Protective effect of UCP2 on ANGII-induced mitochondrial damage in endothelial cells
Objective To investigate the protective effect and molecular mechanism of mitochondrial uncoupling protein 2(UCP2)on mitochondrial dysfunction induced by angiotensin Ⅱ(ANG Ⅱ)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 blot-ting thed using COX Ⅳ as 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-regulating UCP2 expression in HUVECs,Western-blotting combined with Real-time PCR was used to detect changes in mito-chondrial damage-related proteins and gene expression levels.Results ANG Ⅱ inhibited the proliferation of HUVECs in vitro,and showed a certain concentration dependence.As the concentration of ANGⅡ increased,Western-blotting detection showed that the ex-pression 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 in-creased(P<0.05).Conclusion ANGⅡ can 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.
NETL
NSTL
万方数据
