Nrf2/Keap1通路和PINK/Parkin介导的线粒体自噬失调在子痫前期进展中的分子机制研究  

Molecular mechanisms of Nrf2/Keap1 pathway and PINK/Parkin-mediated mitochondrial autophagy dysregulation in the progression of pre-eclampsia

作　　者：王莉[1] 孔娇 林青[1] 郑林媚[1] 郑颖 王艾桦 陈小菊[1] WANG Li;KONG Jiao;LIN Qing;ZHENG Linmei;ZHENG Ying;WANG Aihua;CHEN Xiaoju(Obstetrics Zone 1,Hainan Provincial People's Hospital(Hainan Hospital Affiliated to Hainan Medical College),Haikou 570013,Hainan,China;Pathology Department,Hainan Provincial People's Hospital(Hainan Hospital Affiliated to Hainan Medical College),Haikou 570013,Hainan,China)

机构地区：[1]海南省人民医院(海南医学院附属海南医院)产科一区,海口570013 [2]海南省人民医院(海南医学院附属海南医院)病理科,海口570013

出　　处：《中国性科学》2024年第3期104-109,共6页Chinese Journal of Human Sexuality

基　　金：海南省自然科学基金高层次人才项目(822RC810)。

摘　　要：目的探讨Nrf2/Keap1通路和PINK/Parkin介导的线粒体自噬失调在子痫前期疾病进展中的分子机制。方法选取2020年6月至2023年1月于海南省人民医院妇产科做孕期检查并分娩的15例有重度子痫前期(sPE)早产史的孕妇的子宫内膜样本作为研究对象,纳入sPE组;并选取同期同院做孕期检查并分娩的15例有正常孕产史的孕妇的子宫内膜样本作为对照,纳入nPCB组。分离两组原代子宫内膜基质细胞(hESCs)。采用流式细胞术(FCM)测定活性氧(ROS)水平,采用酶联免疫吸附试验(ELISA)法测定总超氧化物歧化酶(SOD)、谷胱甘肽(GSH)和谷胱甘肽过氧化物酶(GPx)水平。将hESCs进行处理并分组为nPCB-hESCs组、nPCB-hESCs处理组、sPE-hESCs组、sPE-hESCs处理组。分别对处理组两种来源的hESCs给予cAMP+MPA诱导蜕膜化。采用Western blot测定自噬相关蛋白和线粒体自噬相关蛋白的表达水平,测定JAr细胞球状体向hESCs的植入率。另选取hESCs进行处理并分组为nPCB-hESCs组、nPCB-hESCs+H/R组、sPE-hESCs组、sPE-hESCs+H/R组。H/R组分别对两种来源的hESCs给予缺氧/复氧(H/R)处理。采用实时荧光定量聚合酶链反应(qPCR)和Western blot测定Nrf2/Keap1 mRNA和蛋白质的表达水平。结果与nPCB-hESCs相比,sPE-hESCs中ROS的水平显著升高,SOD、GSH和GPx的水平显著降低(P<0.05)。与nPCB-hESCs组相比,nPCB-hESCs处理组细胞自噬/线粒体自噬相关蛋白表达水平显著升高(P<0.05)。与nPCB-hESCs组相比,sPE-hESCs组JAr细胞向hESCs植入的植入率显著较低(P<0.05)。与nPCB-hESCs处理组相比,sPE-hESCs处理组JAr细胞向hESCs植入的植入率显著较低(P<0.05)。与nPCB-hESCs+H/R组相比,sPE-hESCs+H/R组细胞内Nrf2 mRNA和蛋白质的表达水平显著降低,Keap1 mRNA和蛋白质的表达水平显著升高(P<0.05)。结论有子痫前期史的孕妇hESCs中Nrf2/Keap1通路激活受损和线粒体自噬功能被抑制,导致细胞内ROS的异常积累及其体外蜕膜化功能受�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,t

关 键 词：子痫前期 原代人子宫内膜基质细胞 氧化应激 线粒体自噬 Nrf2/Keap1 

分 类 号：R714[医药卫生—妇产科学]