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目的:观察不同浓度血管紧张素Ⅱ(AngⅡ)诱导人脐静脉血管内皮细胞(HUVECs)衰老及细胞存活率的变化情况。方法:体外培养HUVEC,随机分为对照组和含不同浓度AngⅡ组(10-8～10-5mmol/L)共5组,用含不同浓度AngⅡ的培养基孵育细胞48h,β-半乳糖苷酶染色法鉴定内皮细胞衰老状态;流式细胞技术分析细胞周期变化;分别用MTT法和CCK-8法检测细胞存活率变化情况。结果:AngⅡ培养细胞48h后,β-gal阳性染色率随着AngⅡ浓度的增加逐渐增多(P均<0.01);细胞周期多停滞于G0/G1期,S期细胞逐渐减少;细胞存活率明显下降;与对照组相比,10-8和10-7 mmol/L AngII组无明显差异(P>0.05),10-6和10-5 mmol/LAngⅡ组有统计学差异(P<0.05)。结论:AngII诱导HUVEC衰老并呈剂量依赖性的抑制细胞增殖,提示细胞衰老程度随AngII浓度增加而加重。
Objective: To observe the changes of senescence and cell viability induced by different concentrations of angiotensin Ⅱ in human umbilical vein endothelial cells (HUVECs). Methods: HUVECs were cultured in vitro and randomly divided into control group and AngⅡ group (10-8-10-5mmol / L) for 5 days. Cells were incubated with different concentrations of AngⅡ for 48h, and β-galactosidase The aging of endothelial cells was identified by staining, the cell cycle was analyzed by flow cytometry, and the cell viability was detected by MTT assay and CCK-8 assay. Results: After 48 hours of AngⅡtreatment, the positive staining rate of β-gal gradually increased with the increase of AngⅡ concentration (all P <0.01). The cell cycle arrested in G0 / G1 phase and decreased gradually in S phase. The cell survival rate decreased significantly Compared with the control group, there was no significant difference between the 10-8 and 10-7 mmol / L AngII groups (P> 0.05), but there was a significant difference between the 10-6 and 10-5 mmol / LAngⅡ groups (P <0.05). CONCLUSION: AngII induced the senescence of HUVEC and inhibited cell proliferation in a dose-dependent manner, suggesting that the degree of cell senescence aggravated with the increase of AngII concentration.