血管紧张素Ⅱ诱导血管平滑肌细胞肥厚过程中MAPK对细胞周期素依赖性激酶抑制因子的调节作用  

Regulation of cyclin-dependent kinase inhibitors by mitogen-activated protein kinase in angiotensinⅡ-stimulated vascular smooth muscle cell hypertrophy

作　　者：刘煜[1] 许顶立[1] 汪明慧[1] 欧阳平[1] 赖文岩[1] 

机构地区：[1]第一军医大学南方医院心血管内科,广东广州510515

出　　处：《第一军医大学学报》2002年第2期114-116,共3页Journal of First Military Medical University

基　　金：国家自然科学基金(39770325);国家重点基础研究发展规划(G2000056905)

摘　　要：目的观察在血管紧张素Ⅱ(AngⅡ)诱导血管平滑肌细胞(VSMC)肥厚过程中,丝裂原活化蛋白激酶(MAPK)活性和细胞周期素依赖性激酶抑制因子p27、p21、p57蛋白表达量的变化,探讨MAPK对细胞周期素依赖性激酶抑制因子的调节作用。方法分离SD大鼠主动脉中层平滑肌,贴壁法培养平滑肌细胞,无血清培养基培养静止后,加入AngⅡ1×10-6 mol/L和/或豆蔻酸佛波酰乙酯(PMA)20nmol/L,以无血清培养基培养的VSMC作对照。在刺激后90min收集细胞,用免疫沉淀法检测MAPK活性的改变。在刺激后6和24h收集细胞,用Western blotting法检测p27、p57和p21蛋白表达量。结果AngⅡ可以使MAPK活性升高,但其升高幅度较低(仅较对照组增加了138%),未能抑制p27蛋白的表达,不能使VSMC增殖。PMA和AngⅡ共同作用时,可以轻度抑制p27蛋白的表达,但仍未能使VSMC增殖。结论MAPK通路是VSMC胞外增殖肥厚刺激信号进入核内的一条重要信息传导通路。Objective To investigatetheroleof mitogen-activatedproteinkinase(MAPK)in theregulationof cyclin-depen-dentkinaseinhibitors(CDKI)in theprocessof vascularsmoothmusclecell(VSMC)hypertrophyinducedby angiotensinⅡstimulation.Methods Themediallayerof maleSD rataortawasisolatedforVSMCculture.Afterculturedinserum-free mediumto arrestthecellgrowth,VSMCswerestimulatedwithangiotensinⅡ(1×10 -6 mol/L)or/andphorbolmyristateac-etate(PMA,20nmol/L),withthecellsculturedin serum-freemediumservingas control.MAPKactivityof thecellswas assayed90minafterstimulationwithimmunoprecipitationtest,andtheexpressionlevelsof CDKIp27,p57andp21were determinedby Westernblotting6and24h afterstimulationrepectively.Results Comparedwiththecontrollevel,MAPK activityof theVSMCswasup-regulatedby238%by treatmentwithangiotensinⅡalonewhich,however,didnotinhibit p27expressionor induceVSMCproliferation.CostimulationwithangiotensinⅡandPMAslightlyinhibitedp27expres-sionbutVSMCproliferationwas stillnotobserved.Conclusion MAPKpathwayis an importantchannelfor extracellular proliferativeandhypertrophicsignaltransductionintothenucleusof VSMCs.

关 键 词：血管紧张素Ⅱ 丝裂原活化蛋白激酶 细胞周期蛋白质依赖性激酶类 肥大性心肌病 血管平滑肌 

分 类 号：R540.2[医药卫生—心血管疾病]