Download PDF
Thromb Haemost 2006; 95(01): 128-133
DOI: 10.1160/TH05-06-0415
Cardiovascular Biology and Cell Signalling
Schattauer GmbH

Thrombomodulin plays an important role in arterial remodeling and neointima formation in mouse carotid ligation model

Yi-Heng Li
1   Department of Internal Medicine, College of Medicine, National Cheng Kung University, Tainan, Taiwan
,
Shu-Lin Liu
2   Department of Biochemistry and Molecular Biology, College of Medicine, National Cheng Kung University, Tainan, Taiwan
3   Institute of Basic Medical Sciences, College of Medicine, National Cheng Kung University, Tainan, Taiwan
,
Guey-Yueh Shi
2   Department of Biochemistry and Molecular Biology, College of Medicine, National Cheng Kung University, Tainan, Taiwan
3   Institute of Basic Medical Sciences, College of Medicine, National Cheng Kung University, Tainan, Taiwan
,
Guan-Hsiung Tseng
2   Department of Biochemistry and Molecular Biology, College of Medicine, National Cheng Kung University, Tainan, Taiwan
,
Ping-Yen Liu
1   Department of Internal Medicine, College of Medicine, National Cheng Kung University, Tainan, Taiwan
,
Hua-Lin Wu
2   Department of Biochemistry and Molecular Biology, College of Medicine, National Cheng Kung University, Tainan, Taiwan
3   Institute of Basic Medical Sciences, College of Medicine, National Cheng Kung University, Tainan, Taiwan
› Author AffiliationsFinancial support: This work was supported by grant 92–2314-B-006–131, 93–2314-B-006–094 from the National Science Council, Taipei, Taiwan, and 91-B-FA09–2–4 from the Ministry of Education Program for Promoting Academic Excellence of Universities, Taipei, Taiwan.
Further Information

Publication History

Received 14 June 2005

Accepted after resubmission 11 November 2005

Publication Date:
28 November 2017 (online)

    Permissions and Reprints

Summary

Thrombomodulin (TM) is a cell membrane-bound glycoprotein that functions as a thrombin cofactor in the activation of protein C. Its protein structure includes a N-terminal lectin-like domain (D1), 6 epidermal growth factor repeats (D2), a serine-threonine-rich region (D3), a transmembrane domain (D4) and a short cytoplasmic tail (D5). Recent studies have demonstrated the direct effect ofTM on cellular proliferation, adhesion and inflammation. In the study, we investigated the role of TM in vascular remodeling and neointima formation in a mouse carotid ligation model. TM expressions on the endothelium, neointima and media were examined in the ligated carotid artery by immunohistochemistry and quantitative real-time reverse transcription PCR. Endothelial TM expression decreased after ligation and appeared later in the media and neointima, which is quite similar to the appearance of TM in the human atherosclerotic process. Recombinant TMD123 was prepared. It was effective for thrombin-dependent protein C activation and the inhibition of leukocyte adhesion to the vessel wall after carotid ligation. RecombinantTMD123 and saline was administered immediately before and after carotid ligation. The TM-treated arteries demonstrated significantly less arterial dilatation (30279±12605 vs 73789±15073 µm2, p<0.05) in response to less neointima formation (14179±6538 vs 42227±8754 µm2, p<0.05) at 4 weeks after ligation. Our data indicated that there was a compensatory increase inTM expression in the media and neointima in relation to the reduced endothelial TM after carotid ligation. Early recombinant TM treatment in mice undergoing carotid ligation altered vascular remodeling and decreased the severity of neointima formation.

Keywords

Thrombomodulin - atherosclerosis - neointima
 

  • References

  • 1 Esmon CT. The regulation of natural anticoagulant pathways. Science 1987; 235: 1348-52.
    CrossrefPubMedGoogle Scholar
  • 2 Esmon CT. The roles of protein C and thrombomodulin in the regulation of blood coagulation. J Biol Chem 1989; 264: 4743-6.
    PubMedGoogle Scholar
  • 3 Wen D, Dittman WA, Ye RD. et al. Human thrombomodulin: complete cDNA sequence and chromosome localization of the gene. Biochemistry 1987; 26: 4350-7.
    CrossrefPubMedGoogle Scholar
  • 4 Kurosawa S, Stearns DJ, Jackson KW. et al. A 10-kDa cyanogen bromide fragment from the epidermal growth factor homology domain of rabbit thrombomodulin contains the primary thrombin binding site. J Biol Chem 1988; 263: 5993-6.
    PubMedGoogle Scholar
  • 5 Stearns DJ, Kurosawa S, Esmon CT. Microthrombomodulin. Residues 310–486 from the epidermal growth factor precursor homology domain of thrombomodulin will accelerate protein C activation. J Biol Chem 1988; 264: 3352-6.
    PubMedGoogle Scholar
  • 6 Li J, Garnette CS, Cahn M. et al. Recombinant thrombomodulin inhibits arterial smooth muscle cell proliferation induced by thrombin. J Vasc Surg 2000; 32: 804-13.
    CrossrefPubMedGoogle Scholar
  • 7 Tohda G, Oida K, Okada Y. et al Expression of thrombomodulin in atherosclerotic lesions and mitogenic activity of recombinant thrombomodulin in vascular smooth muscle cells. Arterioscler Thromb Vasc Biol 1998; 18: 1861-9.
    CrossrefPubMedGoogle Scholar
  • 8 Shi CS, Shi GY, Chang YS. et al. Evidence of human thrombomodulin domain as a novel angiogenic factor. Circulation 2005; 111: 1627-36.
    CrossrefPubMedGoogle Scholar
  • 9 Huang HC, Shi GY, Jiang SJ. et al. Thrombomodulin-mediated cell adhesion. J Biol Chem 2003; 278: 46750-9.
    CrossrefPubMedGoogle Scholar
  • 10 Conway EM, Van de Wouwer M, Pollefeyt S. et al. The lectin-like domain of thrombomodulin confers protection from neutrophil-mediated tissue damage by suppressing adhesion molecule expression via nuclear factor kB and mitogen-activated protein kinase pathways. J Exp Med 2002; 196: 565-77.
    CrossrefPubMedGoogle Scholar
  • 11 Li YH, Chen JH, Wu HL. et al. G-33A mutation in the promoter region of thrombomodulin gene and its association with coronary artery disease and plasma soluble thrombomodulin levels. Am J Cardiol 2000; 85: 8-12.
    PubMedGoogle Scholar
  • 12 Li YH, Chen CH, Yeh PS. et al. Functional mutation in the promoter region of thrombomodulin gene in relation to carotid atherosclerosis. Atherosclerosis 2001; 154: 713-9.
    CrossrefPubMedGoogle Scholar
  • 13 Yoshii Y, Okada Y, Sasaki S. et al. Expression of thrombomodulin in human aortic smooth muscle cells with special reference to atherosclerotic lesion types and age differences. Med Electron Microsc 2003; 36: 165-72.
    CrossrefPubMedGoogle Scholar
  • 14 Kumar A, Lindner V. Remodeling with neointima formation in the mouse carotid artery after cessation of blood flow. Arterioscler Thromb Vasc Biol 1997; 17: 2238-44.
    CrossrefPubMedGoogle Scholar
  • 15 Li J, Singh MJ, Itani M. et al. Recombinant human thrombomodulin inhibits arterial neointimal hyperplasia after balloon injury. J Vasc Surg 2004; 39: 1074-83.
    CrossrefPubMedGoogle Scholar
  • 16 Myers DL, Liaw L. Improved analysis of the vascular response to arterial ligation using a multivariate approach. Am J Pathol 2004; 164: 43-8.
    CrossrefPubMedGoogle Scholar
  • 17 Kumar A, Hoover JL, Simmons CA. et al. Remodeling and neointima formation in the carotid artery of normal and p-selectin-deficient mice. Circulation 1997; 96: 4333-42.
    CrossrefPubMedGoogle Scholar
  • 18 Faust SN, Levin M, Harrison OB. et al. Dysfunction of endothelial protein C activation in severe meningococcal sepsis. N Engl J Med 2001; 345: 408-16.
    CrossrefPubMedGoogle Scholar
  • 19 Conway EM, Rosenberg RD. Tumor necrosis factor suppresses transcription of the thrombomodulin gene in endothelial cells. Mol Cell Biol 1988; 08: 5588-92.
    CrossrefPubMedGoogle Scholar
  • 20 Nawroth PP, Handley DA, Esmon CT. et al. Interleukin 1 induces endothelial cell procoagulant while suppressing cell-surface anticoagulant activity. Proc Natl Acad Sci U S A 1986; 83: 3460-4.
    CrossrefPubMedGoogle Scholar
  • 21 Boehme MW, Deng Y, Raeth U. et al. Release of thrombomodulin from endothelial cells by concerted action of TNF-alpha and neutrophils: in vivo and in vitro studies. Immunology 1996; 87: 134-40.
    PubMedGoogle Scholar
  • 22 Laszik ZG, Zhou XJ, Ferrell GL. et al. Down-regulation of endothelial expression of endothelial cell protein C receptor and thrombomodulin in coronary atherosclerosis. Am J Pathol 2001; 159: 797-802.
    CrossrefPubMedGoogle Scholar
  • 23 Grinnell BW, Berg DT. Surface thrombomodulin modulates thrombin receptor responses on vascular smooth muscle cells. Am J Physio 1996; 270: H603-H609.
    PubMedGoogle Scholar
  • 24 Moll S, Lindley C, Pescatore S. et al. Phase I study of a novel recombinant human soluble thrombomodulin, ART-123. J Thromb Haemost 2004; 02: 1745-51.
    CrossrefPubMedGoogle Scholar
  • 25 Aoki Y, Takei R, Mohri M. et al. Antithrombotic effects of recombinant human soluble thrombomodulin (rhs-TM) on arteriovenous shunt thrombosis in rats. Am J Hematol 1994; 47: 162-6.
    CrossrefPubMedGoogle Scholar
  • 26 Mohri M, Gonda Y, Oka M. et al. The antithrombotic effects of recombinant human thrombomodulin (rhsTM) on tissue factor-induced disseminated intravascular coagulation in crab-eating monkeys (Macaca fascicularis). Blood Coagul Fibrinolysis 1997; 08: 274-83.
    CrossrefPubMedGoogle Scholar
  • 27 Riewald M, Petrovan RJ, Donner A. et al. Activation of endothelial cell protease activated receptor1 by the protein C pathway. Science 2002; 296: 1880-2.
    CrossrefPubMedGoogle Scholar
  • 28 Abeyama K, Stern DM, Ito Y. et al. The N-terminal domain of thrombomodulin sequesters high-mobolity-B1 protein, a novel antiinflammatory mechanism. J Clin Invest 2005; 115: 1267-74.
    CrossrefPubMedGoogle Scholar
  • 29 Sakaguchi T, Yan SF, Yan SD. et al. Central role of RAGE-dependent neointimal expansion in arterial restenosis. J Clin Invest 2003; 111: 959-72.
    CrossrefPubMedGoogle Scholar