Development of mechanical device to continuously guantitute platelet thrunbus formation under flow conditions.

开发在流动条件下连续定量血小板血栓形成的机械装置。

• 批准号: 07557079
• 负责人: IKEDA Yasuo
• 金额: $ 8.64万
• 依托单位: Keio University School of Medicine
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (A)
• 财政年份: 1995
• 资助国家: 日本
• 起止时间: 1995 至 1997
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07557079/
• 关键词: Platelet adhesion; Wall shear rate; Hematocrit; Platelet membrane glycoproteins; Adhesion molecules; Anti-platelet agents; ずり速度; 血栓

Platelet adhesion to subendothelium, one of the earliest steps in primary hemostasis and pathological thrombus formation is followed by subsequent platelet plug formation including platelet-platelet interactions. Therefore, it is difficult to seggregate from the subsequent platelet interactions and analyze the mechanism of platelet adhesion only. We developed a recirculating flow system including a quartz crystal microbalance (QCM) as a sensor which allows continuous monitoring of the surface coverage of firmly adherent platelets only. Using this system, we found that platelet adhesions to collagen (bovine tendon type 1 : CL) and human von Willebrand factor (vWf) were dependent upon wall shear rate (310/s-2,400/s) and hematocrit (20-45%). It was found that there existed two recptor-ligand pairs i.e. platelet glycoprotein (GP) Ib/IX-vWf and GPIa/IIa-CL independently operating in platelet adhesion to CL evenunder higher flow conditions. Interestingly, well-known anti-platelet agents such as aspirin and cilostazol were found to enhance platelet adhesion to CL,although they readily inhibited subsequent platelet-platelet interactions. Thus, our newly-developed QCM system may be useful to study the mechanism of platelet adhesion under flow conditions and to screen the efficacy of new anti-platelet adhesion drugs.

血小板粘附于下皮下，这是原发性止血和病理血栓形成的最早步骤之一，然后是随后的血小板塞形成，包括血小板 - 血小板相互作用。因此，很难从随后的血小板相互作用中分离并仅分析血小板粘附的机理。我们开发了一个循环流系统，包括一个石英晶体微平衡（QCM）作为传感器，该传感器只能连续监视仅牢固粘附血小板的表面覆盖范围。使用该系统，我们发现对胶原蛋白（牛肌腱1：Cl）和人von Willebrand因子（VWF）的血小板粘附取决于壁剪切速率（310/S-2,400/s）和血细胞比容（20-45％）。发现存在两个循环 - 配体对，即血小板糖蛋白（GP）IB/IX-VWF和GPIA/IIA-CL独立于血小板粘附起作用，可与CL相关。有趣的是，尽管它们很容易抑制随后的血小板 - 血小板相互作用，但发现了众所周知的抗血域剂，例如阿司匹林和西洛唑嗪可增强血小板的粘附。因此，我们新开发的QCM系统可能有助于研究流动条件下血小板粘附的机理，并筛选新的抗血域粘附药的功效。

项目成果

Kameda H, Morita I, Handa M., Ikeda Yetal: "Re-expression of functional P-selectin molecules on the endothelial cell surface by repeated stimulation with thrombin." Brit J Haematol 97. 97. 348-355 (1997.)

Kameda H、Morita I、Handa M.、Ikeda Yetal：“通过凝血酶的重复刺激在内皮细胞表面重新表达功能性 P-选择素分子。”

Kitaguchi T: "Establishment and characterization of transgenic mice expressing human platelet glycoprotein lba" 220. 418-424 (1996)

Kitaguchi T：“表达人血小板糖蛋白 lba 的转基因小鼠的建立和表征”220. 418-424 (1996)

Yokoyama K.,Handa M.,and Ideda Y.et al: "Characterization of the novel murine monoclonal anti-von Willebrand factor (vWf) antibody GUR76-23 which inhibits vWf interaction with α IIb β 3 but not α v β 3 integrin." Biochem Biophys Res Commun. 234. 147-152 (

Yokoyama K.、Handa M. 和 Ideda Y.等人：“新型鼠单克隆抗血管性血友病因子 (vWf) 抗体 GUR76-23 的表征，该抗体抑制 vWf 与 α IIb β 3 但不与 α v β 3 整合素相互作用“Biochem Biophys Res Commun.234.147-152（

Goto S.,Sakai H.,Ikeda Y,and Handa S.: "Acute myocardial infarction plasma augments platelet thrombus growth in high shear rates." The Lancet. 349(9051). 543-544 (1997)

Goto S.、Sakai H.、Ikeda Y 和 Handa S.：“急性心肌梗死血浆在高剪切速率下会增强血小板血栓的生长。”

Iijima K, Murata M., Handa M., Ikeda Y.et al: "High shear stress attenuates agonist-induced,glycoprotein IIb/IIIa-mediated platelet aggregation when von Willebrand factor binding to glycoprotein Ib/IX is blocked." Biochem Biophys Res Commun. 233(3). 796-8

Iijima K、Murata M.、Handa M.、Ikeda Y.等人：“当冯维勒布兰德因子与糖蛋白 Ib/IX 结合被阻断时，高剪切应力会减弱激动剂诱导的糖蛋白 IIb/IIIa 介导的血小板聚集。”