Mechanism of RvE1 inhibiting hypoxic endothelial cell injury by regulating Golgi stress
Objective To explore the role of Golgi stress in the injury of hypoxic vascular endothelial cells,and to explore the regulatory effect of fading hormone E1(RvE1)on Golgi stress.Methods Human umbilical vein endothelial cells were divided into 20.9％O2,2％O2,2％O2+si-NC,2％O2+si-GRASP65,2％O2+RvE1,2％O2+RvE1+OE-GRASP65-NC and 2％O2+RvE1+OE-GRASP65 groups.The morphological changes of HUVEC Golgi apparatus were observed by transmission electron microscopy,cell viability was detected by CCK-8,reactive oxygen species(ROS)levels were detected by flow cytometry,and the changes of endothelin-1(ET-1),prostacyclin-2(PGI-2)and RvE1 were detected by ELISA.Real-time quantitative fluorescent PCR(qRT-PCR)and Western blotting were used to detect the mRNA and protein expression of nitric oxide synthetase(eNOS)and Golgi in vitro peripheral membrane protein P65 antibody(GRASP65).Results Compared with 20.9％O2 group,hypoxia could cause rupture or even remodeling of the Golgi apparatus of vascular endothelial cells,significantly inhibit HUVEC activity(P＜0.01),significantly up-regulate ROS production(P＜0.01),and significantly inhibit the expressions of eNOS,PGI-2 and RvE1(P＜0.01).Significantly promoted the expression of ET-1,GRASP65,p-GRASP65(P＜0.01);Compared with 2％O2 group,si-GRASP65 and RvE1 could improve the stress response of hypoxic HUVEC Golgi apparatus,significantly enhance HUVEC activity(P＜0.01),significantly down-regulate ROS production(P＜0.01),and significantly increase the expressions of eNOS and PGI-2(P＜0.01).The expression of ET-1,GRASP65 and p-GRASP65 was significantly inhibited(P＜0.01).OE-GRASP65 has a significant cancelling effect on the efficacy of RvE1.Conclusions RvE1 can antagonize hypoxic injury of endothelial cells possibly by inhibiting hypoxia-induced Golgi stress.
resolvin E1hypoxiaendothelial cellsGolgi stressreactive oxygen species
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