益气活血中药配伍双联抗血小板药物对人脐静脉内皮细胞损伤和内皮血小板黏附的影响  

Chinese Herbal Compounds for Supplementing Qi and Activating Blood Circulation Combined with Dual Antiplatelet Drugs Alleviated Human Umbilical Vein Endothelial Cell Injury and Platelet Adhesion via Up-regulation of PI3K/Akt Pathway

作　　者：王铭铭[1] 薛梅[1] 杨琳[1] 缪宇[1] 许勇钢[1] 寇娜[2] 曲华[2] 史大卓[1] 

机构地区：[1]中国中医科学院西苑医院心血管科,北京100091 [2]北京中医药大学研究生院,北京100029

出　　处：《中国中西医结合杂志》2016年第7期842-848,共7页Chinese Journal of Integrated Traditional and Western Medicine

基　　金：国家自然科学基金青年基金项目(No.81102722);国家自然科学基金面上项目(No.81273933);北京市自然科学基金青年基金资助项目(No.7144240)

摘　　要：目的基于PI3K/Akt通路研究益气活血中药联合双联抗血小板药物对氧化低密度脂蛋白(oxidized low density lipoprotein,ox-LDL)诱导的人脐静脉内皮细胞(human umbilical vein endothelial cell,HUVEC)损伤及损伤内皮诱导的血小板黏附的作用及机制。方法将HUVEC随机分为空白组、模型(80 mg/L ox-LDL)组、双抗(15μg/m L阿司匹林+10μg/m L氯吡格雷+80 mg/L ox-LDL)组、西洋参茎叶总皂苷(Panax Quinquefolium saponins,PQS,160μg/m L)+三七总皂苷(Panax Notoginseng saponins,PNS,160μg/m L)+双抗组和LY294002(30μg/m L)+PQS+PNS+双抗组,共5组。利用流式细胞术检测HUVEC凋亡率和内皮-血小板黏附情况;生化法检测HUVEC上清液乳酸脱氢酶浓度(lactate dehydrogenase,LDH);放射免疫法检测HUVEC上清液细胞间黏附分子浓度(intercellular adhesion molecule,ICAM);Western blot检测HUVEC中p-PI3K、p-Akt蛋白表达。结果与空白组比较,模型组HUVEC凋亡率、平均荧光强度(mean fluorescence indicator,MFI)、LDH、ICAM浓度均升高(P<0.05),HUVEC p-Akt蛋白表达降低(P<0.05)。与模型组比较,双抗组HUVEC凋亡率和LDH浓度均升高(P<0.05),MFI和ICAM浓度均降低(P<0.05);PQS+PNS+双抗组凋亡率、MFI、LDH、ICAM均降低(P<0.05),PQS+PNS+双抗组HUVEC中p-Akt表达增加(P<0.05)。与双抗组比较,PQS+PNS+双抗组HUVEC凋亡率、MFI、LDH及ICAM均明显降低(P<0.05),HUVEC中p-Akt表达升高(P<0.05);加入PI3K特异性抑制剂LY294002后,HUVEC中p-Akt表达被抑制,益气活血中药配伍双抗发挥的上述保护作用均消失。结论益气活血中药配伍双抗通过上调内皮细胞PI3K/Akt通路,减轻ox-LDL诱导的内皮细胞凋亡以及损伤内皮诱导的血小板黏附。Objective To observe the effect and underlying mechanism of Chinese herbal compound（CHC）for supplementing qi and activating blood circulation（SQABC） combined with dual antiplatelet drugs（DA） on oxidized low density lipoprotein（ox-LDL） induced human umbilical vein endothelial cell（HUVEC） injury and platelet adhesion evoked by injured endothelial cells（ECs） based on PI3K/Akt signaling pathway. Methods HUVECs were randomly divided into 5 groups, i.e., the blank control group, the model group（80 mg/L ox-LDL）, the DA group（15 μg/m L aspirin ＋ 10 μg/ m L clopidogrel ＋ 80 mg/L ox-LDL）, the Panax Quinquefolium saponins（PQS, 160 μg/m L） ＋ Panax Notoginseng saponins（PNS, 160 μg/m L） ＋ DA group, the LY294002（30 μg/m L） ＋ PQS ＋ PNS ＋ DA group. HUVEC ap-optosis rate and platelet adhesion to HUVECs were detected by flow cytometry. Concentration of lactate dehydrogenase（LDH） in HUVEC supernatant was detected by biochemical assay. Concentration of intercellular adhesion molecular（ICAM） was detected by radioimmunoassay. Protein expressions of p-PI3 K and p-Akt in HUVECs were detected by Western blot. Results Compared with the blank control group, the apoptosis rate of HUVECs, mean fluorescence indicator（MFI）, concentrations of both LDH and ICAM increased（P 〈0. 05）, and p-Akt protein expression decreased（P 〈0. 05） in the model group. Compared with the model group, the apoptosis rate of HUVECs and LDH concentration increased（P 〈0. 05）, concentrations of MFI and ICAM obviously decreased（P 〈0. 05） in the DA group. The apoptosis rate, MFI, concentrations of both LDH and ICAM all decreased in the PQS ＋ PNS ＋ DA group（P 〈0. 05）. pAkt protein expression in HUVECs obviously increased in the PQS ＋ PNS ＋ DA group（P 〈0. 05）. Compared with the DA group, HUVEC apoptosis rate, MFI, concentrations of both LDH and ICAM in supernatant obviously decreased, pAkt expression in HUVECs increased in the PQS ＋ PNS ＋ DA group（all P

关 键 词：益气活血 内皮细胞 血小板黏附 双联抗血小板药物 PI3K 

分 类 号：R285[医药卫生—中药学]