UCP2对ANG Ⅱ诱导的内皮细胞线粒体损伤的保护作用  

Protective effect of UCP2 on ANGII-induced mitochondrial damage in endothelial cells

作　　者：陈鹏[1] 杨红年[1] 杜立文[1] 关海昕[1] 崔彬 张玲[1] CHEN Peng;YANG Hongnian;DU Liwen;GUAN Haixin;CUI Bin;ZHANG Ling(Department of Emergency,People′s Hospital of Ningxia Hui Autonomous Region,Yinchuan 750002,China)

机构地区：[1]宁夏回族自治区人民医院急诊科,宁夏银川750002

出　　处：《宁夏医学杂志》2024年第7期565-568,I0001,共5页Ningxia Medical Journal

基　　金：宁夏自然科学基金项目(2021AAC03306)。

摘　　要：目的探究线粒体解偶联蛋白2(UCP2)对由血管紧张素Ⅱ(ANGⅡ)刺激引发的人脐静脉内皮细胞(HUVECs)线粒体功能损害的保护作用及分子机制。方法培养HUVECs,给予ANGⅡ处理24 h,CCK8实验检测细胞活性变化;Western-blotting实验以COXⅣ为内参蛋白,检测线粒体损伤相关蛋白MDM2和ATM及UCP2表达水平变化;Real-time PCR实验以GAPDH为内参基因,检测线粒体损伤基因ATM、ENOS、MDM2和MNSOD及UCP2表达水平变化,评价细胞内氧化状态;进一步通过上调和下调HUVECs中UCP2表达,Western-blotting联合Real-time PCR检测线粒体损伤相关蛋白及基因表达水平变化。结果ANGⅡ能够抑制HUVECs增殖,同时表现出一定浓度依赖性。随着ANGⅡ浓度升高,Western-blotting检测发现ATM表达降低、UCP2和MDM2表达增高(P<0.05);Real-time PCR检测发现ATM和ENOS表达降低,MDM2、UCP2和MNSOD表达增高(P<0.05)。过表达UCP2慢病毒感染细胞后,Western-blotting检测发现MDM2及ATM表达降低,Real-time PCR检测发现ATM、MDM2及MNSOD表达降低,UCP2敲低质粒转染细胞后,Western-blotting检测发现MDM2及ATM表达增高,Real-time PCR检测发现ATM、MDM2及MNSOD表达增高(P<0.05)。结论ANGⅡ可引起HUVECs线粒体损伤,且UCP2在这一过程中发挥保护作用,可减轻ANGⅡ导致的HUVECs线粒体损伤。Objective To investigate the protective effect and molecular mechanism of mitochondrial uncoupling protein 2(UCP2)on mitochondrial dysfunction induced by angiotensinⅡ(ANGⅡ)in human umbilical vein endothelial cells(HUVECs).Methods HUVECs was cultured,and then exposure to ANGⅡfor 24-hour.CCK8 was used to test the cell activity and Western blotting thed using COXⅣas the internal reference protein was used for evaluating mitochondrial injury markers MDM2,ATM,and UCP2.Real-time PCR was used to detect the expression levels of mitochondrial damage genes ATM,ENOS,MDM2,MNSOD and UCP2,with GAPDH as the internal reference gene,and the intracellular oxidation state was also evaluated.Furthermore,by up-regulating and down-reg ulating UCP2 expression in HUVECs,Western-blotting combined with Real-time PCR was used to detect changes in mitochondrial damage-related proteins and gene expression levels.Results ANGⅡinhibited the proliferation of HUVECs in vitro,and showed a certain concentration dependence.As the concentration of ANGⅡincreased,Western-blotting detection showed that the expression of ATM decreased and the expression of UCP2 and MDM2 increased(P<0.05).Real-time PCR showed that the expression of ATM,ENOS was decreased,and the expression of MDM2 UCP2 and MNSOD was increased(P<0.05).After overexpression of UCP2 lentivirus,Western-blotting showed that the expression of MDM2 and ATM was decreased.Real-time PCR showed that the expression of ATM,MDM2 and MNSOD was decreased.After UCP2 knockdown plasmid was transfected into cells,Western-blotting showed that the expression of MDM2 and ATM was increased.Real-time PCR showed that the expression of ATM,MDM2 and MNSOD was increased(P<0.05).Conclusion ANGⅡcan cause mitochondrial damage in HUVECs,and UCP2 plays a protective role in this process and it can reduce the mitochondrial damage in HUVECs caused by ANGⅡ.

关 键 词：HUVECS 线粒体损伤 血管紧张素Ⅱ 线粒体解偶联蛋白2 

分 类 号：R34[医药卫生—基础医学]