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

血管内皮细胞抗凝特性的新方法开发及其在心血管疾病诊断中的应用

• 批准号: 10557095
• 负责人: KATO Hisao
• 金额: $ 2.88万
• 依托单位: National Cardiovascular Center Research Institute
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 1998
• 资助国家: 日本
• 起止时间: 1998 至 1999
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-10557095/
• 关键词: Anticoagulant; endothelial cell; diagnosis; cardiovascular disease; TFPI; beta-2-glycoprotein I; monoclonal antibody; DIC; anti-phospholipid antibody syndrome; 動脈硬化; 測定法

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.

血管内皮细胞的抗凝特性受多种机制调节。在这项研究中，我们集中在两个蛋白质，TFPI和β 2-糖蛋白I，这应该是密切相关的血管内皮细胞的抗凝特性。TFPI是一种蛋白酶抑制剂，能够抑制血液凝固级联的初始反应。β-糖蛋白I具有与多种带负电荷物质结合的能力，在抗磷脂综合征中起主要作用。本研究的目的是建立检测这两种蛋白质的新方法，并探讨其临床应用价值。我们制备了一种特异性抗TFPI-Xa复合物的单克隆抗体，建立了一种检测血浆中TFPI-Xa复合物的方法。我们还制备了β_2-糖蛋白Ⅰ切口型的特异性单克隆抗体，并建立了测定血浆中β_2-糖蛋白Ⅰ切口型的方法。我们发现，在DIC和抗心磷脂抗体的患者中，切口形式的水平显著升高。

项目成果

C.Hine, K.Enjyoji, K.Kokame, S.Nakamura, A.Takei, Y.Kamikubo, K.Sueishi, H.Kato: "Monkey hepatocytes efficiently express tissue factor pathway Inhibitor (TFPI), In contrast with human and rat hepatocytes."J Biochem.. 152. 1039-1047 (1999)

C.Hine、K.Enjyoji、K.Kokame、S.Nakamura、A.Takei、Y.Kamikubo、K.Sueishi、H.Kato：“猴肝细胞有效表达组织因子途径抑制剂 (TFPI)，与人类和

加藤久雄、亀井加恵子: "立体構造からみたTFPI(Tissue Factor Pathway Inhibitor)の作用機序"日本血栓止血学会誌. 6 (1999)

加藤久雄、龟井佳惠子：“从三级结构的角度看 TFPI（组织因子途径抑制剂）的作用机制”日本血栓与止血学会杂志 6（1999 年）。

Chiemi Hine, et al: "Monkey hepatocytes efficiently express tissue factor pathway inhibitor (TFPI), in contrast with human and rat hepatocytes"J. Biochem.. 125(6). 1039-1047 (1999)

Chiemi Hine 等人：“与人类和大鼠肝细胞相比，猴肝细胞有效表达组织因子途径抑制剂 (TFPI)”。

加藤久雄: "血管細胞の抗血栓性機能とTFPI"最新医学社. 6 (2000)

加藤久男：“血管细胞和 TFPI 的抗血栓功能” Shinshin Igakusha 6 (2000)。

N.Ohkura, G.Soe, I. Kohno, K.Kumeda, H.Wada, Y.Kamikubo, H.Shiku, H.Kato: "Monoclonal antibody specific for tissue factor pathway Inhibitor-factor Xa complex: Its characterization and application to plasmas from patients with disseminated Intravascular co

N.Ohkura、G.Soe、I. Kohno、K.Kumeda、H.Wada、Y.Kamikubo、H.Shiku、H.Kato：“组织因子途径抑制剂-因子 Xa 复合物特异性单克隆抗体：其表征和应用