阿司匹林和氯吡格雷对体外血小板黏附内皮细胞基质活性的影响及其机制研究

发布时间：2018-05-20 00:36

本文选题：氯吡格雷 + 阿司匹林　；参考：《中国全科医学》2015年03期

【摘要】：目的体外研究阿司匹林和氯吡格雷对血小板黏附内皮细胞基质活性的影响及其机制,从理论上寻找两种药物联合效果优于单药的原因。方法于2013年在第三军医大学动物中心购买健康雌性SD大鼠42只,采用随机数字表法,将SD大鼠分为阿司匹林组(8只)、氯吡格雷组(8只)、阿司匹林联合氯吡格雷组(联合组,8只)、对照组(8只)以及用于原代细胞分离培养10只。分离培养原代大鼠血管内皮细胞,利用50μg/ml氧化修饰低密度脂蛋白(ox-LDL)建立受损血管内皮细胞模型并采用100 mg/kg阿司匹林、10 mg/kg氯吡格雷、100 mg/kg阿司匹林联合10 mg/kg氯吡格雷以及200μl蓖麻油制作内皮细胞基质板;血浆血小板分离制备前3 d,阿司匹林组喂养阿司匹林100mg·kg-1·d-1,氯吡格雷组喂养氯吡格雷10 mg·kg-1·d-1,联合组喂养阿司匹林100 mg·kg-1·d-1,氯吡格雷10mg·kg-1·d-1,对照组喂养蓖麻油200μl/d。采用ELISA法检测黏附内皮细胞基质上的血小板数量;采用蛋白质免疫印迹法检测阿司匹林和氯吡格雷对血管内皮细胞蛋白血栓调节蛋白(TM)、氧化型低密度脂蛋白受体1(LOX-1)、CD40表达的影响。结果经阿司匹林和氯吡格雷处理的血小板对内皮细胞黏附活性的影响:4组50、100、150、200μl血小板黏附内皮细胞基质活性比较,差异均有统计学意义(P0.05),其中氯吡格雷组和联合组50、100、150、200μl血小板黏附内皮细胞基质活性均低于对照组和阿司匹林组,阿司匹林组150μl血小板黏附内皮细胞基质活性低于对照组(P0.05)。血小板黏附经阿司匹林和氯吡格雷处理的内皮细胞基质活性的影响:4组50、100、150、200μl血小板黏附内皮细胞基质活性比较,差异均有统计学意义(P0.05),其中联合组50、100、150、200μl血小板黏附内皮细胞基质活性分别低于对照组、阿司匹林组和氯吡格雷组(P0.05)。蛋白质免疫印迹法显示,阿司匹林和氯吡格雷均能抑制由ox-LDL引起的TM表达的减少。ox-LDL能明显诱导血管内皮细胞LOX-1表达,阿司匹林能明显抑制由ox-LDL引起的LOX-1和IL-6的表达;而氯吡格雷则能明显抑制内皮细胞CD40的表达。结论阿司匹林和氯吡格雷均能从上调TM和抑制炎性因子两方面降低血小板对内皮细胞基质的黏附作用。但两者联合更能抑制由ox-LDL引起的炎性因子的表达,降低血小板对内皮细胞基质的黏附。
[Abstract]:Objective to investigate the effects of aspirin and clopidogrel on the matrix activity of platelets adhesion to endothelial cells in vitro and to find out theoretically the reason why the combination of two drugs is superior to single drug. Methods Forty-two healthy female SD rats were purchased from the Animal Center of the third military Medical University in 2013. SD rats were divided into aspirin group (n = 8), clopidogrel group (n = 8), aspirin combined with clopidogrel group (n = 8) and control group (n = 10). Primary cultured rat vascular endothelial cells were isolated and cultured. 50 渭 g/ml oxidative modification of low density lipoprotein ox-LDL (LDL) was used to establish the endothelial cell model of damaged vascular endothelial cells. The endothelial cell stroma was made with 100 mg/kg aspirin 10 mg/kg clopidogrel 100 mg/kg aspirin combined with 10 mg/kg clopidogrel and 200 渭 l castor oil. Aspirin 100mg kg-1 d-1 was given to aspirin group, clopidogrel 10 mg kg-1 d -1 to clopidogrel group, aspirin 100mg kg-1 d -1, clopidogrel 10mg kg-1 d-1 to control group, and castor oil 200 渭 l / d to control group. The effects of aspirin and clopidogrel on the expression of vascular endothelial cell protein thrombomodulin (TMN) and oxidized low density lipoprotein receptor (1LOX-1) CD40 were detected by ELISA assay and Western blot. Results the effect of aspirin and clopidogrel treatment on the adhesion activity of endothelial cells was observed. The activity of platelet adhesion endothelial cell matrix in clopidogrel group and combined group was lower than that in control group and aspirin group, and the activity of platelet adhesion endothelial cell matrix in aspirin group was lower than that in control group (P 0.05). Effect of Aspirin and Clopidogrel treatment on Platelet Adhesion to Endothelial Cell Matrix activity comparison of 50100150200 渭 l Platelet Adhesion Endothelial Cell Matrix activity, The activity of platelet adhesion to endothelial cell matrix in combined group was lower than that in control group, and that in aspirin group and clopidogrel group was lower than that in aspirin group and clopidogrel group. Western blot analysis showed that both aspirin and clopidogrel could inhibit the decrease of TM expression induced by ox-LDL. Ox-LDL could significantly induce the expression of LOX-1 in vascular endothelial cells, and aspirin could inhibit the expression of LOX-1 and IL-6 induced by ox-LDL. Clopidogrel inhibited the expression of CD40 in endothelial cells. Conclusion Aspirin and clopidogrel can both up-regulate TM and inhibit inflammatory factors to reduce platelet adhesion to endothelial cell matrix. However, the combination of the two can inhibit the expression of inflammatory factors induced by ox-LDL and reduce the adhesion of platelets to endothelial cell matrix.
【作者单位】：贵州省贵阳市第一人民医院;第三军医大学附属新桥医院;
【基金】：贵州省科技厅社会发展项目(黔科合SY[2010]3087)
【分类号】：R965

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