血管紧张素Ⅱ诱导肾小球内皮细胞炎性损伤过程中G蛋白耦联受体激酶表达变化  

Expression of G protein-coupled receptor kinase 2 during glomerular endothelial cell inflammatory injury induced by AngiotensinⅡ

作　　者：王苗[1] 王应灯[1] 

机构地区：[1]上海交通大学医学院附属第九人民医院肾内科,上海200011

出　　处：《上海医学》2013年第3期186-189,I0003,共5页Shanghai Medical Journal

基　　金：上海市自然科学基金资助项目(092R1417400)

摘　　要：目的观察血管紧张素Ⅱ(AngⅡ)诱导肾小球内皮细胞(GENC)炎性损伤过程中内皮细胞单层通透性及G蛋白耦联受体激酶2(GRK2)表达的变化,探讨GRK2在GENC损伤中的作用及其机制。方法体外分离、培养大鼠GENC,随机分为AngⅡ组(予10mg/mL AngⅡ)、单纯滤膜组和正常对照组(予等量培养基)。体外培养12h后,应用二室弥散系统检测GENC单层通透性,反转录-聚合酶链反应(RT-PCR)检测GRK2 mRNA表达水平,Western印迹法检测GRK2蛋白表达水平。结果 AngⅡ组的单层通透率为(99.62±0.71)%,显著高于单纯滤膜组的(27.78±0.48)%和正常对照组的(34.83±1.20)%(P值均<0.01),后两组间的差异也有统计学意义(P<0.05)。AngⅡ组的GRK2 mRNA和蛋白水平均显著高于正常对照组(P值分别<0.01、0.05)。结论 10mg/mL AngⅡ诱导GENC 12h,单层通透性增加,导致细胞炎性损伤,同时GRK2 mRNA和蛋白表达水平增加,表明GRK2可能在GENC炎性损伤中发挥一定的调控作用。Objective To investigate the changes of monolayer permeability of glomerular endothelial cell （GENC） and expression of （3 protein-coupled receptor kinase 2 （GRK2） during GENC injury induced by angiotensin Ⅱ , and to explore the effect of GRK2 on GENC injury. Methods GENC was isolated and cultured from Wistar rat in vitro, and randomly divided into angiotensin group （10 mg/mL angiotensin Ⅱ ） and control group. After cultured for 12 hours in vitro, the morolayer of GENC was observed using diffusion of bovine serum albumin （Ioiotin-BSA） across the modayer. Reverse transcription PCR （RT-PCR） and Western blot were used to measure the expression of mRNA and protein of GRK2. Results The monolayer permeability in angiotensin group was significantly higher than that in membrane filter group and control group （[99.62 ± 0.71 ] %, [27.78 ± 0. 48]%, [ 34.83 ± 1. 20]%, both P〈0.01 ）. There was significant difference in permeability between membrane filter group and control group （P〈0.05）. The mRNA and protein of GRK2 in angiotensin group were significantly higher than those in control group （ P〈0.01, 0.05）. Conclusion Both the monolayer permeability of GENC and the expression of mRNA and protein of GRK2 are increased after use of 10 mg/mL angiotensin Ⅱ for 12 h, which indicates that GRK2 may regulate GENC inflammatory injury. （Shanghai Med J, 2013, 36： 186-189）

关 键 词：血管紧张素Ⅱ 肾小球内皮细胞 单层通透性 G蛋白耦联受体激酶2 

分 类 号：R363[医药卫生—病理学]