Molecular mechanisms of Nrf2/Keap1 pathway and PINK/Parkin-mediated mitochondrial autophagy dysregulation in the progression of pre-eclampsia
Objective To explore the molecular mechanisms of Nrf2/Keap1 pathway and PINK/Parkin-mediated mitochondrial autophagy dysregulation in the progression of pre-eclampsia.Methods The endometrial samples derived from 15 pregnant women with a preterm birth history of severe pre-eclampsia(sPE)who underwent pregnancy examination and delivery in the Department of Obstetrics and Gynecology of Hainan Provincial People's Hospital during June 2020 to January 2023 were selected as the research subjects and included in sPE group.The endometrial samples derived from 15 pregnant women with a normal pregnancy and delivery history who underwent pregnancy examination and delivery in the same hospital during the same period were selected as control and included in nPCB group.Primary human endometrial stromal cells(hESCs)of endometrium in both groups were isolated.Reactive oxygen species(ROS)levels were determined by flow cytometry(FCM).Superoxide dismutase(SOD),reduced glutathione(GSH)and glutathione peroxidase(GPx)levels were determined by enzyme-linked immunosorbent assay(ELISA).hESCs were processed and grouped into nPCB-hESCs group,nPCB-hESCs treatment group,sPE-hESCs group,sPE-hESCs treatment group.The two sources of hESCs were treated with cAMP+MPA to induce decidualization in treatment groups.The expression levels of autophagy related proteins and mitochondrial autophagy related proteins were determined by Western blot.The implantation rate of JAr cells sphaeroplast into hESCs was determined.Another hESCs were selected for processing and grouping into nPCB-hESCs group,nPCB-hESCs+H/R group,sPE-hESCs group,sPE-hESCs+H/R group.Two resources of hESCs were treated with hypoxia/reoxygenation(H/R)in H/R groups.The mRNA and protein expression levels of Nrf2/Keap1 were determined by quantitative real-time polymerase chainreaction(qPCR)and Western blot.Results Compared with nPCB-hESCs,ROS levels in sPE-hESCs were significantly increased,while SOD,GSH and GPx levels were significantly decreased(P＜0.05).Compared with the nPCB-hESCs group,the expression levels of cellular autophagy/mitochondrial autophagy related proteins in nPCB-hESCs treatment group were significantly increased(P＜0.05).Compared with the nPCB-hESCs group,the implantation rate of JAr cells into hESCs was significantly lower in sPE-hESCs group(P＜0.05).Compared with the nPCB-hESCs treatment group,the implantation rate of JAr cells into hESCs was significantly lower in sPE-hESCs treatment group(P＜0.05).Compared with nPCB-hESCs+H/R group,the mRNA and protein expression levels of Nrf2 in sPE-hESCs+H/R group were significantly decreased,and the mRNA and protein expression levels of Keap1 were significantly increased(P＜0.05).Conclusions The Nrf2/Keap1 pathway activation is impaired and the mitochondrial autophagy function is inhibited in hESCs of individuals with pre-eclampsia history,leading to the abnormal accumulation of intracellular ROS and the loss of decidualization function in vitro.
Pre-eclampsiaPrimary human endometrial stromal cellsOxidative stressMitochondrial autophagyNrf2/Keap1
万方数据
维普
