血管紧张素(1-7)对ox-LDL诱导人脐静脉内皮细胞凋亡和炎症反应的影响

Effect of Angiotension(1-7)on the apoptosis and inflammation of human umbilical vein endothelial cells induced by ox-LDL

目的研究血管紧张素(Ang)(1-7)对氧化低密度脂蛋白(ox-LDL)诱导人脐静脉内皮细胞(HUVECs)内皮损伤、炎症反应的抑制作用。方法HUVECs细胞系经培养与传代后,分为对照组、模型组(ox-LDL诱导)、Ang(1-7)组、Ang(1-7)抑制剂(A779)组,采用MTT法检测各组细胞存活率,Annexin V-FITC/PI双染流式细胞术检测细胞凋亡情况,酶联免疫吸附法(ELISA)检测Ang(1-7)的含量和NO、eNOS水平及相关炎性因子TNF-α、IL-1β、IL-6的表达。结果ox-LDL成功诱导人脐静脉内皮细胞损伤模型。与对照组比较,模型组细胞存活率显著下降,凋亡率显著升高,Ang(1-7)、NO、eNOS水平显著降低,TNF-α、IL-1β、IL-6水平显著升高,差异均具有统计学意义(P<0.05);与模型组比较,Ang(1-7)组细胞存活率显著升高,细胞凋亡率显著下降,Ang(1-7)、NO、eNOS水平显著升高,TNF-α、IL-1β、IL-6水平显著下降,差异均具有统计学意义(P<0.05);与Ang(1-7)组比较,加入了Ang(1-7)抑制剂的A779组细胞存活率显著降低,细胞凋亡率显著升高,Ang(1-7)、NO、eNOS水平显著降低,TNF-α、IL-1β、IL-6水平显著升高,差异均具有统计学意义(P<0.05)。结论Ang(1-7)可改善ox-LDL诱导的人脐静脉内皮细胞存活率,降低其凋亡率,且通过调节eNOS活化,诱导NO释放,并减少TNF-α、IL-1β、IL-6等炎症因子的释放,降低ox-LDL诱导的人脐静脉内皮细胞损伤,具有抗动脉粥样硬化作用。

Objective To study the effect of Ang(1-7)on cell injury and inflammation of human umbilical vein endothelial cells(HUVECs)induced by oxidative low density lipoprotein(ox-LDL).Methods HUVECs were divided into four groups,including the control group,model group(ox-LDL induction),Ang(1-7)group and Ang(1-7)inhibitor(A779)group.The cell survival was measured by MTT method and cell apoptosis was detected by annexin V-FITC/PI double staining flow cytometry.Enzyme-linked immunosorbent assay(ELISA)was used to detect the levels of Ang(1-7),NO,eNOS,TNF-α,IL-1βand IL-6.Results The injury model of HUVECs was successfully induced by ox-LDL.Compared with the control group,the cell survival rate and the levels of Ang(1-7),NO and eNOS were markedly decreased,but the cell apoptosis rate and levels of TNF-α,IL-1βand IL-6 were significantly increased in the model group(P<0.05).Compared with the model group,Ang(1-7)treatment increased the cell survival rate and levels of Ang(1-7),NO and eNOS,but decreased the cell apoptosis rate and levels of TNF-α,IL-1βand IL-6(P<0.05).Compared with Ang(1-7)group,A779 treatment decreased the cell survival rate and the levels of Ang(1-7),NO and eNOS,but increased the levels of TNF-α,IL-1βand IL-6(P<0.05).Conclusion Ang(1-7)can improve the survival rate of ox-LDL induced HUVECs,and reduce the release of inflammatory factors such as TNF-α,IL-1βand IL-6,by regulation of eNOS activation and induction of NO release.Therefore,Ang(1-7)can reduce the injury of HUVECs induced by ox-LDL,which may be related to its anti-atherosclerotic activity.