血管紧张素Ⅱ对血管内皮细胞凋亡及p38丝裂素活化蛋白激酶表达的影响  

Effect of AngiotensinⅡ on Apoptosis and Expression of p38 Mitogen-Activated Protein Kinase in Vascular Endothelial Cells

作　　者：单海燕[1] 刘姝[1] 胡翠竹[1] 李效丽[1] 白小涓[1] 陈香美[2] 

机构地区：[1]中国医科大学附属第一医院老年心血管科,辽宁省沈阳市110001 [2]中国人民解放军总院肾病中心,北京市100853

出　　处：《中国动脉硬化杂志》2011年第1期13-17,共5页Chinese Journal of Arteriosclerosis

基　　金：中华医学会临床医学科研专项资金-动脉粥样硬化研究资金资助(09010530208);辽宁省科学技术研究项目(20091104);沈阳市科学技术计划项目(F10-205-1-44)

摘　　要：目的探讨血管紧张素Ⅱ对人脐静脉内皮细胞凋亡的影响以及p38丝裂素活化蛋白激酶表达的变化。方法制备血管紧张素ⅡRPM I1640培养液培养人脐静脉内皮细胞,通过吖啶橙荧光染色观察细胞形态学的变化、流式细胞仪检测细胞凋亡率,利用逆转录聚合酶链反应和免疫印迹法分析凋亡调控基因Bc l-2 mRNA及蛋白表达水平,通过特异性磷酸化p38丝裂素活化蛋白激酶抗体采用免疫印迹法检测p38丝裂素活化蛋白激酶活性。结果血管紧张素Ⅱ能诱导内皮细胞凋亡,荧光显微镜可见明显的细胞凋亡(32.76%±2.98%比2.14%±0.36%,P<0.01);流式细胞仪检测AnnexinV-FITC/PI双染色的血管紧张素Ⅱ诱导人脐静脉内皮细胞中,早中期凋亡细胞明显增加,其细胞凋亡率为37.4%±1.6%(对照组为10.2%±1.8%)。与对照组相比,6 h、12 h、18 h及24 h Bc l-2 mR-NA及蛋白表达水平明显降低(P<0.05),p38丝裂素活化蛋白激酶磷酸化水平于18 h达到最高峰(P<0.01)。结论血管紧张素Ⅱ诱导内皮细胞的凋亡可能与p38丝裂素活化蛋白激酶对Bc l-2 mRNA及蛋白表达影响有关。Aim To investigate the effects of angiotensinⅡ（AngⅡ） on apoptosis,and phosphorylation of p38 mitogen-activated protein kinase in endothelial cell,and its possible action mechanism. Method Human umbilical vein endothelial cells（HUVEC） were cultured in vitro and intervened by AngⅡ.HUVEC were divided into control group and AngⅡ group（stimulated by AngⅡ10-6 mol/L for 24 h）,morphologic changes and percentage of apoptosis were assayed with acridine orange fluorescence staining.The early stage apoptosis was detected by flow cytometery with Annexin V-FITC/PI double staining.The expression of apoptosis-association gene Bcl-2 was detected by RT-PCR and Western blotting at different time points.By means of Western blotting,the activation of p38MAPK was observed at different time points. Results 10-6 mol/L AngⅡ stimulated cell apoptosis.The percentage of apoptotic cells in AngⅡ-stimulated cells was significantly increased compared to that in the control cells（32.76%±2.98% vs 2.14%±0.36%,P0.01） by using acridine orange fluorescence staining.The early-metaphase apoptotic rate was significantly increased in AngⅡ-stimulated cells compared to the control cells（37.4%±1.6% vs 10.2%±1.8%,P0.01） by Annexin V-FITC/PI double staining flow cytometry.Bcl-2 mRNA and protein expression decreased markedly（P0.05）,the activation of p38MAPK began to increase and reach the peak at 18 h（P0.01）. Conclusions Cell apoptosis is possibly an impor tant factor for atherosclerosis.One of its molecular mechanisms might be associated with decreasing the expression level of Bcl-2.There is a probability that activated p38MAPK signal pathway is involved in the process of pathologic and physiologic reaction in the apoptosis of endothelial cell induced by AngⅡ.

关 键 词：血管紧张素Ⅱ 内皮细胞 细胞凋亡 P38丝裂素活化蛋白激酶 

分 类 号：R363[医药卫生—病理学]