Huang Xiao-Jing, Zhang Tian-Cong, Gao Xue-Mei, He Cui, Huan Xuan-Rong, Li Yuan
Central Laboratory,Yongchuan Hospital of Chongqing Medical University,Chongqing 402160,China.
Zhongguo Yi Xue Ke Xue Yuan Xue Bao. 2023 Apr;45(2):257-263. doi: 10.3881/j.issn.1000-503X.15155.
Objective To examine the antiplatelet effect of ticagrelor by microfluidic chip and flow cytometry under shear stress . Methods Microfluidic chip was used to examine the effect of ticagrelor on platelet aggregation at the shear rates of 300/s and 1500/s.We adopted the surface coverage of platelet aggregation to calculate the half inhibition rate of ticagrelor.The inhibitory effect of ticagrelor on ADP-induced platelet aggregation was verified by optical turbidimetry.Microfluidic chip was used to construct an vascular stenosis model,with which the platelet reactivity under high shear rate was determined.Furthermore,the effect of ticagrelor on the expression of fibrinogen receptor (PAC-1) and P-selectin (CD62P) on platelet membrane activated by high shear rate was analyzed by flow cytometry. Results At the shear rates of 300/s and 1500/s,ticagrelor inhibited platelet aggregation in a concentration-dependent manner,and the inhibition at 300/s was stronger than that at 1500/s (both <0.001).Ticagrelor at a concentration ≥4 μmol/L almost completely inhibited platelet aggregation.The inhibition of ADP-induced platelet aggregation by ticagrelor was similar to the results under flow conditions and also in a concentration-dependent manner.Ticagrelor inhibited the expression of PAC-1 and CD62P. Conclusion We employed microfluidic chip to analyze platelet aggregation and flow cytometry to detect platelet activation,which can reveal the responses of different patients to ticagrelor.
目的 通过微流控芯片和流式细胞术在剪切应力条件下检测替格瑞洛的抗血小板作用。方法 采用微流控芯片检测替格瑞洛在300/s和1500/s剪切速率下对血小板聚集的影响。采用血小板聚集的表面覆盖率计算替格瑞洛的半抑制率。通过光学比浊法验证替格瑞洛对ADP诱导的血小板聚集的抑制作用。利用微流控芯片构建血管狭窄模型,测定高剪切速率下的血小板反应性。此外,通过流式细胞术分析替格瑞洛对高剪切速率激活的血小板膜上纤维蛋白原受体(PAC-1)和P-选择素(CD62P)表达的影响。结果 在300/s和1500/s剪切速率下,替格瑞洛以浓度依赖性方式抑制血小板聚集,且在300/s时的抑制作用强于1500/s时(均P<0.001)。浓度≥4 μmol/L的替格瑞洛几乎完全抑制血小板聚集。替格瑞洛对ADP诱导的血小板聚集的抑制作用与流动条件下的结果相似,也是浓度依赖性的。替格瑞洛抑制PAC-1和CD62P的表达。结论 我们采用微流控芯片分析血小板聚集,流式细胞术检测血小板活化,可揭示不同患者对替格瑞洛的反应。