Stuies on the Mechanisms of Thrombus-formation and Thrombolysis with Special Referencies to Clinical Diagnosis, Treatment and Prophyraxis of Thrombosis

本课题旨在阐明血栓形成的重要因素，即内皮细胞功能的细胞生物学特性、内皮细胞损伤的危险因素、血小板黏附和聚集的分子机制、凝血-纤溶系统启动和调节因素的生化和免疫组织化学研究。此外，在此基础上，对凝血和纤溶功能的临床或临床前变化进行了酶学和免疫化学分析，检测了血小板和凝血-纤溶系统的几个因素。我们建立了几种丝氨酸蛋白酶及其抑制物的免疫酶分析方法和特异性酶分析方法。血栓形成和溶栓机制的研究：内皮细胞合成并表达抗血栓形成因子或功能。结果：1.分析了凝血因子VII、血栓调节蛋白_2PI、蛋白C、S蛋白、纤溶酶原激活剂抑制物(1型)、酸性胰蛋白酶抑制物和钙结合蛋白的免疫化学性质和/或氨基酸序列。临床血栓形成的发病机制、诊断和治疗的研究：以胶体金为示踪剂，对糖蛋白I、IIb/IIIa的定位进行了免疫结构研究。这些膜蛋白参与了血小板的黏附和聚集，与凝血酶和纤维蛋白原结合有关。用MCAS建立了检测血小板环氧合酶和12-脂氧合酶的酶联免疫吸附试验系统。口服和静脉注射尿激酶对脑血管血栓形成和急性心肌梗死有效。

This project was co-operatively performed to elucidate the recent topics as the important factors participating in the thrombosis, namely cell-biological characteristics of endothelial cell function, risk factors of endothelial injuries, the molecular mechanism of platelet adherence and aggregation, biochemical and immunohistochemical studies of initiating and regulating factors of coagulation-fibrinolysis system. In addition, on these basic researches clinical or preclinical alterations of coagulatory and fibrinolytic functions were enzymologically and immunochemically analyzed for platelets and several factors of coagulation-fibrinolysis system. We established the immunoenzymatic and specific enzymatic assay methods for several serine-proteases and inhibitors.1. Studies on the mechanism of thrombus-formation and thrombolysis: Endothelial cells synthesized and expressed not only the antithrombogenic but also thrombogenic factors or functions. Factor VII, thrombomodulin, _2PI, protein C, protein S,PA inhibitor (type 1), acid-stable trypsin inhibitor and calphobindin were analyzed for immunochemical properties and/or amino acid and cDNA sequences.2. Studies on the pathogenesis, diagnosis and treatment of clinical thrombosis: The localization of glycoprotein I, IIb/IIIa was immunoultrastructurally investigated using collodal gold as a tracer. These membrane proteins were participated in platelet adherence and aggregation, associated with the thrombin and fibrinogen binding. ELISA systems for measuring cycloxygenase and 12-lipoxygenase in platelets were developed using MCAs. Oral and intravenous administrations of urokinase were effective for cerebrovascular thrombosis and acute myocardial infarction.