Development of a novel method for anticoagulant property of vascular endothlial cells and its application to the diagnosis of cardiovascular diseases

血管内皮细胞的抗凝作用受到多种机制的调节。在这项研究中，我们重点研究了两种蛋白，TFPI和β2-糖蛋白I，这两种蛋白被认为与内皮细胞的抗凝血特性密切相关。TFPI是一种蛋白水解酶抑制剂，具有抑制凝血级联反应的能力。β-糖蛋白I具有与多种负电荷物质结合的能力，在抗磷脂综合征中发挥重要作用。本研究的目的是发展这两种蛋白的新检测方法，并将其应用于临床。我们针对TFPI-Xa的一种复合物制备了特异性的单抗，并建立了一种检测血浆中TFPI-Xa复合物的方法。我们发现DIC患者的这种复合体的水平明显高于正常对照组。我们还制备了针对一种缺口形式的β2-糖蛋白I的特异性单抗，并建立了一种检测血浆中这种缺口形式的方法。我们发现，在DIC和抗心磷脂抗体的患者中，NICK形式的水平显著升高。

The anticoagulant properties of vascular endothelial cells are regulated by various mechanisms. In this study, we focused on two proteins, TFPI and beta2-glycoprotein I, which are supposed to be closely associated with the anicoagulant property of endothelial cells. TFPI is a protease inhibitor with the ability to inhibit the initial reactions of the blood coagulation cascade. Beta-glycoprotein I has the ability to bind with various negatively-charged substances and plays major role in anti-phospholipid syndrome. Our purpose of this study is the development of the novel methods for these two proteins and the clinical application of these methods.We prepared a specific monoclonal antibody for a complex of TFPI-Xa and established a method to measure the complex in plasma. We found that the level of the complex is significantly higher in DIC patients than in normal control.We also prepared a specific monoclonal antibody for a nicked form of beta2-glycoprotein I and established a method to measure the nicked form in plasma. We found that the level of the nicked form is significantly higher in the patients with DIC and anti-cardiolipin antibody.