不同浓度血管紧张素Ⅱ对人脐静脉血管内皮细胞衰老变化的影响

Senescence and Influence of Human Umbilical Vein Endothelial Cells Induced by Different Concentrations Angiotensin Ⅱ

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 investigate the changes of the survival rate of Human umbilical vein endothelial cells（HUVECs）aging induced by angiotensin Ⅱ（AngⅡ） at different concentrations.Methods：HUVECs were cultured in vitro and designed randomly 5 groups as follows：control group,different concentrations groups（10-8~10-5mmol/L）.HUVECs were treated with AngII at concentrations of 0mmol/L,10-8mmol/L,10-7mmol/L,10-6mmol/L,10-5mmol/L respectively for 48h.β-gal staining was used to identify cell aging status;Flow cytometry was used for analyzing cell cycle;the cell living rate was separately observed by MTT and CCK-8.Results：After incubating the HUVECs with AngⅡ at different concentrations for 48h,the positive cell number of β-gal staining was gradually higher with the increasing concentrations（P0.01）;the cell cycle was arrested at G0/G1 phase,the relative percentage of S phase was decreased gradually;the cell living rate by AngII-induced obviously declined compared with control group;there was no significant difference in the concentration of 10-8 and 10-7mmol/L groups（P0.05）;however,10-6 and 10-5mmol/L groups has statistically significant difference（P0.05）.Conclusion：AngII induced HUVECs senescence and inhibited the cell proliferative activity according to the AngII concentrations.It means the increasing AngII concentrations could accelerate cell aging.