CAJ | 학술논문

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 A T1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。
목적：연구인G단백우련수체격매4（GRK4）변이체 A142V대대서혈관평활기세포（VMSCs）혈관긴장소Ⅱ1형（AT1）수체급기개도적VMSCs증식적영향，이기료해 GRK4인기원발성고혈압적원인。방법구건여증강형록색형광단백（EGFP）융합표체적만병독재체，포장만병독，감염A10세포병진행감정；면역인적방법검측 AT1수체단백변화；채용분광광도법검측GRK4활성，면역공침정검측 GRK4화 A T1수체적공련접；[3 H ]흉선밀정참입적방법검측증식변화。결과전염hGRK4γA142V세포적GRK4매활성현저승고，AT1수체표체현저승고，GRK4화AT1수체공련접작용강저，AngⅡ자격세포증식효응현저증강。결론전염hGRK4변이체A142V증가GRK4활성，인기AT1수체적공능증강화VMSCs적증식작용증가。
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 .