Involvement of phosphatase and tensin homolog-induced putative kinase 1–Parkin-mediated mitophagy in septic acute kidney injury  

作　　者：Xin-Gui Dai Wei Xu Tao Li Jia-Ying Lu Yang Yang Qiong Li Zhen-Hua Zeng Yu-Hang Ai 

机构地区：[1]Department of Intensive Care Unit,Xiangya Hospital,Central South University,Changsha,Hunan 410078,China [2]Department of Critical Care Medicine,The First People's Hospital of Chenzhou,Chenzhou,Hunan 423000,China [3]Department of Critical Care Medicine,Nanfang Hospital,Southern Medical University,Guangzhou,Guangdong 510515,China

出　　处：《Chinese Medical Journal》2019年第19期2340-2347,共8页中华医学杂志（英文版）

基　　金：This work was supported by grants from the National Natural Science Foundation of China(Nos.81601708,81671960);the Natural Science Foundation of Hunan Province,China(No.2018JJ2014).

摘　　要：Background:Studies have reported mitophagy activation in renal tubular epithelial cells(RTECs)in acute kidney injury(AKI).Phosphatase and tensin homolog-induced putative kinase 1(PINK1)and E3 ubiquitin-protein ligase Parkin are involved in mitophagy regulation;however,little is known about the role of PINK1-Parkin mitophagy in septic AKI.Here we investigated whether the PINK1-Parkin mitophagy pathway is involved in septic AKI and its effects on cell apoptosis in vitro and on renal functions in vivo.Methods:Mitophagy-related gene expression was determined using Western blot assay in human RTEC cell line HK-2 stimulated with bacterial lipopolysaccharide(LPS)and in RTECs from septic AKI rats induced by cecal ligation and perforation(CLP).Autophagy-related ultrastructural features in rat RTECs were observed using electron microscopy.Gain-and loss-of-function approaches were performed to investigate the role of the PINK1-Parkin pathway in HK-2 cell mitophagy.Autophagy activators and inhibitors were used to assess the effects of mitophagy modulation on cell apoptosis in vitro and on renal functions in vivo.Results:LPS stimulation could significantly induce LC3-II and BECN-1 protein expression(LC3-II:1.72±0.05 vs.1.00±0.05,P<0.05;BECN-1:5.33±0.57 vs.1.00±0.14,P<0.05)at 4 h in vitro.Similarly,LC3-II,and BECN-1 protein levels were significantly increased and peaked at 2 h after CLP(LC3-II:3.33±0.12 vs.1.03±0.15,P<0.05;BECN-1:1.57±0.26 vs.1.02±0.11,P<0.05)in vivo compared with those after sham operation.Mitochondrial deformation and mitolysosome-mediated mitochondria clearance were observed in RTECs from septic rats.PINK1 knockdown significantly attenuated LC3-II protein expression(1.35±0.21 vs.2.38±0.22,P<0.05),whereas PINK1 overexpression markedly enhanced LC3-II protein expression(2.07±0.21 vs.1.29±0.19,P<0.05)compared with LPS-stimulated HK-2 cells.LPS-induced proapoptotic protein expression remained unchanged in autophagy activator-treated HK-2 cells and was significantly attenuated in PINK1-overexpressing

关 键 词：Sepsis Acute kidney injury Autophagy MITOPHAGY PHOSPHATASE and TENSIN homolog-induced putative KINASE 1 E3 ubiquitin-protein ligase Parkin 

分 类 号：R69[医药卫生—泌尿科学]