Soluble Thrombomodulin Purified from Human Urine Exhibits a Potent Anticoagulant Effect In Vitro and In Vivo

Summary We examined the anticoagulant activity of two major molecules of soluble thrombomodulin purified from human urine. The apparent molecular weights of these urinary thrombomodulins (UTMs) were 72,000 and 79,000, respectively. Both UTMs showed more potent cofactor activity for protein C activation [specific activity >5,000 thrombomodulin units (TMU)/mg] than human placental thrombomodulin (2,180 TMU/mg) and rabbit lung thrombomodulin (1,980 TMU/mg). The UTMs prolonged thrombin-induced fibrinogen clotting time (>1 TMU/ml), APTT (>5 TMU/ml), TT (>5 TMU/ml) and PT (>40 TMU/ml) in a dose-dependent fashion. These effects appeared in the concentration range of soluble thrombomodulins present in human plasma and urine. In the rat DIC model induced by thromboplastin, administration of UTMs by infusion (300-3,000 TMU/kg) restored the hematological abnormalities derived from DIC in a dose-dependent fashion. These results demonstrate that UTMs exhibit potent anticoagulant and antithrombotic activities, and could play a physiologically important role in microcirculation.

[1]  T. Tetaz,et al.  Purification and characterization of two forms of soluble thrombomodulin from human urine. , 1994, European journal of biochemistry.

[2]  I. Maruyama,et al.  Intravenous extended infusion of recombinant human soluble thrombomodulin prevented tissue factor‐induced disseminated intravascular coagulation in rats , 1994, American journal of hematology.

[3]  Y. Yazaki,et al.  Elevated serum and urinary thrombomodulin levels in patients with non-insulin-dependent diabetes mellitus. , 1994, Clinica chimica acta; international journal of clinical chemistry.

[4]  S. Yamamoto,et al.  Antithrombotic effect of recombinant human soluble thrombomodulin on endotoxin-induced disseminated intravascular coagulation in rats. , 1993, Thrombosis research.

[5]  S. Yamamoto,et al.  Urinary thrombomodulin, its isolation and characterization. , 1993, Journal of biochemistry.

[6]  K. Amano,et al.  Fluctuations in Plasma Levels of Thrombomodulin in Patients with DIC , 1992, Thrombosis and Haemostasis.

[7]  H. Niwano,et al.  Circulating thrombomodulin as a novel endothelial cell marker: Comparison of its behavior with von willebrand factor and tissue‐type plasminogen activator , 1992, American journal of hematology.

[8]  K. Nawa,et al.  The Glycosaminoglycan of Recombinant Human Soluble Thrombomodulin Affects Antithrombotic Activity in a Rat Model of Tissue Factor-Induced Disseminated Intravascular Coagulation , 1992, Thrombosis and Haemostasis.

[9]  J. Eidt,et al.  Intravenous recombinant soluble human thrombomodulin prevents venous thrombosis in a rat model. , 1991, Journal of vascular surgery.

[10]  F. Walker,et al.  Species specificity of the fibrinolytic effects of activated protein C. , 1991, Thrombosis research.

[11]  H. Araki,et al.  Inhibitory effects of activated protein C and heparin on thrombotic arterial occlusion in rat mesenteric arteries. , 1991, Thrombosis research.

[12]  S. Kimura,et al.  Plasma thrombomodulin in health and diseases. , 1990, Blood.

[13]  T. Ishikawa,et al.  Establishment of Enzyme Immunoassay of Human Thrombomodulin in Plasma and Urine Using Monoclonal Antibodies , 1990, Thrombosis and Haemostasis.

[14]  S. Yamamoto,et al.  Antithrombotic effect of recombinant human thrombomodulin on thrombin-induced thromboembolism in mice. , 1990, Blood.

[15]  William,et al.  Structure and function of thrombomodulin: a natural anticoagulant. , 1990, Blood.

[16]  P. Majerus,et al.  A role for thrombomodulin in the pathogenesis of thrombin-induced thromboembolism in mice. , 1988, Blood.

[17]  W. Lenz,et al.  An avidin-biotin ELISA for the detection of staphylococcal enterotoxins A and B. , 1986, Journal of immunological methods.

[18]  P. Majerus,et al.  Thrombomodulin is present in human plasma and urine. , 1985, The Journal of clinical investigation.

[19]  C. Bell,et al.  Thrombomodulin is found on endothelium of arteries, veins, capillaries, and lymphatics, and on syncytiotrophoblast of human placenta , 1985, The Journal of cell biology.

[20]  N. Aoki,et al.  Preparation of thrombomodulin from human placenta. , 1985, Thrombosis research.

[21]  C. Esmon,et al.  Isolation of a membrane-bound cofactor for thrombin-catalyzed activation of protein C. , 1982, The Journal of biological chemistry.

[22]  R. Marlar,et al.  Mechanism of action of human activated protein C, a thrombin-dependent anticoagulant enzyme. , 1982, Blood.

[23]  G. Kōsaki,et al.  FOY: [ethyl p-(6-guanidinohexanoyloxy) benzoate] methanesulfonate as a serine proteinase inhibitor. II. In vivo effect on coagulofibrinolytic system in comparison with heparin or aprotinin. , 1981, Thrombosis research.

[24]  C. Esmon,et al.  Identification of an endothelial cell cofactor for thrombin-catalyzed activation of protein C. , 1981, Proceedings of the National Academy of Sciences of the United States of America.

[25]  E. Davie,et al.  Preparation and properties of bovine factor VIII (antihemophilic factor). , 1980, Biochemistry.

[26]  U. K. Laemmli,et al.  Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4 , 1970, Nature.