NaAsO2对L-02肝细胞氧化应激性损伤及SOD1、AUF1表达影响  

Effects of NaAaO2 on oxidative stress injury and expression of SOD1 and AUF1 in L-02 hepatocytes

作　　者：胡倩 时明阳 毕顶念 李娇 吕莹 段恬筱 付雨 张爱华 胡勇 HU Qian;SHI Ming-yang;BI Ding-nian;LI Jiao;LV Ying;DUAN Tian-xiao;FU Yu;ZHANG Ai-hua;HU Yong(Key Laboratory of Enironmental Pllution Monitoring and Disease Control,Ministry of Education,School of Public Health,Cuichou Medical Unirersity,Guiyang,Guichou 550025,China)

机构地区：[1]贵州医科大学环境污染与疾病监控教育部重点实验室,贵州医科大学公共卫生学院,贵州贵阳550025

出　　处：《现代预防医学》2020年第23期4321-4325,共5页Modern Preventive Medicine

基　　金：国家自然科学基金(81860561);贵州省科学技术基金(黔科合基础[2016]1119)。

摘　　要：目的观察不同剂量亚砷酸钠(NaAsO2)对L-02肝细胞氧化应激性损伤作用及超氧化物歧化酶1(SOD1)、AU碱基富集元件RNA结合因子1(AUF1)表达影响。方法用0、10、20、40μmol/L NaAsO2分别处理L-02肝细胞24小时,化学比色法检测谷胱甘肽巯基转移酶(GST)、γ-谷氨酰转肽酶(γ-GT)活力及总胆汁酸(TBA)含量和SOD1、谷胱甘肽过氧化物酶(GPx)活性;荧光探针2,7-二氯二氢荧光素二乙酸酯检测细胞内活性氧族(ROS)相对荧光密度;实时荧光定量PCR和Western Blotting分别检测SOD1和AUF1转录和蛋白表达。结果 (1)与对照组比较,20、40μmol/L NaAsO2组GST活性和TBA含量均升高(P<0.05);10、20、40μmol/L NaAsO2组γ-GT活性也升高(P<0.05)。(2)与对照组比较,20、40μmol/L NaAsO2组SOD1活性均下降(P<0.05);GPx活性仅在10μmol/L升高(P<0.05);细胞内ROS先降低后升高(P<0.05)。(3)与对照组比较,20和40μmol/L NaAsO2组AUF1和SOD1 mRNA表达均升高(P<0.05),AUF1蛋白表达先升高后降低(P<0.05);10、20μmol/L NaAsO2组SOD1蛋白表达升高(P<0.05),但40μmol/L NaAsO2组SOD1蛋白表达有降低趋势。结论 NaAsO2对L-02肝细胞产生氧化应激性损伤,其机制可能是NaAsO2通过降低AUF1的蛋白表达降低,从转录后调控降低了SOD1 mRNA的稳定性而使其蛋白表达和酶活力降低。Objective To observe effects of sodium arsenite(NaAsO2)at different concentrations on oxidative stress injury and the expression of superoxide dismutase 1(SOD1)and AU-rich element RNA-binding factor 1(AUF1)in L-02 hepatocytes.Methods The L-02 hepatocytes were exposed to 0,10,20,40μmol/L NaAsO2 for 24 hours,and then the colorimetry was used to detect levels of glutathione S-transferase(GST),γ-glutamyltranspeptidase(γ-GT),total bile acid(TBA)and SOD1,glutathione peroxidase(GPx).Fluorescent probe 2’,7’-dichlorodihydrofluorescein diacetate was used to detect relative fluorescence density of reactive oxygen species(ROS)in cells.Real-time quantitative PCR and Western blotting were used to detect the transcription and protein expression of SOD1 and AUF1,respectively.Results(1)With increasing of NaAsO2 concentrations,the levels of GST and TBA in 20 and 40μmol/L NaAsO2 groups increased more than those in the control group(P<0.05),theγ-GT activity in 10,20 and 40μmol/L NaAsO2 groups also increased(P<0.05).(2)With the increasing of NaAsO2 concentrations,compared with the control group,the SOD1 activity in the 20 and 40μmol/L NaAsO2 groups decreased(P<0.05).However,the GPx activity increased only in 10μmol/L group compared with that in the control group(P<0.05).The relative fluorescence density of ROS decreased at first and then increased(P<0.05).(3)With increasing of NaAsO2 concentrations,the mRNA expression levels of AUF1 and SOD1 in 20 and 40μmol/L NaAsO2 groups increased than those in the control group(P<0.05);the protein expression levels of AUF1 increased firstly and decreased at last(P<0.05);the protein expression levels of SOD1 in the 10 and 20μmol/L NaAsO2 groups increased than those in the control group(P<0.05),but there only had a decreasing trend in the 40μmol/L NaAsO2 group.Conclusion NaAsO2 exposure could cause oxidative stress damage to L-02 hepatocytes.The mechanism may be that NaAsO2 could reduce the AUF1 protein expression.That cause the stability of SOD1 mRNA reduction by post-transcriptiona

关 键 词：亚砷酸钠 SOD1 AUF1 

分 类 号：R114[医药卫生—卫生毒理学]