摘要
目的 探究腺苷二磷酸（ADP）监测阿司匹林抗血小板聚集的作用及相关的信号转导通路,为抗血小板聚集药物的筛选提供指导。方法 SD大鼠灌胃给药（0.5%CMC-Na,5 ml/kg;阿司匹林20、50和100 mg/kg;普拉格雷25 mg/kg）,均为一日1次,共4 d。最后一次给药后1 h,大鼠经水合氯醛麻醉,腹主动脉采血,低速离心,收集上层液（富血小板血浆）。应用不同浓度（0、0.1、0.3、1和10μmol/L）的ADP对阿司匹林抗血小板聚集的作用进行监测。利用富血小板血浆进行蛋白质印迹试验,观察相关蛋白激酶的激活情况。结果 不同剂量阿司匹林对ADP诱导血小板聚集的抑制率随着ADP浓度的增加（0-10μmol/L）呈先升高后降低的趋势,这种现象与阿司匹林剂量无关。在ADP浓度为0.3μmol/L时,阿司匹林低（20 mg/kg）、中（50 mg/kg）及高剂量（100 mg/kg）组的血小板聚集抑制率均为同组中最高（与阴性对照组比较,P〈0.01）。而在ADP浓度为10μmol/L时,阿司匹林的血小板聚集抑制率均小于10%,对应普拉格雷组（25 mg/kg）的血小板聚集抑制率为（76.72±7.83）%;蛋白质印迹试验结果表明,不同剂量阿司匹林均可引起ERK2显著磷酸化,且ERK2磷酸化程度随阿司匹林剂量的增加有减弱趋势。结论 低剂量ADP（0.3μmol/L）用于监测阿司匹林的抗血小板聚集作用更为可靠。
Objective To investigate the aspirin-mediated platelet aggregation inhibition through adenosine diphosphate （ADP） detection, and study the possible underlying mechanisms. Methods SD rats were pretreated with 0.5% CMC-Na, aspirin （20, 50 and 100 mg/kg） and prasugrel （25 mg/kg） by oral gavage once daily for 4 days. In one hour after the last dosing on the 4th day, the rats were anesthetized with chloral hydrate and the whole blood were collected from the abdominal aorta and centrifuged. The supernatant was used for platelet aggregation analysis. Different doses of ADP （final concentration were 0, 0.1, 0.3, 1 and 10 μmol/L） were used to monitor the platelet reactivity after the aspirin treatment. Platelets were also homogenized and prepared for Western blotting in order to detect related protein kinase activation. Results There was a bell-shape curve for the dose-response correlation of the inhibition percentage of ADP-induced platelet aggregation in all the three aspirin-treated groups, which showed an increase of platelet aggregation inhibition in the beginning and then a decrease later. This response seemed to be unrelated to the dosage of aspirin. Aspirin at the three doses （20, 50 and 100 mg/kg） induced the maximal inhibition response of the platelet aggregation by ADP at the dose of 0.3 gmol/L. But ADP at 10 gmol/L showed a weak inhibition of ADP-induced platelet aggregation （〈10%）. In Western blotting test, a significant increase of ERK2 phosphorylation following aspirin treatment with all the three doses were observed. Moreover, the ERK2 phosphorylation had decreased tendency as increasing aspirin dose. Conclusion The lower dose of ADP （0.3 μmol/L） can monitor more reliably the aspirin-mediated inhibition of platelet aggregation.
出处
《世界临床药物》
CAS
2016年第2期124-128,共5页
World Clinical Drug
基金
上海市浦江人才计划(编号:13PJ1433100)
留学人员科技活动项目资助
上海市生物物质成药性评价专业技术服务平台(编号:15DZ2291700)
