Senescence of endothelial cells and gene expression associated with apoptosis induced by angiotensinⅡ / 中国病理生理杂志

ABSTRACT

AIM:To study the senescence of human umbilical vein endothelial cells(HUVECs) and Bcl-2,Bax gene expression associated with apoptosis induced by angiotensinⅡ(AngⅡ).METHODS:HUVECs were cultured in vitro and the cell viability was observed by methyl thiazolyl tetrazolium(MTT).HUVECs were intervened by AngⅡ and valsartan(AngⅡ type 1 receptor blocking) and divided into 3 groups:the control group,AngⅡ group(stimulated with AngⅡ10-6mol/L for 48 h),valsartan group(valsartan was added to cells 1 h before 10-6mol/L AngⅡ treatment).?-gal staining aod cell cycle analysis were used to identify the cell aging status.Morphologic changes and percentage of apoptosis were assayed with Hoechst33258 under fluorescent microscope.The expressions of Bcl-2 and Bax,and the apoptosis-associated genes were detected by immunocytochemical staining,RT-PCR and Western blotting.RESULTS:The cell viability by AngⅡ-induced cells was(81.9%?4.1)%,the positive cell number of ?-gal staining was significantly higher in AngⅡ-induced cells(80.10%?6.81)% than that in the control cells.The cell cycle was at G0-G1(91.36%?6.45)%,the apoptotic cells significantly increased(31.84?2.86)% under fluorescent microscope.In valsartan group,Bcl-2 mRNA and protein expression increased markedly(P