Comparative real-time effects on platelet adhesion and aggregation under flowing conditions of in vivo aspirin, heparin, and monoclonal antibody fragment against glycoprotein IIb-IIIa.
Comparative real-time effects on platelet adhesion and aggregation under flowing conditions of in vivo aspirin, heparin, and monoclonal antibody fragment against glycoprotein IIb-IIIa.
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体内阿司匹林、肝素和抗糖蛋白 IIb-IIIa 单克隆抗体片段在流动条件下对血小板粘附和聚集的实时影响比较。
DOI：
10.1161/01.cir.91.5.1354
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发表时间：
1995
期刊：
影响因子：
37.8
通讯作者：
McIntire,LV
中科院分区：
文献类型：
--
作者：
Turner,NA;Moake,JL;Kamat,SG;Schafer,AI;Kleiman,NS;Jordan,R;McIntire,LV
背景在类似于血管成形术在体内产生的动脉流动条件下，人血小板血栓形成的实时体外系统将有助于评价与体内试验相关的潜在抗动脉血栓形成剂。阿司匹林、肝素和针对血小板糖蛋白（GP）IIb-IIIa的嵌合单克隆抗体抗原结合片段7E3（c7E3 Fab）已被用于尝试延迟或预防血管成形术后冠状动脉的血栓性再闭塞。我们比较了这些药物在体内对GPIb介导的血小板与血管性血友病因子（vWF）/I型胶原粘附的影响。（如在动脉粥样硬化内皮下）和随后的GPIIb-IIIa-纤维蛋白原/vWF介导的血小板聚集在流动条件下类似于那些在收缩的冠状动脉。将来自患者和健康供体的标记血小板在37 ℃下以1500秒-1的异常升高的剪切速率（动脉壁剪切应力为50 - 60达因/cm 2）在胶原I/vWF上灌注1分钟。粘附的荧光血小板的数量每15秒用低亮度摄像机和落射荧光显微镜定量。在5名健康供体摄入975 mg阿司匹林后，与所有实验中的对照血液相比，阿司匹林处理血液中的血小板粘附不受影响（10/10），并且随后的聚集在大多数运行中没有变化（8/10）。在注射12 000 U肝素之前和之后以及输注0.25 mg/kg c7E3 Fab后2分钟、2小时和24小时，分析3名接受血管成形术的阿司匹林治疗患者的血液。在这些患者中，大剂量肝素不能抑制血小板与胶原I/vWF的粘附或随后的聚集。与此相反，在所有3名患者中，输注c7E3 Fab后2分钟，血小板聚集抑制> 50%，并且在c7E3 Fab后2小时和24小时，3名患者中的2名持续抑制。或血小板GPIIb-IIIa的某些其它配体。血小板粘附胶原蛋白I/vWF不受影响。本研究描述了一种动脉损伤的体外模型（类似于血管成形术），该模型使用人血直接比较阿司匹林、肝素和c7 E3 Fab对血小板粘附和随后的聚集的精确相对影响，这是真实的时间。
BackgroundA real-time in vitro system of human platelet thrombosis under arterylike flowing conditions similar to those produced in vivo by angioplasty would be useful for the evaluation of potential antiarterial thrombotic agents in association with in vivo trials. Aspirin, heparin, and the chimeric monoclonal antibody antigen-binding fragment 7E3 (c7E3 Fab) directed against platelet glycoprotein (GP) IIb-IIIa have been used in attempts to delay or prevent thrombotic reocclusion of coronary arteries after angioplasty. We compared the effects of these agents administered in vivo on GPIb-mediated platelet adhesion to von Willebrand factor (vWF)/collagen type I (as in atherosclerotic subendothelium) and on subsequent GPIIb-IIIa–fibrinogen/vWF–mediated platelet aggregation under flowing conditions analogous to those in constricted coronary arteries.Methods and ResultsCitrated whole blood containing mepacrine-labeled platelets from patients and healthy donors was perfused for 1 minute at an abnormally elevated shear rate of 1500 seconds−1(arterial wall shear stress of 50 to 60 dynes/cm2) at 37°C over collagen I/vWF. The number of adherent fluorescent platelets was quantified every 15 seconds with a low-light-level video camera and epifluorescent microscopy. After 5 healthy donors had ingested 975 mg aspirin, platelet adhesion was unaffected in the aspirin-treated blood compared with the control blood in all experiments (10 of 10), and subsequent aggregation was unchanged in most runs (8 of 10). The blood of 3 aspirin-treated patients undergoing angioplasty was analyzed before and after a 12 000-U heparin injection and 2 minutes, 2 hours, and 24 hours after infusion of 0.25 mg/kg of c7E3 Fab. In these patients, the bolus of heparin did not inhibit either platelet adhesion to collagen I/vWF or subsequent aggregation. In contrast, there was >50% inhibition of platelet aggregation 2 minutes after the infusion of c7E3 Fab in all 3 patients, and inhibition persisted in 2 of the 3 patients at 2 hours and 24 hours after c7E3 Fab.ConclusionsIn contrast to aspirin or heparin, the in vivo injection of c7E3 Fab considerably reduces platelet aggregate formation mediated by the binding of fibrinogen, vWF, or some other ligand to platelet GPIIb-IIIa under conditions of abnormally increased shear stress analogous to those in narrowed coronary arteries. Platelet adherence to collagen I/vWF is not affected. This study describes an in vitro model of arterial injury (similar to angioplasty) that uses human blood to compare directly, in real time, the precise relative effects of aspirin, heparin, and c7E3 Fab on platelet adhesion and subsequent aggregation.