GLS1通过激活Nrf2/HO-1轴抑制过氧化氢诱导的ARPE-19细胞氧化应激、自噬与凋亡  

GLS1 inhibits hydrogen peroxide-induced oxidative stress,autophagy and apoptosis in ARPE-19 cells by activating the Nrf2/HO-1

作　　者：周洋[1] 美丽巴努·玉素甫[1] 陈婷妍 ZHOU Yang;Meilibanu·Yusufu;CHEN Tingyan(Department of Ophthalmology,the Fifth Affiliated Hospital of Xinjiang Medical University,Xinjiang,Urumqi 830000,China;不详)

机构地区：[1]新疆医科大学第五附属医院眼科,乌鲁木齐市830000 [2]成都华厦眼科医院小儿斜弱视科

出　　处：《河北医药》2023年第19期2901-2905,共5页Hebei Medical Journal

基　　金：新疆维吾尔自治区自然科学基金项目(编号:2019D01C271)。

摘　　要：目的探究GLS1对过氧化氢诱导的ARPE-19细胞氧化应激、自噬与凋亡的影响及机制。方法ARPE-19细胞分为对照组、H_(2)O_(2)组、H_(2)O_(2)+Vector组、H_(2)O_(2)+GLS1组,对照组细胞不做任何处理,H_(2)O_(2)组细胞用200μmol/L H_(2)O_(2)诱导48h,H_(2)O_(2)+Vector组和H_(2)O_(2)+GLS1组细胞转染Vector和GLS1质粒后用200μmol/L H_(2)O_(2)诱导48 h,比色法测定各组细胞SOD、GSH和MDA浓度,透射电镜观察各组细胞自噬小体,流式细胞术检测各组细胞凋亡水平,Western blot检测各组细胞Nrf2、HO-1、LC3-Ⅱ、p62的表达。结果与对照组比较,H_(2)O_(2)组ARPE-19细胞SOD、GSH表达显著下降,MDA显著增加,自噬小体数目显著增加,细胞凋亡显著增加,LC3-Ⅱ表达显著上调,P62、抗核因子红系2相关因子2(nuclear factor erythroid 2-related factor 2,Nrf2)、血红素氧合酶1(heme Oxygenase-1,HO-1)表达显著下调。与H_(2)O_(2)+Vector组比较,H_(2)O_(2)+GLS1组细胞SOD、GSH表达显著增加,MDA显著降低,自噬小体数目显著减少,细胞凋亡显著减少,LC3-Ⅱ表达显著下调,P62、Nrf2、HO-1表达显著上调。结论GLS1可抑制H_(2)O_(2)诱导的ARPE-19细胞氧化应激、自噬与凋亡,其机制为激活Nrf2/HO-1信号通路。Objective To explore the effect of glutaminase 1(GLS1)on hydrogen peroxide(H_(2)O_(2))-induced oxidative stress,autophagy and apoptosis in human retinal pigment epithelial(ARPE-19)cells,and the underlying mechanism.Methods ARPE-19 cells were divided into control group(blank control),H_(2)O_(2)group(200μmol/L H_(2)O_(2)induction for 48h),H_(2)O_(2)+Vector group(transfection of vector,followed by 200μmol/L H_(2)O_(2)induction for 48h)and H2O2+GLS1 group(transfection of GLS1 vector,followed by 200μmol/L H_(2)O_(2)induction for 48h).The concentrations of superoxide dismutase(SOD),glutathione(GSH)and malondialdehyde(MDA)in each group were determined by colorimetry.Autophagosomes in each group were observed by transmission electron microscopy,and the level of apoptosis in each group was detected by flow cytometry.Western blot was used to detect the protein expressions of Nrf2(nuclear factor erythroid 2-related factor 2),HO-1(heme oxygenase-1),LC3-Ⅱand p62 in each group of cells.Results Compared with the control group,ARPE-19 cells in the H_(2)O_(2)group presented significantly lower SOD and GSH levels,higher MDA level,higher number of autophagosomes,higher apoptotic rate,upregulation of LC3Ⅱ,and downregulation of p62,Nrf2 and HO-1.The opposite trends of the above-mentioned indexes were detected in H_(2)O_(2)+GLS1 group than those in H_(2)O_(2)+Vector group.Conclusion GLS1 can inhibit H_(2)O_(2)-induced oxidative stress,autophagy and apoptosis in ARPE-19 cells by activating the Nrf2/HO-1 signaling pathway.

关 键 词：GLS1 H2O2诱导 ARPE-19细胞 Nrf2/HO-1轴 

分 类 号：R774.5[医药卫生—眼科]