Chen Jing, Ding Ling, He Cui, Chen Dan, Deng Su-Rong, Gong Fang, Li Yuan
Central Laboratory,Yongchuan Hospital,Chongqing Medical University,Chongqing 402160,China.
Yongchuan Sub-center,Chongqing Blood Center,Chongqing 402160,China.
Zhongguo Yi Xue Ke Xue Yuan Xue Bao. 2018 Jun 28;40(3):299-307. doi: 10.3881/j.issn.1000-503X.2018.03.001.
Objective To explore the inhibitory effect of aspirin and clopidogrel on platelet adhesion and aggregation behaviors under the physiological flow condition using microfluidic chip technology for health volunteers. Methods Peripheral venous blood samples collected from twelve randomly recruited health volunteers were treated with 20 μmol/L acetylsalicylic acid,50 μmol/L 2-methlthioadenosine-5'-monophosphate triethylammonium salt,and their combination,respectively,with untreated blood samples being control group. The blood samples were flowed through a microchannel modified with type I collagen protein at a physiological relevant shear rate of 1000 s for 300 s,while the fluorescent images of platelet aggregations were dynamic captured using a microscope. Based on the images,the platelet coverage rates were calculated and used as quantitative parameters for evaluating platelet adhesion and aggregation behaviors. Results Under a flow condition of 1000 s shear rate,an expected in vivo-like platelet adhesion and aggregation behaviors were observed at the surfaces of collagen proteins for control blood samples. Aspirin alone or clopidogrel alone suppressed platelet adhesion and aggregation at the later period of flow(200-300 s),while the combination of aspirin and clopidogrel reduced the adhesion numbers of platelets at the earlier stage of flow(≤150 s) and compromised the stability of platelet aggregation at the later period of flow(200-300 s). The combination showed synergistic effect in inhibiting platelet aggregation. Furthermore,such inhibitory effect was heterogeneous among 12 volunteers. Conclusion This simple microfluidic technology can offer a new technical platform for analyzing the inhibitory effect of antiplatelet drugs.
目的 运用微流控芯片技术，探讨阿司匹林和氯吡格雷对健康志愿者在生理流动条件下血小板黏附与聚集行为的抑制作用。方法 从12名随机招募的健康志愿者采集外周静脉血样本，分别用20 μmol/L乙酰水杨酸、50 μmol/L 2 - 甲硫基腺苷 - 5'- 单磷酸三乙铵盐及其组合进行处理，未处理的血样作为对照组。血样以1000 s的生理相关剪切速率流经用I型胶原蛋白修饰的微通道300 s，同时用显微镜动态捕捉血小板聚集的荧光图像。基于这些图像，计算血小板覆盖率，并将其作为评估血小板黏附与聚集行为的定量参数。结果 在1000 s剪切速率的流动条件下，对照组血样在胶原蛋白表面观察到类似体内的血小板黏附与聚集行为。单独使用阿司匹林或氯吡格雷在流动后期（200 - 300 s）抑制血小板黏附与聚集，而阿司匹林和氯吡格雷的组合在流动早期（≤150 s）减少血小板黏附数量，并在流动后期（200 - 300 s）破坏血小板聚集的稳定性。该组合在抑制血小板聚集中显示出协同作用。此外，这种抑制作用在12名志愿者中存在异质性。结论 这种简单的微流控技术可为分析抗血小板药物的抑制作用提供一个新的技术平台。
