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Thromb Haemost 1981; 45(03): 225-229
DOI: 10.1055/s-0038-1650175
Original Article
Schattauer GmbH Stuttgart

Comparison of the Relative Fibrinogenolytic, Fibrinolytic and Thrombolytic Properties of Tissue Plasminogen Activator and Urokinase in Vitro

O Matsuo
The Center for Thrombosis and Vascular Research, Department of Medical Research, University of Leuven, Belgium
,
D C Rijken
The Center for Thrombosis and Vascular Research, Department of Medical Research, University of Leuven, Belgium
,
D Collen
The Center for Thrombosis and Vascular Research, Department of Medical Research, University of Leuven, Belgium
› Author Affiliations
Further Information

Publication History

Received 24 December 1980

Accepted 08 March 1981

Publication Date:
06 July 2018 (online)

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Summary

The relative fibrinogenolytic, fibrinolytic and thrombolytic properties of human tissue plasminogen activator and human urokinase were compared in purified systems, in whole human plasma and in a system composed of a radioactive human blood clot (<sup>125</sup>I-fibrinogen) hanging in circulating human plasma. The human tissue plasminogen activator was highly purified from the culture fluid of a human melanoma cell line.

In purified systems composed of fibrinogen or fibrin, plasminogen and α<sub>2</sub>-antiplasmin as well as in whole plasma, tissue plasminogen activator digested fibrin without degrading fibrinogen significantly. Urokinase did not have this specific fibrinolytic effect.

In the circulating plasma system, the degree of fibrinolysis was proportional to the amount of activator added, tissue plasminogen activator being about 10 times more efficient than urokinase. In addition, tissue plasminogen activator appeared to cause negligible fibrinogen degradation. Tissue plasminogen activator still induced significant thrombolysis at a concentration of 10 IU per ml whereas no effect of urokinase was observed at 20 IU per ml. Infusion of 100 IU (1 (μg) of tissue plasminogen activator per ml resulted in moderate activation of the fibrinolytic system as judged from a decrease of plasminogen and α<sub>2</sub>-antiplasmin to 40-50 percent. Nevertheless, extensive fibrinolysis (50 to 80 percent of radioactivity released after 12 hrs) and only very limited fibrinogenolysis were observed. An equivalent amount of urokinase (100 IU per ml) only induced approximately 15 percent lysis in 12 hrs. At higher concentrations of urokinase (260 IU per ml or more) extensive activation of the fibrinolytic system was obtained as evidenced by a depletion of plasminogen, α<sub>2</sub>-antiplasmin and fibrinogen. This was associated with extensive fibrinolysis (approximately 60 percent after 12 hrs). It is concluded that human tissue plasminogen activator is a more specific and effective fibrinolytic-thrombolytic agent than human urokinase.

Keywords

Tissue plasminogen activator - Urokinase - Fibrinogenolysis - Fibrinolysis - Thrombolysis
 

  • References

  • 1 Wiman B, Collen D. Molecular mechanism of physiological fibrinolysis. Nature 1978; 272: 549-550
    CrossrefPubMedGoogle Scholar
  • 2 Collen D, Verstraete M. α2-Antiplasmin consumption and fibrinogen breakdown during thrombolytic therapy. Thromb Res 1979; 14: 631-739
    CrossrefPubMedGoogle Scholar
  • 3 Rijken DC, Wijngaards G, Zaal-De JongM, Welbergen J. Purification and partial characterization of plasminogen activator from human uterine tissue. Biochim Biophys Acta 1979; 580: 140-153
    CrossrefPubMedGoogle Scholar
  • 4 Rijken DC, Collen D. Purification and characterization of the plasminogen activator secreted by human melanoma cells in culture. J Biol Chem. 1981. (in press)
    Google Scholar
  • 5 Rijken DC, Wijngaards G, Welbergen J. Relationship between tissue plasminogen activator and the activator in blood and vascular wall. Thromb Res 1980; 18: 815-830
    CrossrefPubMedGoogle Scholar
  • 6 Rijken DC, Wijngaards G, Welbergen J. Immunological characterization of plasminogen activator activities in human tissues and body fluids. J Lab Clin Med. 1981. (in press)
    Google Scholar
  • 7 Blombäck B, Blombäck M. Purification of human and bovine fibrinogen. Arkiv för Kemi 1956; 10: 415-428
    PubMedGoogle Scholar
  • 8 McFarlane AS. Efficient trace-labelling of proteins with iodine. Nature 1958; 182: 53
    PubMedGoogle Scholar
  • 9 Deutsch DG, Mertz ET. Plasminogen: purification from human plasmin by affinity chromatography. Science 1970; 170: 1095-1096
    CrossrefPubMedGoogle Scholar
  • 10 Collen D, Ong EB, Johnson AJ. Human plasminogen: in vitro and in vivo evidence for the biological integrity of NH2-terminal glutamic acid plasminogen. Thromb Res 1975; 07: 515-529
    CrossrefPubMedGoogle Scholar
  • 11 Wiman B, Collen D. Purification and characterization of human antiplasmin, the fast-acting plasmin inhibitor in plasma. Eur J Biochem 1977; 78: 19-26
    CrossrefPubMedGoogle Scholar
  • 12 Fenton JW, Fascon MJ, Stackrow AB, Aronson DL, Young AM, Finlayson JS. Human thrombins. Production, evaluation and properties of α-thrombin. J Biol Chem 1977; 252: 3587-3598
    PubMedGoogle Scholar
  • 13 Vermylen C, De VrekerR, Verstraete M. A rapid enzymatic method for assay of fibrinogen fibrin polymerization time (FPT-test). Clin Chim Acta 1963; 08: 418-424
    CrossrefPubMedGoogle Scholar
  • 14 Merskey C, Lalezari I, Johnson AJ. A rapid, simple, sensitive method for measuring fibrinolytic split products in human serum. Proc Soc Exp Biol Med 1969; 131: 871-875
    CrossrefPubMedGoogle Scholar
  • 15 Friberger P, Knös M. Plasminogen determination in human plasma. In: “Chromogenic peptide substrates”. Scully MF, Kakkar VV. eds Churchill Livingstone; Edinburgh: 1979. p 128-140
    Google Scholar
  • 16 Edy J, De CockF, Collen D. Inhibition of plasmin by normal and antiplasmin-depleted human plasma. Thromb Res 1976; 08: 513-518
    CrossrefPubMedGoogle Scholar
  • 17 Vermylen C, Verstraete M. Antithrombin V: critical evaluation of its assessment and properties. Thromb Diath haemorrh 1960; 05: 267-284
    PubMedGoogle Scholar
  • 18 Todd AS. Localization of fibrinolytic activity in tissues. Brit Med Bull 1964; 20: 210-212
    CrossrefPubMedGoogle Scholar
  • 19 Vetterlein D, Young PL, Bell TE, Roblin R. Immunological characterization of multiple molecular weight forms of human cell plasminogen activators. J Biol Chem 1979; 254: 575-578
    PubMedGoogle Scholar
  • 20 Bernik MB, Rijken DC, Wijngaards G. Production of two immunologically distinct plasminogen activators by human tissue in culture. Thromb Haemost 1979; 42: 414
    PubMedGoogle Scholar
  • 21 Wilson EL, Becker MLB, Hoal EG, Dowdle EB. Molecular species of plasminogen activators secreted by normal and neoplastic human cells. Cancer Res 1980; 40: 933-938
    PubMedGoogle Scholar
  • 22 Gurewich V, Hyde E, Upinski B. The resistance of fibrinogen and soluble fibrin monomer in blood to degradation by a potent plasminogen activator derived from cadaver limbs. Blood 1975; 46: 555-565
    PubMedGoogle Scholar
  • 23 Camiolo SM, Thorsen S, Astrup T. Fibrinogenolysis and fibrinolysis with tissue plasminogen activator, urokinase, streptokinase-activated human globulin, and plasmin. Proc Soc Exp Med 1971; 138: 277-280
    CrossrefPubMedGoogle Scholar
  • 24 Thorsen S, Glas-Greenwalt P, Astrup T. Differences in the binding to fibrin of urokinase and tissue plasminogen activator. Thromb Diath haemorrh 1972; 28: 65-74
    PubMedGoogle Scholar
  • 25 Wallen P. Chemistry on plasminogen and plasminogen activation. In: “Progress in chemical fibrinolysis and thrombolysis, vol 3”. Davidson JF, Rowan RM, Samama MM, Desnoyers PC. eds Raven Press; New York: 1978. p 167-181
    Google Scholar
  • 26 Wallen P. Activation of plasminogen with urokinase and tissue activator. In: “Thrombosis and urokinase”. Paoletti R, Sherry S. eds Academic Press; New York: 1977. p 091-102
    Google Scholar
  • 27 Thorsen S. >Differences in the binding to fibrin of native plasminogen and plasminogen modified by proteolytic degradation. Influence of ω-aminocarboxylic adds. Biochim Biophys Acta 1975; 393: 55-65
    CrossrefPubMedGoogle Scholar
  • 28 Wiman B, Wallén P. On the specific function of the lysine binding site in human plasminogen. Thromb Res 1977; 10: 213-222
    CrossrefPubMedGoogle Scholar
  • 29 Rákóczi I, Wiman B, Collen D. On the biological significance of the specific interaction between fibrin, plasminogen and antiplasmin. Biochim Biophys Acta 1978; 540: 295-300
    CrossrefPubMedGoogle Scholar
  • 30 Christensen U, Clemmensen I. Kinetic properties of theprimaryinhibitor of plasmin from human plasma. Biochem J 1977; 163: 389-391
    CrossrefPubMedGoogle Scholar
  • 31 Wiman B, Collen D. On the kinetics of the reaction between human antiplasmin and plasmin. Eur J Biochem 1978; 84: 573-578
    CrossrefPubMedGoogle Scholar
  • 32 Matsuo O, Rijken DC, Collen D. Thrombolytic properties of human tissue plasminogen activator, as compared to urokinase in rabbits with an experimental pulmonary embolus. Nature. 1981. (in press)
    Google Scholar