CYP2C19基因多态性影响稳定型冠心病患者氯吡格雷治疗后血小板P2Y_(12)受体信号通路的分子机制

Effect of CYP2C19 polymorphism on platelet P2Y_(12) receptor signaling pathway in Chinese patients with stable coronary artery disease after the treatment of clopidogrel

目的探讨中国冠心病患者CYP2C19基因多态性影响氯吡格雷疗效的分子机制。方法入组208例接受冠状动脉介入治疗(percutaneous coronary intervention,PCI)的稳定型冠心病(stable coronary artery disease,SCAD)患者,采用PCR产物直接测序的方法对所有纳入对象进行CYP2C19基因多态性分析,血栓弹力图检测300 mg氯吡格雷负荷量24 h后的血小板聚集率,流式细胞仪检测P2Y_(12)受体下游信号分子血管扩张刺激磷蛋白血小板反应性指数(vasodilator-stimulated phosphorylation-platelet reactivity index,VASP-PRI)和Akt磷酸化(P-Akt)水平。结果 208例患者中85例(40.9%)的基因型为CYP2C19*1/*1(非携带者),无等位基因缺失(loss-of-function,LOF),93例(44.7%)为CYP2C19*1/*2或CYP2C19*1/*3(1个LOF),30例(14.4%)为CYP2C19*2/*2或CYP2C19*2/*3(2个LOF)。氯吡格雷治疗后的血小板聚集率及VASPPRI随着携带LOF数目的增加而增加,而P-Akt只在2个LOF的情况下才会明显增加。VASP-PRI与氯吡格雷治疗后血小板聚集率有相关性(r=0.672,P<0.001),但是与P-Akt不相关。结论携带2个CYP2C19 LOF的SCAD患者经氯吡格雷治疗后血小板聚集率更高,并与活性更高的G蛋白偶联受体P2Y_(12)信号通路相关。

Objective To investigate the molecular mechanism of the effect of CYP2C19 polymorphism on the efficacy of clopidogrel in patients with stable coronary artery disease （SCAD）. Methods A total of 208 patients with coronary artery disease undergoing percutaneous coronary intervention （PCI） were included. The CYP2C19 variant alleles were detected by gene sequencing. Platelet reactivity was assessed by thrombelastograph at 24 h after 300 mg clopidogrel loading. P2Y12-Gi signaling was measured by flow cytometric analysis of vasodilator-stimulated phosphorylation-platelet reactivity index （VASP-PRI） and Akt phosphorylation （P-Akt） index. Results Among 208 patients,40.9% were classified as CYP2C19＊1/＊1 （non-carriers） with no loss-of-function （LOF）, 44.7% as CYP2C19＊1/＊2 or CYP2C19＊1/＊3 （one LOF carriers） and 14.4% as CYP2C19＊2/＊2 or CYP2C19＊2/＊3 （two LOF carriers）. Both postclopidogrel platelet aggregation and VASP-PRI increased by the presence of one LOF allele, and were even more pronounced with the presence of two LOF alleles. In contrast, P-Akt index only increased in two LOF carriers. VASP-PRI was positively associated with postclopidogrel platelet aggregation （r=0.661, P〈0.001）, but not with P-Akt index. Conclusions The two CYP2C19 LOF carriage was associated with more active state of P2Y12-Gi signaling in postclopidogrel platelet in Chinese patients with SCAD.