Function of Tissue-type Plasminogen Activator Releaser on Vascular Endothelial Cells and Thrombolysis In Vivo

Summary

The effect of monosodium[2-(6-hydroxynaphthalen-2-yl)-6-methylpyrimidin-4-yloxy]acetate dihydrate (JTV-926) on fibrinolysis was investigated in vitro and in vivo. JTV-926 released tissue-type plasminogen activator (t-PA) from human vascular endothelial cells in a dose-dependent manner. The thrombolytic effect of JTV-926 was studied using three animal thrombosis models; a photo-irradiation-induced mouse carotid artery thrombosis model, a photo-irradiation-induced rat femoral artery thrombosis model and a thrombin-induced rat venous thrombosis model. In the mouse thrombosis model, t-PA deficient mice (t-PA−/−mice) and their wild-type (t-PA+/+) were used. JTV-926 was injected as a bolus 30 min after the interruption of blood flow by an occlusion thrombi. Blood flow was continuously monitored for 180 min after intravenous administration of JTV-926 (1 mg/kg). Although the recanalization rate of the occluded artery was 37.5% in t-PA +/+ mice with the vehicle control, it increased to 75% in t-PA+/+ mice after JTV-926 administration. However, when JTV-926 was administrated in t-PA−/−mice, vascular recanalization was not observed in any arteries. In the photo-irradiation-induced rat femoral artery thrombosis model, intra-duodenal administration of JTV-926 induced thrombolysis. Moreover, in the thrombin-induced rat venous thrombosis model, the dose-dependent thrombolysis was also observed by oral administration of JTV-926. It was suggested that JTV-926 revealed a sufficient thrombolytic effect through the absorption from the intestine. Thus, a newly synthesized compound, JTV-926 induced t-PA release from vascular endothelial cells and effective thrombolysis in vivo.

Keywords

Endothelial cells - t-PA - thrombolysis - t-PA deficient mice - animal model

• References

• 1 ISIS-2 (Second International Study of Infarct Survival) Collaborative Group. Randomised trial of intravenous streptokinase, oral aspirin, both or neither among 17,187 cases of suspected acute myocardial infarction: ISIS-2. Lancet 1988; 02: 349-60.
• 2 Matsuo O, Mihara H, Rokushima Y. Assessment of the thrombolytic effect of low urokinase concentrations. Acta Hematol Jap 1976; 39: 298-305.
• 3 Matsuo O, Rijken DC, Collen D. Thrombolysis by human tissue plasminogen activator and urokinase in rabbits with experimental pulmonary embolus. Nature 1981; 291: 590-1.
• 4 Bando H, Okada K, Matsuo O. Thrombolytic effect of pro-urokinase in vivo . Fibrinolysis 1987; 01: 169-76.
• 5 Smith RAG, Dupe RJ, English PD, Green J. Fibrinolysis with acyl-enzymes: A new approach to thrombolytic therapy. Nature 1981; 290: 505-8.
• 6 Matsuo O, Okada K, Fukao H, Tomioka Y, Ueshima S, Watanuki M, Sakai M. Thrombolytic properties of staphylokinase. Blood 1990; 76: 925-9.
• 7 Collen D, Lijnen HR. Basic and clinical aspects of fibrinolysis and thrombolysis. Blood 1991; 78: 3114-24.
• 8 Korninger C, Stassen JM, Collen D. Turnover of human extrinsic (tissue-type) plasminogen activator in rabbits. Thromb Haemost 1984; 46: 658-61.
• 9 Huber K, Kirchheimer JC, Korninger C, Binder BR. Hepatic synthesis and clearance of components of the fibrinolytic system in healthy volunteers and in patients with different stages of liver cirrhosis. Thromb Res 1991; 62: 491-500.
• 10 Gulba DC, Bode C, Runge MS, Huber K. Thrombolytic agents – an overview. Ann Hematol 1996; 73: S9-S27.
• 11 Collen D, Lijnen HR. Recent developments in thrombolytic therapy. Fibrinolysis Proteol 2000; 14: 66-72.
• 12 Levin EG, Loskutoff DJ. Cultured bovine endothelial cells produce both urokinase and tissue-type plasminogen activators. J Cell Biol 1982; 94: 631-6.
• 13 Rijken DC, van Hinsbergh VW, Sens EH. Quantitation of tissue-type plasminogen activator in human endothelial cell cultures by use of an enzyme immunoassay. Thromb Res 1984; 33: 145-53.
• 14 Petaja J. Fibrinolytic response to venous occlusion for 10 and 20 minutes in healthy subjects and in patients with deep vein thrombosis. Thromb Res 1989; 56: 251-63.
• 15 Grader AMA, da Costa J, Cash JD. A new vasopressin analogue and fibrinolysis. Lancet 1973; 02: 1417-8.
• 16 Butler WB, Kirkland WL, Gargala TL, Goran N, Kelsey WH, Berlinski PJ. Steroid stimulation of plasminogen activator production in a human breast cancer cell line (MCF-7). Cancer Res 1983; 43: 1637-41.
• 17 Hagiwara H, Shimonaka M, Morisaki M, Ikekawa N, Inada Y. Sitosterolstimulative production of plasminogen activator in cultured endothelial cells from bovine carotid artery. Thromb Res 1984; 33: 363-70.
• 18 Okayama T, Nakano M, Odake S, Hagiwara M, Morikawa T, Ueshima S. et al. Synthetic dipeptide, N-stearoyl-D-Ser-L-Pro-OEt, induces release of tissue-type plasminogen activator in cultured cells and in experimental animals. Chem Pharm Bull 1994; 42: 1854-8.
• 19 Carmeliet P, Schoonjans L, Kiecjens L, Ream B, Degen J, Bronson R, DeVos R, van den Oord JJ, Collen D. Physiological consequence of loss of plasminogen activator gene function in mice. Nature 1994; 368: 419-24.
• 20 Matsumoto H, Ueshima S, Fukao H, Mitsui Y, Matsuo O. Identification of urokinase-type plasminogen activator receptor in human endothelial cells and its modulation by phorbol myristate acetate. Cell Struct Funct 1995; 20: 429-37.
• 21 Ueshima S, Fukao H, Okada K, Matsuo O. The effect of argatroban on injured endothelial cells by thrombin. Blood Coagul Fibrin 2000; 11: 631-9.
• 22 Fukao H, Hagiya Y, Ueshima S, Okada K, Takahashi T, Izaki S, Matsuo O. Effects of heat shock on the expression of urokinase-type plasminogen activator receptor in human umbilical vein endothelial cells. Thromb Haemost 1996; 75: 352-8.
• 23 Matsuo O, Sakai T, Bando H, Okada K, Nakajima S, Takagi O, Izaki S. Plasminogen activator in broncho-alveolar fluid. Haemostas 1986; 16: 43-50.
• 24 Matsuno H, Kozawa O, Niwa M, Ueshima S, Matsuo O, Collen D, Uematsu T. Differential role of components of the fibrinolytic system in the formation and removed of thrombus induced by endothelial injury. Thromb Haemost 1999; 81: 601-4.
• 25 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 Meth 1991; 25: 303-17.