Dietary Nitrate Supplementation Reduces Circulating Platelet-Derived Extracellular Vesicles in Coronary Artery Disease Patients on Clopidogrel Therapy: A Randomised, Double-Blind, Placebo-Controlled Study
Dietary Nitrate Supplementation Reduces Circulating Platelet-Derived Extracellular Vesicles in Coronary Artery Disease Patients on Clopidogrel Therapy: A Randomised, Double-Blind, Placebo-Controlled Study
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DOI：
10.1160/th17-06-0394
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发表时间：
2018-01-01
影响因子：
6.7
通讯作者：
James, Philip E.
中科院分区：
文献类型：
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作者：
Burnley-Hall, Nicholas;Abdul, Fairoz;James, Philip E.
细胞外小泡(EVS)参与心血管疾病(CVD)的发病机制。具体地说，血小板衍生的EV是高度促凝血的，促进凝血酶的生成和纤维蛋白凝块的形成。硝酸盐补充剂通过增加一氧化氮(NO)的生物利用度在心血管疾病中发挥有益的作用。氯吡格雷能够在亚硝酸盐和低pH条件下产生NO供体化合物，如S亚硝硫醇(RSNO)。本研究的目的是评估补充硝酸盐并与不加氯吡格雷治疗对冠心病(CAD)患者循环EVS的影响。在这项随机、双盲、安慰剂对照研究中，接受氯吡格雷治疗(n=10)或不接受氯吡格雷治疗(n=10)的冠心病患者接受了饮食硝酸盐补充剂(SIS硝酸盐凝胶)或相同的安慰剂。用基于臭氧的化学发光法和多电极聚集法测定NO代谢产物和血小板活化。利用纳米粒子跟踪分析和时间分辨荧光分析确定EV的浓度和来源。补充硝酸盐后，与安慰剂组相比，氯吡格雷组血浆RSNO升高(4.7+/-0.8vs0.2+/-0.5 nM)，凝血酶受体介导的血小板聚集率降低(-19.9+/-6.0vs4.0+/-6.4U)。与安慰剂组相比，本组患者的循环EVS显著降低(-1.183E(11)+/-3.15E(10)vs-9.93E(9)+/-1.84E(10)Evs/mL)，尤其是CD41+EVS的比例(-2,120+/-728vs 235+/-436RFU[相对荧光单位])。体外实验表明，氯吡格雷-SNO可直接降低血小板EV(6.209E(10)+/-4.074E(9)vs 3.94E(11)+/-1.91E(10)EVS/mL)。总之，服用氯吡格雷的冠心病患者补充硝酸盐减少了血小板衍生的EVS，增加了患者对氯吡格雷的反应性。补充硝酸盐可能代表了一种降低冠心病患者血栓形成风险的新方法。
Extracellular vesicles (EVs) are implicated in the pathogenesis of cardiovascular disease (CVD). Specifically, platelet-derived EVs are highly pro-coagulant, promoting thrombin generation and fibrin clot formation. Nitrate supplementation exerts beneficial effects in CVD, via an increase in nitric oxide (NO) bioavailability. Clopidogrel is capable of producing NO-donating compounds, such as S-nitrosothiols (RSNO) in the presence of nitrite and low pH. The aim this study was to assess the effect of nitrate supplementation with versus without clopidogrel therapy on circulating EVs in coronary artery disease (CAD) patients. In this randomized, double-blind, placebo-controlled study, CAD patients with (n = 10) or without (n = 10) clopidogrel therapy received a dietary nitrate supplement (SiS nitrate gel) or identical placebo. NO metabolites and platelet activation were measured using ozone-based chemiluminescence and multiple electrode aggregometry. EV concentration and origin were determined using nanoparticle tracking analysis and time-resolved fluorescence. Following nitrate supplementation, plasma RSNO was elevated (4.7 +/- 0.8 vs 0.2 +/- 0.5 nM) and thrombin-receptor mediated platelet aggregation was reduced (-19.9 +/- 6.0 vs 4.0 +/- 6.4 U) only in the clopidogrel group compared with placebo. Circulating EVs were significantly reduced in this group (-1.183e(11) +/- 3.15e(10) vs -9.93e(9) +/- 1.84e(10) EVs/mL), specifically the proportion of CD41+ EVs (-2,120 +/- 728 vs 235 +/- 436 RFU [relative fluorescence unit]) compared with placebo. In vitro experiments demonstrated clopidogrel-SNO can reduce platelet-EV directly (6.209e(10) +/- 4.074e(9) vs 3.94e(11) +/- 1.91e(10) EVs/mL). In conclusion, nitrate supplementation reduces platelet-derived EVs in CAD patients on clopidogrel therapy, increasing patient responsiveness to clopidogrel. Nitrate supplementation may represent a novel approach to moderating the risk of thrombus formation in CAD patients.