Flow cytometric analysis of intraplatelet VASP phosphorylation for the detection of clopidogrel resistance in patients with ischemic cardiovascular diseases
Flow cytometric analysis of intraplatelet VASP phosphorylation for the detection of clopidogrel resistance in patients with ischemic cardiovascular diseases
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DOI：
10.1111/j.1538-7836.2004.01063.x
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发表时间：
2005-01-01
影响因子：
10.4
通讯作者：
Gachet, C
中科院分区：
文献类型：
--
作者：
Aleil, B;Ravanat, C;Gachet, C
在一些缺血性心血管疾病患者中，氯吡格雷对血小板功能的抑制作用存在个体差异，导致氯吡格雷抵抗。因此，需要一个可靠的实验室检查，以确定这种抗血小板治疗保护不足的患者。血管舒张刺激磷蛋白（VASP）是一种血小板内肌动蛋白调节蛋白，其磷酸化依赖于血小板P2 Y（12）受体的活化水平，而血小板P2 Y（12）受体是氯吡格雷的靶点。本研究的目的是使用流式细胞术VASP磷酸化测定来评估氯吡格雷治疗的疗效。血小板反应性指数（PRI）以百分比表示，是静息（+PGE（1））和活化（+ADP）血小板之间VASP荧光强度的差异。在体外实验中，PRI与竞争性拮抗剂AR-C69931 MX特异性阻断P2 Y（12）受体诱导的血小板聚集抑制作用呈强相关（R = 0.72，P < 0.0001）。离体实验中，47名健康供体的PRI为78.3 +/- 4.6%，34名未接受氯吡格雷治疗的患者为79.0 +/- 4.1%，33名接受氯吡格雷治疗的患者为61.1 +/- 17.0%（P < 0.0001）。在氯吡格雷组中，PRI值分布广泛（从6.6%到85.8%），超过30%的患者的PRI值相当于未接受氯吡格雷的患者。VASP磷酸化的流式细胞术分析似乎是一个合适的测试，以评估氯吡格雷治疗的疗效。该试验表明，血小板对氯吡格雷的抑制反应存在广泛的个体间差异，并显示三分之一的治疗患者似乎对该疗法“无保护”。
Interindividual variability of the inhibitory effect of clopidogrel on platelet functions leading to clopidogrel resistance has been described in some patients with ischemic cardiovascular disease. A reliable laboratory test is therefore needed to identify patients insufficiently protected by this antiplatelet treatment. The phosphorylation of vasodilator-stimulated phosphoprotein (VASP), an intraplatelet actin regulatory protein, is dependent on the level of activation of the platelet P2Y(12) receptor, which is targeted by clopidogrel. The aim of this study was to use a flow cytometric VASP phosphorylation assay to evaluate the efficacy of clopidogrel therapy. The platelet reactivity index (PRI), expressed as a percentage, is the difference in VASP fluorescence intensity between resting (+PGE(1)) and activated (+ADP) platelets. In vitro, the PRI was strongly correlated with the inhibition of platelet aggregation induced by specific blockade of the P2Y(12) receptor by the competitive antagonist AR-C69931MX (R = 0.72, P < 0.0001). Ex vivo, the PRI was 78.3 +/- 4.6% in 47 healthy donors, 79.0 +/- 4.1% in 34 patients not receiving clopidogrel and 61.1 +/- 17.0% in 33 patients treated with clopidogrel (P < 0.0001). In the clopidogrel group, the PRI values were widely dispersed (from 6.6 to 85.8%) and more than 30% of these patients had a PRI equivalent of values in patients not receiving clopidogrel. The flow cytometric analysis of VASP phosphorylation seems to be a suitable test to evaluate the efficacy of clopidogrel treatment. This assay demonstrated a wide interindividual variability of the inhibitory response of platelets to clopidogrel and showed that one-third of the patients treated appeared to be 'unprotected' by this therapy.