血小板 miR-223 表达降低与氯吡格雷抵抗的血小板高反应性相关。

Decreased platelet miR-223 expression is associated with high on-clopidogrel platelet reactivity.

机构信息

Institute of Cardiovascular Disease and Heart Center, Pingjin Hospital, Logistics University of the Chinese People's Armed Police Forces, China.

出版信息

Thromb Res. 2013 Jun;131(6):508-13. doi: 10.1016/j.thromres.2013.02.015. Epub 2013 Mar 15.

DOI:10.1016/j.thromres.2013.02.015

PMID:23498169

Abstract

OBJECTIVES

We aimed to investigate the relationship between platelet microRNA (miR-223 and miR-96) expression and clopidogrel responsiveness in patients with coronary heart disease (CHD).

MATERIALS AND METHODS

A total of 33 consecutive non-diabetic CHD patients scheduled for percutaneous coronary intervention were enrolled. Platelet reactivity after clopidogrel loading dose (300 mg) was determined by two methods [platelet reactivity index (PRI), measured by vasodilator-stimulated phosphoprotein (VASP) phosphorylation flow cytometry and ADP-induced platelet aggregation (PAG), measured by light transmission aggregometry]. Total platelet RNA was isolated from purified platelets (CD45 magnetic bead negative selection) to quantify miR-223 and miR-96 expression by real-time PCR.

RESULTS

All subjects were dichotomized according to PRI medians (normal-responders: PRI < 56.5%, n = 17 and low-responders: PRI > 56.5%, n = 16) and PAG medians (normal-responders: PAG < 43%, n = 17 and low-responders: PAG > 43%, n = 16). Compared with PRI-determined normal-responders, miR-223 expression, but not miR-96, was significantly decreased in low-responders (P = 0.037). No differential expression of miR-223 and miR-96 was observed via PAG determination between normal- and low-responders. In addition, miR-223 expression, but not miR96, was statistically correlated with PRI (Spearman r = -0.403, P = 0.020). Stepwise binary logistic regression analysis revealed that among factors that potentially influence platelet reactivity (CYP2C19*2 loss-of-function genotypes, use of calcium channel blockers/proton-pump inhibitors, age, obesity, smoking and platelet microRNAs), decreased miR-223 expression was the only independent predictor associated with the presence of PRI-determined low responders to clopidogrel (OR 0.189, 95% CI 0.043 to 0.836, P = 0.028).

CONCLUSIONS

The present work identifies decreased platelet miR-223 expression as a novel mechanism involved in blunted platelet response to clopidogrel in a Chinese population.

摘要

目的

我们旨在研究血小板 microRNA（miR-223 和 miR-96）表达与冠心病（CHD）患者氯吡格雷反应之间的关系。

材料和方法

共纳入 33 例连续的非糖尿病 CHD 患者，计划进行经皮冠状动脉介入治疗。通过两种方法[血小板反应指数（PRI），通过血管扩张刺激磷酸蛋白（VASP）磷酸化流式细胞术测量和 ADP 诱导的血小板聚集（PAG），通过透光比浊法测量]测定氯吡格雷负荷剂量（300mg）后的血小板反应性。从纯化的血小板（CD45 磁珠阴性选择）中分离总血小板 RNA，通过实时 PCR 定量 miR-223 和 miR-96 的表达。

结果

根据 PRI 中位数（正常反应者：PRI<56.5%，n=17 和低反应者：PRI>56.5%，n=16）和 PAG 中位数（正常反应者：PAG<43%，n=17 和低反应者：PAG>43%，n=16）将所有受试者分为两组。与 PRI 确定的正常反应者相比，低反应者的 miR-223 表达（但不是 miR-96）明显降低（P=0.037）。通过 PAG 测定，在正常反应者和低反应者之间，miR-223 和 miR-96 无差异表达。此外，miR-223 表达（而不是 miR96）与 PRI 呈统计学相关（Spearman r=-0.403，P=0.020）。逐步二元逻辑回归分析显示，在可能影响血小板反应性的因素（CYP2C19*2 失活基因型、使用钙通道阻滞剂/质子泵抑制剂、年龄、肥胖、吸烟和血小板 microRNAs）中，miR-223 表达降低是与氯吡格雷反应性确定的 PRI 低反应者存在相关的唯一独立预测因子（OR 0.189，95%CI 0.043 至 0.836，P=0.028）。