Skip to main content
NCBI home page
The Journal of Clinical Investigation logoLink to The Journal of Clinical Investigation
. 1989 Aug;84(2):586–591. doi: 10.1172/JCI114202

Recombinant plasminogen activator inhibitor-1 reverses the bleeding tendency associated with the combined administration of tissue-type plasminogen activator and aspirin in rabbits.

D E Vaughan 1, P J Declerck 1, M De Mol 1, D Collen 1
PMCID: PMC548919  PMID: 2503540

Abstract

The major side effect of thrombolytic therapy is bleeding; however, the pathogenesis of this potential complication is not well understood. Accordingly, we examined the effects of aspirin and recombinant human tissue-type plasminogen activator (rt-PA) on serial template bleeding times and on hemostasis parameters in rabbits. The administration of intravenous aspirin (15 mg/kg) produced a slight prolongation in bleeding times, from 2.1 +/- 0.5 to 2.6 +/- 0.5 min (mean +/- SD, n = 26, P less than 0.01), whereas rt-PA (1 mg/kg per h for 2 h) lengthened the bleeding time from 2.4 +/- 0.3 to 3.2 +/- 0.6 min (n = 5, P = NS). Combination of aspirin with 0.5 mg/kg per h of rt-PA for 2 h prolonged the bleeding time from 2.5 +/- 0.4 to 6.2 +/- 0.9 min (n = 10, P less than 0.01), with an associated fibrinogen decrease of approximately 15%. The combination of aspirin with 1 mg/kg per h of rt-PA for 2 h prolonged the bleeding time from 3.0 +/- 0.3 to 8.3 +/- 1.4 min (n = 8, P less than 0.01) and simultaneously induced a decrease of plasma fibrinogen by approximately 40%. Virtually all animals treated with rt-PA and aspirin manifested a bleeding tendency, as evidenced by spontaneous rebleeding at sites of previously performed template bleeding times or oozing at the femoral venous catheterization site. Intravenous bolus injection of 1 mg/kg of guanidine hydrochloride-reactivated recombinant human plasminogen activator inhibitor-1 (rPAI-1) at the end of the rt-PA infusion resulted in complete reversal, within 5 min, of the prolongation of the bleeding time, and in a disappearance of the bleeding tendency. Nonreactivated rPAI-1 and tranexamic acid were significantly less potent in reversing the bleeding time prolongation. These findings indicate that aspirin and rt-PA given separately do not markedly affect the template bleeding time, but in combination induce a marked prolongation associated with a significant bleeding tendency. This bleeding time prolongation can be rapidly normalized by the administration of reactivated rPAI-1.

Full text

PDF
586

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Alessi M. C., Declerck P. J., De Mol M., Nelles L., Collen D. Purification and characterization of natural and recombinant human plasminogen activator inhibitor-1 (PAI-1). Eur J Biochem. 1988 Aug 15;175(3):531–540. doi: 10.1111/j.1432-1033.1988.tb14225.x. [DOI] [PubMed] [Google Scholar]
  2. Andreasen P. A., Riccio A., Welinder K. G., Douglas R., Sartorio R., Nielsen L. S., Oppenheimer C., Blasi F., Danø K. Plasminogen activator inhibitor type-1: reactive center and amino-terminal heterogeneity determined by protein and cDNA sequencing. FEBS Lett. 1986 Dec 15;209(2):213–218. doi: 10.1016/0014-5793(86)81113-9. [DOI] [PubMed] [Google Scholar]
  3. CLAUSS A. Gerinnungsphysiologische Schnellmethode zur Bestimmung des Fibrinogens. Acta Haematol. 1957 Apr;17(4):237–246. doi: 10.1159/000205234. [DOI] [PubMed] [Google Scholar]
  4. Califf R. M., Topol E. J., George B. S., Boswick J. M., Abbottsmith C., Sigmon K. N., Candela R., Masek R., Kereiakes D., O'Neill W. W. Hemorrhagic complications associated with the use of intravenous tissue plasminogen activator in treatment of acute myocardial infarction. Am J Med. 1988 Sep;85(3):353–359. doi: 10.1016/0002-9343(88)90586-4. [DOI] [PubMed] [Google Scholar]
  5. Collen D., Bounameaux H., De Cock F., Lijnen H. R., Verstraete M. Analysis of coagulation and fibrinolysis during intravenous infusion of recombinant human tissue-type plasminogen activator in patients with acute myocardial infarction. Circulation. 1986 Mar;73(3):511–517. doi: 10.1161/01.cir.73.3.511. [DOI] [PubMed] [Google Scholar]
  6. Collen D., Stassen J. M., Verstraete M. Thrombolysis with human extrinsic (tissue-type) plasminogen activator in rabbits with experimental jugular vein thrombosis. Effect of molecular form and dose of activator, age of the thrombus, and route of administration. J Clin Invest. 1983 Feb;71(2):368–376. doi: 10.1172/JCI110778. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Declerck P. J., Alessi M. C., Verstreken M., Kruithof E. K., Juhan-Vague I., Collen D. Measurement of plasminogen activator inhibitor 1 in biologic fluids with a murine monoclonal antibody-based enzyme-linked immunosorbent assay. Blood. 1988 Jan;71(1):220–225. [PubMed] [Google Scholar]
  8. Declerck P. J., De Mol M., Alessi M. C., Baudner S., Pâques E. P., Preissner K. T., Müller-Berghaus G., Collen D. Purification and characterization of a plasminogen activator inhibitor 1 binding protein from human plasma. Identification as a multimeric form of S protein (vitronectin). J Biol Chem. 1988 Oct 25;263(30):15454–15461. [PubMed] [Google Scholar]
  9. Fitzgerald D. J., Catella F., Roy L., FitzGerald G. A. Marked platelet activation in vivo after intravenous streptokinase in patients with acute myocardial infarction. Circulation. 1988 Jan;77(1):142–150. doi: 10.1161/01.cir.77.1.142. [DOI] [PubMed] [Google Scholar]
  10. Ginsburg D., Zeheb R., Yang A. Y., Rafferty U. M., Andreasen P. A., Nielsen L., Dano K., Lebo R. V., Gelehrter T. D. cDNA cloning of human plasminogen activator-inhibitor from endothelial cells. J Clin Invest. 1986 Dec;78(6):1673–1680. doi: 10.1172/JCI112761. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Guccione M. A., Kinlough-Rathbone R. L., Packham M. A., Harfenist E. J., Rand M. L., Greenberg J. P., Perry D. W., Mustard J. F. Effects of plasmin on rabbit platelets. Thromb Haemost. 1985 Feb 18;53(1):8–14. [PubMed] [Google Scholar]
  12. Hajjar K. A., Hamel N. M., Harpel P. C., Nachman R. L. Binding of tissue plasminogen activator to cultured human endothelial cells. J Clin Invest. 1987 Dec;80(6):1712–1719. doi: 10.1172/JCI113262. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Hamsten A., Wiman B., de Faire U., Blombäck M. Increased plasma levels of a rapid inhibitor of tissue plasminogen activator in young survivors of myocardial infarction. N Engl J Med. 1985 Dec 19;313(25):1557–1563. doi: 10.1056/NEJM198512193132501. [DOI] [PubMed] [Google Scholar]
  14. Hekman C. M., Loskutoff D. J. Endothelial cells produce a latent inhibitor of plasminogen activators that can be activated by denaturants. J Biol Chem. 1985 Sep 25;260(21):11581–11587. [PubMed] [Google Scholar]
  15. Holvoet P., Cleemput H., Collen D. Assay of human tissue-type plasminogen activator (t-PA) with an enzyme-linked immunosorbent assay (ELISA) based on three murine monoclonal antibodies to t-PA. Thromb Haemost. 1985 Oct 30;54(3):684–687. [PubMed] [Google Scholar]
  16. Kruithof E. K., Tran-Thang C., Ransijn A., Bachmann F. Demonstration of a fast-acting inhibitor of plasminogen activators in human plasma. Blood. 1984 Oct;64(4):907–913. [PubMed] [Google Scholar]
  17. Lambers J. W., Cammenga M., König B. W., Mertens K., Pannekoek H., van Mourik J. A. Activation of human endothelial cell-type plasminogen activator inhibitor (PAI-1) by negatively charged phospholipids. J Biol Chem. 1987 Dec 25;262(36):17492–17496. [PubMed] [Google Scholar]
  18. Lawrence D. A., Loskutoff D. J. Inactivation of plasminogen activator inhibitor by oxidants. Biochemistry. 1986 Oct 21;25(21):6351–6355. doi: 10.1021/bi00369a001. [DOI] [PubMed] [Google Scholar]
  19. Loscalzo J., Braunwald E. Tissue plasminogen activator. N Engl J Med. 1988 Oct 6;319(14):925–931. doi: 10.1056/NEJM198810063191407. [DOI] [PubMed] [Google Scholar]
  20. Loscalzo J., Vaughan D. E. Tissue plasminogen activator promotes platelet disaggregation in plasma. J Clin Invest. 1987 Jun;79(6):1749–1755. doi: 10.1172/JCI113015. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Niewiarowski S., Senyi A. F., Gillies P. Plasmin-induced platelet aggregation and platelet release reaction. Effects on hemostasis. J Clin Invest. 1973 Jul;52(7):1647–1659. doi: 10.1172/JCI107345. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Ny T., Sawdey M., Lawrence D., Millan J. L., Loskutoff D. J. Cloning and sequence of a cDNA coding for the human beta-migrating endothelial-cell-type plasminogen activator inhibitor. Proc Natl Acad Sci U S A. 1986 Sep;83(18):6776–6780. doi: 10.1073/pnas.83.18.6776. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Ohlstein E. H., Storer B., Fujita T., Shebuski R. J. Tissue-type plasminogen activator and streptokinase induce platelet hyperaggregability in the rabbit. Thromb Res. 1987 May 15;46(4):575–585. doi: 10.1016/0049-3848(87)90158-7. [DOI] [PubMed] [Google Scholar]
  24. Pannekoek H., Veerman H., Lambers H., Diergaarde P., Verweij C. L., van Zonneveld A. J., van Mourik J. A. Endothelial plasminogen activator inhibitor (PAI): a new member of the Serpin gene family. EMBO J. 1986 Oct;5(10):2539–2544. doi: 10.1002/j.1460-2075.1986.tb04532.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Rao A. K., Pratt C., Berke A., Jaffe A., Ockene I., Schreiber T. L., Bell W. R., Knatterud G., Robertson T. L., Terrin M. L. Thrombolysis in Myocardial Infarction (TIMI) Trial--phase I: hemorrhagic manifestations and changes in plasma fibrinogen and the fibrinolytic system in patients treated with recombinant tissue plasminogen activator and streptokinase. J Am Coll Cardiol. 1988 Jan;11(1):1–11. doi: 10.1016/0735-1097(88)90158-1. [DOI] [PubMed] [Google Scholar]
  26. Schafer A. I., Adelman B. Plasmin inhibition of platelet function and of arachidonic acid metabolism. J Clin Invest. 1985 Feb;75(2):456–461. doi: 10.1172/JCI111720. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Stricker R. B., Wong D., Shiu D. T., Reyes P. T., Shuman M. A. Activation of plasminogen by tissue plasminogen activator on normal and thrombasthenic platelets: effects on surface proteins and platelet aggregation. Blood. 1986 Jul;68(1):275–280. [PubMed] [Google Scholar]
  28. Verheijen J. H., Chang G. T., Kluft C. Evidence for the occurrence of a fast-acting inhibitor for tissue-type plasminogen activator in human plasma. Thromb Haemost. 1984 Jul 29;51(3):392–395. [PubMed] [Google Scholar]
  29. Verheijen J. H., Mullaart E., Chang G. T., Kluft C., Wijngaards G. A simple, sensitive spectrophotometric assay for extrinsic (tissue-type) plasminogen activator applicable to measurements in plasma. Thromb Haemost. 1982 Dec 27;48(3):266–269. [PubMed] [Google Scholar]
  30. Wiman B., Ljungberg B., Chmielewska J., Urdén G., Blombäck M., Johnsson H. The role of the fibrinolytic system in deep vein thrombosis. J Lab Clin Med. 1985 Feb;105(2):265–270. [PubMed] [Google Scholar]
  31. Wun T. C., Kretzmer K. K. cDNA cloning and expression in E. coli of a plasminogen activator inhibitor (PAI) related to a PAI produced by Hep G2 hepatoma cell. FEBS Lett. 1987 Jan 1;210(1):11–16. doi: 10.1016/0014-5793(87)81288-7. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Clinical Investigation are provided here courtesy of American Society for Clinical Investigation

RESOURCES