y., X.W., B.Y., G-.X.Z.,L.Y., J.S.); and Centers of Metabolic Disease Research and Cardiovascular Research, Department of Pharmacology, Temple University School of Medicine, Philadelphia, PA (X-.F.Y.). jianxin.sun@jefferson.edu.
authors.forename: Jianxin
authors.initials: J
authors.lastname: Sun
mesh_terms: D000818:Animals; D001777:Blood Coagulation; D016893:Carotid Stenosis / Q000235:genetics / Q000378:metabolism / Q000517:prevention & control*; D002478:Cells, Cultured; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D004195:Disease Models, Animal; D061307:Human Umbilical Vein Endothelial Cells / Q000378:metabolism*; D006801:Humans; D008297:Male; D008810:Mice, Inbred C57BL; D009419:Nerve Tissue Proteins / Q000235:genetics / Q000378:metabolism*; D057105:Nuclear Receptor Subfamily 4, Group A, Member 1 / Q000235:genetics / Q000378:metabolism*; D011401:Promoter Regions, Genetic; D011486:Protein C / Q000378:metabolism; D034622:RNA Interference; D020871:RNA Stability; D012333:RNA, Messenger / Q000378:metabolism; D011987:Receptors, Steroid / Q000235:genetics / Q000378:metabolism*; D011988:Receptors, Thyroid Hormone / Q000235:genetics / Q000378:metabolism*; D015398:Signal Transduction; D018180:Thrombomodulin / Q000235:genetics / Q000378:metabolism*; D013927:Thrombosis / Q000235:genetics / Q000378:metabolism / Q000517:prevention & control*; D013997:Time Factors; D014161:Transduction, Genetic; D014162:Transfection; D015854:Up-Regulation
chemical_list: D004268:DNA-Binding Proteins; C539821:NR4A1 protein, human; C490412:NR4A3 protein, human; D009419:Nerve Tissue Proteins; C539818:Nr4a1 protein, mouse; C490411:Nr4a3 protein, mouse; D057105:Nuclear Receptor Subfamily 4, Group A, Member 1; D011486:Protein C; D012333:RNA, Messenger; D011987:Receptors, Steroid; D011988:Receptors, Thyroid Hormone; C546931:THBD protein, human; C569169:THBD protein, mouse; D018180:Thrombomodulin
keywords: anticoagulants; endothelial cells; thrombomodulin; thrombosis; tumor necrosis factor