Lack of tPA Significantly Affects Antithrombotic Therapy by a GPIIb/IIIa Antagonist, but not by a Thrombin Inhibitor in Mice

Summary

The interaction of fibrinolytic components with platelets or coagulation factors after endothelial injury, was investigated in mouse deficient in tissue type plasminogen activator (tPA –/–), or urokinase (uPA –/–) and in their wild type control (tPA +/+, uPA +/+). A thrombus was induced in the murine carotid artery using the photochemical reaction. Blood flow was continuously monitored and the time needed before the vessel became completely obstructed was within 11 min in all types of mice. When GR144053, a platelet glycoprotein IIb/IIIa antagonist, or argatroban, a thrombin inhibitor, was applied, the time required to occlusion was prolonged in a dose-dependent manner in all types of mice. However, when GR144053 was injected in tPA –/– mice, the most significant changes were observed: that is the estimated ED50 was 14.8 times higher than the one in tPA +/+ mice. On the other hand, when argatroban was injected in tPA –/– mice, the estimated ED50 was not changed. Platelet aggregation, haemostasis tests and bleeding times were not significantly different among the different types of mice. In conclusion, the antithrombotic effect of platelet inhibition by a GPIIb/IIIa antagonist, is severely affected by the absence or presence of tPA-production. Thus, the lack of tPA significantly reduces the antithrombotic efficacy.

Keywords

GPIIb/IIIa - platelets - thrombus - tPA - thrombin

• References

• 1 Fuster V. Mechanisms leading to myocardial infarction: insights from studies of vascular biology. Circulation 1994; 90: 2126-46.
• 2 Haines ST, Bussey HI. Thrombosis and the pharmacology of antithrombotic agents. Ann of Pharmacother 1995; 29: 892-905.
• 3 Coller BS, Folts JD, Scudder LE, Smith SR. Antithrombotic effect of a monoclonal antibody to the platelet glycoprotein IIb/IIIa receptor in an experimental animal model. Blood 1986; 68: 783-6.
• 4 Kiss RG, Stassen JM, Deckmyn H, Roskam T, Gold HK, Plow EF, Collen D. Contribution of platelets and the vessel wall to the antithrombotic effects of a single bolus injection of Fab fragments of the antiplatelet GPIIb/IIIa antibody 7E3 in a canine arterial eversion graft preparation. Arteriosclerosis & Thrombosis 1994; 14: 375-80.
• 5 Konstantopoulos K, Kamat SG, Schafer AI, Banez EI, Jordan R, Kleiman NS, Hellums JD. Shear-induced platelet aggregation is inhibited by in vivo infusion of an anti-glycoprotein IIb/IIIa antibody fragment, c7E3 Fab, in patients undergoing coronary angioplasty. Circulation 1995; 91: 1427-31.
• 6 Lunven C, Gauffeny C, Lecoffre C, O’Brien DP, Roome NO, Berry CN. Inhibition by Argatroban, a specific thrombin inhibitor, of platelet activation by fibrin clot-associated thrombin. Thromb Haemost 1996; 75: 154-60.
• 7 Catella-Lawson F. Direct thrombin inhibitors in cardiovascular disease. Coronary Artery Disease 1997; 08: 105-11.
• 8 Matsuo O, Bando H, Okada K, Tanaka K, Tsukada M, Iga Y, Arimura H. Thrombolytic effect of single-chain pro-urokinase in a rabbit jugular vein. Thrombos Res 1986; 42: 187-94.
• 9 Collen D, Lijnen HR. Basic and clinical aspects of fibrinolysis and thrombolysis. Blood 1991; 78: 3114-24.
• 10 Plow EF, Herren T, Redlitz A, MIles LA, Hoover-Plow JL. The cell biology of the plasminogen system. FASEB J 1995; 09: 939-45.
• 11 Levi M, Biemond BJ, Van Zonneveld AJ, Ten Cate JW, Pannekoek H. Inhibition of plasminogen activator inhibitor-1 activity results in promotion of endogenous thrombolysis and inhibition of thrombus extension in models of experimental thrombosis. Circulation 1992; 85: 305-12.
• 12 Matsuno H, Uematsu T, Nagashima S, Nakashima M. Photochemically induced thrombosis model in rat femoral artery and evaluation of effects of heparin and tissue-type plasminogen activator with use of this model. J Pharmacol Methods 1991; 25: 303-317.
• 13 Matsuno H, Uematsu T, Umemura K, Takiguchi Y, Wada K, Nakashima M. Effects of vapiprost, a novel thromboxane receptor antagonist, on thrombus formation and vascular patency after thrombolysis by tissue-type plasminogen activator. B J Pharmacol 1992; 106: 533-8.
• 14 Matsuno H, Uematsu T, Nakashima M. Thrombolytic effect of a plasminogen-plasminogen activator chimera in a photochemically induced thrombosis (PIT) model. B J Pharmacol 1993; 110: 1278-9.
• 15 Hirata Y, Takiguchi Y, Wada K, Matsuno H, Umemura K, Uematsu T, Nakashima M. Roles of platelet-activating factor, thromboxane A2, ADP and thrombin in thrombogenesis in the guinea pig. Eur J Pharmacol 1993; 231: 421-5.
• 16 Matsuno H, Uematsu T, Kozawa O, Niwa M, Ueshima S, Matsuo O. Fibrinolytic factors play a different role on the development or lysis of thrombus formation. Thromb Haemost 1999; 81: 601-4.
• 17 Kawasaki T, Sakai Y, Taniuchi Y, Sato K, Maruyama K, Shimizu M, Kaku S, Yano S, Inagaki O, Tomioka K, Yanagisawa I, Takenaka T. Biochemical characterization of a new disintegrin, flavostatin, isolated from Trimeresurus flavoviridis venom. Biochimie 1996; 78: 245-52.
• 18 Okamoto S, Hijikata A, Kikumoto R, Tonomura S, Hara H, Ninomiya K, Maruyama A, Sugano M, Tamao Y. Potent inhibition of thrombin by the newly synthesized arginine derivative No. 805. The importance of stereo-structure of its hydrophobic carboxamide portion. BBRC 1981; 101: 440-6.
• 19 Carmeliet P, Schoonjans L, Kieckens L, Ream B, Degen J, Bronson R, De Vos R, van den Oord JJ, Collen D, Mulligan R. Physiological consequences of loss of plasminogen activator gene function in mice. Nature 1994; 368: 419-24.
• 20 Carmeliet P, Stassen JM, Schoonjans L, Ream B, van den Oord JJ, De Mol M, Mulligan RC, Collen D. Plasminogen activator inhibitor-1 gene-deficient mice. II. Effects on hemostasis, thrombosis, and thrombolysis. J Clin Invest 1993; 92: 2756-60.
• 21 Matsuno H, Stassen JM, Vermylen J, Deckmyn H. Inhibition of integrin function by a cyclic RGD-containing peptide. Circulation 1994; 90: 2203-6.
• 22 Mousa SA, DeGrado WF, Mu DX, Kapil RP, Lucchesi BR, Reilly TM. Oral antiplatelet, antithrombotic efficacy of DMP 728, a novel platelet GPIIb/IIIa antagonist. Circulation 1996; 93: 537-43.
• 23 Matsuno H, Kozawa O, Niwa M, Ito T, Tanabe K, Nishida M, Hayashi H, Uematsu T. Effect of GR144053, a fibrinogen-receptor antagonist, on thrombus formation and vascular patency after thrombolysis by tPA in the injured carotid artery of the hamster. J Cardiovasc Pharmacol 1998; 32: 191-7.
• 24 Frederick LG, Suleymanov OD, King LW, SALYERS AK, Nicholson NS, Feigen LP. The protective dose of the potent GPIIb/IIIa antagonist SC-54701A is reduced when used in combination with aspirin and heparin in a canine model of coronary artery thrombosis. Circulation 1996; 93: 129-34.
• 25 Ito T, Matsuno H, Kozawa O, Niwa M, Sakai N, Uematsu T. Comparison of the antithrombotic effects and bleeding risk of fractionated ATA and GPIIb/IIIa antagonist, GR144053 in a hamster model of stenosis. Thromb Res 1999; 95: 49-61.
• 26 Kamat SG, Michelson AD, Benoit SE, Moake JL, Rajasekhar D, Hellums JD, Kroll MH, Schafer AI. Fibrinolysis inhibits shear stress-induced platelet aggregation. Circulation 1995; 92: 1399-1407.
• 27 Strickland S, Reich E, Sherman MI. Plasminogen activator in early embryogenesis: enzyme production by trophoblast and parietal endoderm. Cell 1976; 09: 231-40.
• 28 Rifkin DB, Moscatelli D, Bizik J, Quarto N, Blei F, Dennis P, Flaumenhaft R, Mignatti P. Growth factor control of extracellular proteolysis. Cell Different Develop 1990; 32: 313-8.
• 29 Zhang J, Ren S, Sun D, Shen GX. Influence of glycation on LDL-induced generation of fibrinolytic regulators in vascular endothelial cells. Arteriosclerosis, Thrombosis Vasc Biol 1998; 18: 1140-8.
• 30 Terres W, Lund GK, Hubner A, Ehlert A, Reuter H, Hamm CW. Endogenous tissue plasminogen activator and platelet reactivity as risk factors for reocclusion after recanalization of chronic total coronary occlusions. Am Heart J 1995; 130: 711-6.