G蛋白偶联受体激酶4突变A142V基因对大鼠血管平滑肌细胞AT_1受体的影响  

Regulation of human G-coupled protein kinase 4 A142V on AT_1 receptor in rat vascular smooth muscle cells

作　　者：邓昆[1,2] 刘莉[1] 陈彩宇[1] 陈垦[1] 王微[1] 周永巧[1] 何多芬[1] 曾春雨[1] 

机构地区：[1]第三军医大学大坪医院野战外科研究所心血管内科/重庆心血管病研究所 [2]77263部队医院

出　　处：《重庆医学》2013年第33期3977-3979,共3页Chongqing medicine

基　　金：国家杰出青年科学基金资助项目(30925018);国家自然科学基金资助项目(31130029;81070559;81100500)

摘　　要：目的研究人G蛋白偶联受体激酶4(GRK4)变异体A142V对大鼠血管平滑肌细胞(VMSCs)血管紧张素Ⅱ1型(AT1)受体及其介导的VMSCs增殖的影响,以期了解GRK4引起原发性高血压的原因。方法构建与增强型绿色荧光蛋白(EGFP)融合表达的慢病毒载体,包装慢病毒,感染A10细胞并进行鉴定;免疫印迹方法检测AT1受体蛋白变化;采用分光光度法检测GRK4活性,免疫共沉淀检测GRK4和AT1受体的共连接;[3 H]胸腺嘧啶掺入的方法检测增殖变化。结果转染hGRK4γA142V细胞的GRK4酶活性显著升高,AT1受体表达显著升高,GRK4和AT1受体共连接作用降低,AngⅡ刺激细胞增殖效应显著增强。结论转染hGRK4变异体A142V增加GRK4活性,引起AT1受体的功能增强和VMSCs的增殖作用增加。Objective To study the effect of human G-coupled protein kinase 4（GRK4） A142V overexpression on angiotensin Ⅱ1 type（AT1 ） receptor and its-mediated proliferation of rat vascular smooth muscle cells .Methods We constructed a lentiviral vec-tor carrying human GRK4-EGFP gene and observed its expression in A10 cells .Expression of AT1 receptor were determined by im-munoblotting ,GRK4 activity were checked by spectrophotometry ;the linkage between GRK4 and AT1 receptor were determined by co-immunoprecipitation .[3 H] thymidine incorporation was used to detect changes of cell proliferation .Results As compared with the control cells ,A142V-transfected cells had higher GRK4 activity and higher AT1 receptor expression ;there was linkage between GRK4 and AT1 receptor ,the co-immunoprecipitation levels were lower in A142V cells .The basal levels of VSMC proliferation was higher in A142V cells ,Ang Ⅱ increased VSMC proliferation to a greater extent in A 142V cells .Conclusion GRK4 A142V ,via in-creasing GRK4 activity ,increases AT1 receptor expression and function in vascular smooth muscle cell proliferation .

关 键 词：高血压 G蛋白偶联受体激酶4 受体 血管紧张素 1型 血管平滑肌细胞 

分 类 号：R544.1[医药卫生—心血管疾病]