目的 观察血管紧张素Ⅱ( AngⅡ)诱导的人脐静脉内皮细胞(HUVEC)衰老及对细胞骨架的影响.方法 体外培养HUVEC,采用CCK-8法检测细胞存活率,用AngⅡ干预,分为对照组、10-6 mol/L AngⅡ诱导组(AngⅡ组).主要观察衰老相关β-半乳糖苷酶活性和细胞的增殖能力.β-半乳糖苷酶活性采用免疫化学染色方法,流式细胞术检测细胞增殖能力;考马斯亮蓝染色显示内皮细胞骨架,利用Western blot印迹法分析波形蛋白表达的变化.结果 与10-6 mol/L AngⅡ细胞存活率(77.15±6.83)％比较,10 5 mol/L AngⅡ存活率(55.61±7.92)％明显降低(P＜0.01);AngⅡ组β-半乳糖苷酶阳性染色细胞数目明显增加(P＜0.001),流式细胞仪检测细胞周期停滞于G0/G1期[(80.11±7.92)％];AngⅡ组细胞骨架及波形蛋白较对照组明显降解.结论 AngⅡ可以诱导HUVEC衰老,其分子机制可能与细胞骨架蛋白裂解有关.%Objective To observe the angiotensin H (Angll )-induced aging of human umbilical vein endothelial cells(HUVEC) and its effect on cytoskeleton. Methods Survival rate of in vitro cultured HUVEC was detected by CCK-8 assay. HUVEC were divided into control group and Ang H (at the concentration of 10~6 mol/L) treatment group. Ageing-related (3-galactosidase activity was assayed with immunocytochemical staining and proliferation of HUVEC was detected by flow cy-tometry. Endothelial skeleton was displayed with Coomassie bright blue staining. Expression of vimentin was detected by Western blot. Results The survival rate of HUVEC was 77. 15% + 6. 83 % in Angll treatment group,which was higher than that in control group. The positive number of (3-galactosidase-stained HUVEC increased significantly in Ang fj treatment group (P < 0. 01). Flow cytometry showed that the cell cycle of HUVEC was at G0/GK80. 11%?. 92%). The expression level of cytoskeleton and vimentin protein was significantly lower in Ang TJ treatment group than in control group. Conclusion Angll induces aging of HUVEC and its molecular mechanism may be related with the degradation of cytoskeletal protein.
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