血管紧张素(1-7)通过p53/Drp1轴对血管紧张素Ⅱ诱导的血管内皮细胞衰老的影响

Effect of angiotensin(1-7)on angiotensinⅡ-induced vascular endothelial cell senescence by the p53/Drp1 pathway

目的观察血管紧张素(1-7)[Ang(1-7)]对血管紧张素Ⅱ(AngⅡ)诱导的人脐静脉内皮细胞(HUVEC)衰老的作用及机制。方法体外用含有10%胎牛血清(FBS)的DMEM培养基培养HUVEC 48 h,随机分为对照组、Ang(1-7)组(1μmol/L)、AngⅡ组(1μmol/L)和AngⅡ+Ang(1-7)组。通过细胞衰老β-半乳糖苷酶(SA-β-Gal)染色试剂盒检测各组衰老细胞数量(光学显微镜观察),通过活性氧检测试剂盒测定各组细胞活性氧(ROS)的水平,通过Western blot检测各组细胞p53和动力蛋白相关蛋白1(Drp1)的表达。结果与对照组比较,AngⅡ组SA-β-Gal染色阳性细胞明显增多(P<0.001),细胞ROS水平增加(P<0.001),p53及Drp1蛋白表达量明显增加(P<0.01)。与AngⅡ组比较,AngⅡ+Ang(1-7)组SA-β-Gal染色阳性细胞率(P<0.01)、细胞ROS水平(P<0.001)、p53及Drp1蛋白表达量(P<0.05)均降低。结论Ang(1-7)可能通过影响p53/Drp1通路,抑制HUVEC内ROS生成,减轻AngⅡ诱导的HUVEC衰老。

Aim The purpose of this study was to observe the effect and mechanism of angiotensin(1-7)(Ang(1-7))on angiotensinⅡ(AngⅡ)-induced senescence of human umbilical vein endothelial cell(HUVEC).Methods HUVEC were cultured in vitro with DMEM high glucose medium containing 10%fetal bovine serum(FBS)for 48 hours and randomly divided into control group,Ang(1-7)group(1μmol/L),AngⅡgroup(1μmol/L),and AngⅡ+Ang(1-7)group.Cell senescenceβ-galactosidase(SA-β-Gal)staining kits were used to detect the number of senescent cell in each group(observed under an optical microscope);reactive oxygen species(ROS)levels were measured using reactive oxygen detection kits,and p53 and dynamin related protein-1(Drp1)expression in each group were detected by Western blot.Results Compared with the control group,both SA-β-Galpositive cell and ROS levels in the AngⅡgroup were significantly increased(P<0.001),and p53 and Drp1 protein expression in the cell were significantly increased(P<0.01).Compared with AngⅡgroup,SA-β-Galpositive cell(P<0.01),ROS levels(P<0.001),p53 and Drp1 protein expression(P<0.05)were decreased in AngⅡ+Ang(1-7)group.Conclusion Ang(1-7)may inhibit the ROS generation in HUVEC by affecting the p53/Drp1 pathway,and reverse the aging of HUVEC induced by AngⅡ.