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目的观察整合素αvβ3在血管紧张素Ⅱ(AngⅡ)诱导的人脐静脉内皮细胞(HUVECs)衰老中的变化。方法体外培养HUVECs,采用CCK-8法检测细胞存活率,用AngⅡ(终浓度10-6mol/L)干预,分为实验对照组、AngⅡ诱导组。以衰老相关β-半乳糖苷酶活性和细胞增殖能力两种衰老标志物为主要观察指标。其中衰老相关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周期来反应细胞的增殖能力;利用Western印迹法分析AngⅡ诱导HUVECs 0、12、24、36、48 h的整合素αvβ3表达的时间效应关系。结果与对照组相比,10-6 mol/L AngⅡ诱导组存活的细胞数为对照组的(77.15±6.83)%;(81.80±0.92)%的细胞呈现β-半乳糖苷酶阳性染色,流式细胞仪检测细胞周期停滞于G0～G1,证实细胞衰老;AngⅡ呈时间依赖性上调整合素αvβ3表达。结论 AngⅡ可以诱导HUVECs衰老,其机制可能与上调衰老细胞整合素αvβ3表达有关。
Objective To observe the changes of integrin αvβ3 in senescence induced by angiotensin Ⅱ (AngⅡ) in human umbilical vein endothelial cells (HUVECs). Methods HUVECs were cultured in vitro. Cell viability was detected by CCK-8 assay. AngⅡ (final concentration 10-6 mol / L) was used to induce cell viability. The cells were divided into experimental group and AngⅡ-induced group. The aging-related β-galactosidase activity and cell proliferation of two kinds of aging markers as the main observation. The senescence-relatedβ-galactosidase activity was detected by immunochemical staining, and the cell cycle was detected by flow cytometry to detect the cell proliferation. Western blotting was used to analyze the effect of AngⅡ on the integration of HUVECs at 0, 12, 24, 36 and 48 h Time-dependent effect of αvβ3 expression. Results Compared with the control group, the number of surviving cells in the 10-6 mol / L AngⅡinduced group was (77.15 ± 6.83)% of the control group; (81.80 ± 0.92)% of the cells showed β-galactosidase positive staining, Cytometry showed that cell cycle arrest at G0 ~ G1, which confirmed cellular senescence; AngⅡ up-regulated the expression of integrin αvβ3 in a time-dependent manner. Conclusion Ang Ⅱ can induce the senescence of HUVECs, and its mechanism may be related to up-regulating the expression of integrin αvβ3 in senescent cells.