Thromb Haemost 1994; 71(04): 507-510
DOI: 10.1055/s-0038-1642468
Review Article
Schattauer GmbH Stuttgart

Defibrotide Stimulates Expression of Thrombomodulin in Human Endothelial Cells

Quansheng Zhou
The Jiangsu institute of Hematology, Thrombosis and Haemostasis Research Unit, Suzhou Medical College, Suzhou, China
,
Xiaohong Chu
The Jiangsu institute of Hematology, Thrombosis and Haemostasis Research Unit, Suzhou Medical College, Suzhou, China
,
Changgeng Ruan
The Jiangsu institute of Hematology, Thrombosis and Haemostasis Research Unit, Suzhou Medical College, Suzhou, China
› Author Affiliations
Further Information

Publication History

Received: 09 August 1993

Accepted after resubmission: 21 December 1993

Publication Date:
06 July 2018 (online)

Summary

Cultured human umbilical vein endothelial cells were incubated with defibrotide at concentrations of 0, 5, 50 and 500 jxg/ml for 4 and 24 h respectively. Thrombomodulin activity and molecules on the surface of the cells were determined by chromogenic assay and radioimmunoassay, thrombomodulin antigen in endothelial cells and in conditioned medium of the cells was measured by immunoradioassay. Thrombomodulin mRNA within the cells was analysed by slot blot. After 24 h of incubation, the activity and molecules of thrombomodulin on the surface of endothelial cells, as well as the antigen and mRNA of thrombomodulin in the cells were significantly increased in a dose dependent manner. However, the level of thrombomodulin antigen in conditioned medium was about equal to that of the control. Our data indicate that defibrotide stimulates expression of thrombomodulin in human endothelial cells. These beneficial effects may play a role in antithrombotic activity of defibrotide.

 
  • References

  • 1 Niada R, Mantovani M, Prino G, Pescador R, Nardi G. Antithrombotic activity of polydeoxyribonucleotides of mammalian organ (laboratory code: Fraction P) in experimental animals. Thromb Haemostas 1979; 42: 474 (Abstr)
  • 2 Pescador R, Mantovani M, Prino G, Madonna M. Pharmacokinetics of defibrotide and of its profibrinolytic activity in the rabbit. Thromb Res 1983; 30: 1-11
  • 3 Mussoni L, Evolvi C, Donati MB. Activation of plasma and vascular fibrinolytic activity by a polydeoxyribonucleotidicsubstance Fraction P. in rats and rabbits. Thromb Haemostas 1979; 42: 388 (Abstr)
  • 4 Grodzinska L, Konig E, Schror K. Defibrotide is equipotent to urokinase in stimulating arterial and venous thrombolysis. Pharmacol Res Commun 1987; 19: 609-615
  • 5 Niada R, Pescador R, Porta R, Mantovani M, Prino G. Defibrotide is antithrombotic and thrombolytic against rabbit venous thrombosis. Haemostasis 1986; 16 (Suppl 1) 3-8
  • 6 Niada R, Porta R, Pescador R, Mantovani M, Prino G. Cardioprotective effects of defibrotide in acute myocardial ischemia in the cat. Thromb Res 1985; 38: 71-81
  • 7 Niada R, Porta R, Pescador R, Mantovani M, Prino G. Protective activity of defibrotide against lethal acute myocardial ischemia in the cat. Thromb Res 1986; 42: 363-374
  • 8 Thiermermann C, Lobel P, Schror K. Usefulness of defibrotide in protecting ischemic myocardium from early reperfusion damage. Am J Cardiol 1985; 56: 978-982
  • 9 Thiermermann C, Thomas GR, Vane JR. Defibrotide reduces infarct size in a rabbit model of experimental myocardial ischaemia and reperfusion. Br J Pharmacol 1989; 97: 401-408
  • 10 Berti F, Rossoni G, Niada R, Folco G, Omini C, Tondo C. Defibrotide, an antithrombotic substance that preserves postsynaptic alfa and beta-adrenergic function in post acute infarcted rabbit hearts. J Cardiovasc Pharmacol 1986; 8: 235-240
  • 11 Berti F, Rossoni G, Omini C, Folco G, Daffonchio L, Vigano T, Tondo C. Defibrotide, an antithrombotic substance which prevents myocardial contracture in ischemic rabbit heart. Eur J Pharmacol 1987; 135: 375-382
  • 12 Pescador R, Porta R, Mantovani M, Prino G. Cardioprotective effects of defibrotide: A minireview. Semin Thromb Hemost 1991; 17 (Suppl 1) 96-100
  • 13 Niada R, Mantovani M, Prino G, Pescador R, Berti F, Omini C, Folco GC. Antithrombotic activity of a polydeoxyribonucleotidic substance extracted from mammalian organs: a possible link with prostacyclin. Thromb Res 1981; 23: 233-46
  • 14 Niada R, Mantovani M, Prino G, Pescador R, Porta R, Berti F, Folco GC, Omini C, Vigano T. PGI2-generation and antithrombotic activity of orally administered defibrotide. Pharmacol Res Commun 1982; 14: 949-957
  • 15 Lobel P, Schror K. Selective stimulation of coronary vascular PGI2 but not of platelet thromboxane formation by defibrotide in the platelet perfused heart. Naunyn-Schmiedebergs Arch Pharmacol 1988; 331: 125-130
  • 16 Cornelli U, Nazzari M. Defibrotide: An orverview of clinical pharmacology and early clinical studies. Semin Thromb Hemost 1988; 14 (Suppl 1): 64-70
  • 17 Violi F, Ferro D, Alessandri C, Quintarelli C, Saliola M, Balsano F. Inhibition of tissue plasminogen activator inhibitor by defibrotide in atherosclerotic patients. Semin Thromb Hemost 1989; 15: 226-229
  • 18 Patrassi GM, Sartori MT, Viero ML, Scapinello MP, Boeri G, Gi- rolami A. Fibrinolytic effects of defibrotide in atherosclerotic patients. Semin Thromb Hemost 1991; 17 (Suppl 1): 101-105
  • 19 Diperri T, Laghi Pasini F, Frigerio C, Capecchi P, Messa G, Franchi M. Usefulness of defibrotide in the treatment of vascular disorders: A clinical- pharmacologic study. Semin Thromb Hemost 1991; 17 (Suppl 1): 85-95
  • 20 Milazzotto F, Carelli M, Citone C, Marcitullio G, Giampaolo P, Malinconico U. Use of defibrotide in the treatment of acute myocardial infarction. Semin Thromb Hemost 1989; 15: 464-469
  • 21 Mattioli G, Cappello C, Fusaro MT. Treatment of acute myocardial infarction with defibrotide. Semin Thromb Hemost 1989; 15: 470-473
  • 22 Niada R, Porta R, Pescador R, Mantovani M, Prino G. Thrombolytic activity of defibrotide apainsi old venous thrombi. Semin Thromb Hemost 1989; 15: 474-479
  • 23 Esmon CT, Owen WG. Identification of an endothelial cells cofactor for thrombin-catalyzed activation of protein C. Proc Natl Acad Sci USA 1981; 78: 2249-2252
  • 24 Dittman WA, Majerus PW. Structure and function of thrombomodulin: A natural anticoagulant. Bluud 1990; 75: 329-336
  • 25 Brosstad F. Purification, characterization and insolubilization of bovine thrombin. Thromb Res 1977; 11: 119-130
  • 26 Stenflo J. A new vitamin K-dependent protein. J Biol Chem 1976; 251: 355-363
  • 27 Jaffe EA, Nachman RL, Becker CG, Minick CR. Culture of human endothelial cells derived from umbilical cords: Identification by morphological and immunologic criteria. J Clin Invest 1973; 52: 2745-2756
  • 28 Chomczynski P, Szcchi N. Single-step method of RNA isolation by acid guanidium thiocynate-phenolchloroform extraction. Anal Biochem 1987; 162: 156-159
  • 29 Zhou QS, Zhao YM, Li P, Bai X, Ruan CG. A monoclonal antibody (SZ- 53) against thrombomodulin inhibits thrombin-mediated release of t-PA and PGI2 from endothelial cells. Nouv Rew Fr Hematol 1991; 33: 281-286
  • 30 Zhou QS, Zhao YM, Bai X, Ruan CG. Establishment of a technique of radioimmunoassay for measurement of thrombomodulin on the surface of endothelial cells. Chin J Hematol 1991; 19: 489-490
  • 31 Zhou QS, Zaho YM, Xu CS, Yu Z, Yao DY, Gao YM, Ruan CG. Increase in plasma thrombomodulin and decrease in plasma von Willebrand factor after regular radiotherapy of patients with cancer. Thromb Res 1992; 68: 109-118
  • 32 Feinberg AP, Vogelstein B. A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem 1983; 132: 6-13
  • 33 Lentz SR, Sadler JE. Inhibition of thrombomodulin surface expression and protein C activation by thrombogenic agent homocysteine. J Clin Invest 1991; 88: 1906-14
  • 34 Garcia MM, Kaufman CE, Comp PC. Decrease in endothelial cell-dependent protein C activation induced by treatment with cyclosporine. Transplantation 1991; 51: 701-705
  • 35 Archipoff G, Beretz A, Freyssinet JM, Klein-Soyer C, Brisson C, Cazenave JP. Heterogeneous regulation of constitutive thrombomodulin or inducible tissue-factor activities on the surface of human saphenous vein endothelial cells in culture following stimulation by interleukin-1, tumor necrosis factor, thrombin orphorbol ester. Biochem J 1991; 273: 679-684
  • 36 Ishii H, Kiznlci K, Uchiyama H, Horic S, Kazama M. Cyclic AMP in creases thrombomodulin expression on membrane surface of cultured human umbilical vein endothelial cell. Thromb Res 1990; 59: 841-850
  • 37 Hirokawa K, Aoki N. Regulatory mechanisms for thrombomodulin expression in human umbilical vein endothelial cells in vivo. J Cell Physiol 1991; 147: 157-165
  • 38 Horie S, Kizaki K, Ishn H, Kazama M. Retinoic acid stimulates expression of thrombomodulin a cell surface anticoagulant glycoprotein on human endothelial cells Differences between up-regulation of thrombomodulin by retinoic acid and cyclic AMP. Biochem J 1992; 281: 149-154
  • 39 Uchiyama H, Hiraishi S, Ohtani H, Ishii H, Kazama M. Plasma thrombomodulin is originated by damage of endothelial cell. Thromb Haemostas 1989; 62: 276
  • 40 Zhou QS, Zhao YM, Li PX, Bai X, Ruan C. Thrombomodulin as a marker of radiation-induced endothelial cell injury. Radiat Res 1992; 131: 285-289
  • 41 Zhou QS, Xiaohong Chu, Xiaodong Wang, Fugang Li, Jianming Gu. Changgeng Ruan. Studies on the preparation and application of thrombomodulin gene probe. Chinese J Exp Hematol. 1993. (Submitted)