Nrf2基因的过表达对酒精诱导的HepG2细胞损伤的保护作用及机制  

Overexpression of Nrf2 effectively ameliorates oxidative stress and inflammation induced by ethanol in HepG2 cell line

作　　者：朱炜[1] 俞宏斌[1] 戴闯[1] 冯文明[1] 

机构地区：[1]浙江省湖州市第一人民医院分子外科研究所

出　　处：《中国临床药理学与治疗学》2016年第1期55-59,共5页Chinese Journal of Clinical Pharmacology and Therapeutics

摘　　要：目的:探讨于人类肝癌细胞系HepG2中过表达Nrf2基因后对由酒精处理引起的细胞氧化压力和炎症反应的影响和机制。方法:经过Keap1 siRNA预处理的HepG2细胞经含75 mmol/L酒精的培养基孵育72 h后,应用四甲基偶氮唑盐比色法(MTT法)检测细胞活性以及应用Western blot和荧光定量实时聚合酶链反应(PCR)方法检测细胞内氧化压力和促炎症蛋白的表达水平。同时,转录因子核因子κB(NF-κB)的活性以及其抑制蛋白IκBα的表达也通过Western blot检测。结果:经含75 mmol/L酒精的培养基孵育72 h后,HepG2细胞的细胞活性显著下降,同时伴随着细胞内抗氧化酶表达水平的显著下降和促炎症蛋白表达的显著上升。Nrf2基因的过表达显著增加了HepG2细胞活性,恢复了细胞内抗氧化酶的表达水平,并改善了细胞内的炎症反应(P<0.05)。Nrf2的过表达亦通过恢复IκBα的水平降低了由酒精处理诱导的转录因子NF-κB的活性(P<0.05)。结论:Nrf2基因的过表达可以有效改善酒精造成的HepG2细胞内氧化压力和炎症反应,该效应部分通过抑制NF-κB的活性来完成。AIM： To study the effect and mechanism of Nrf2 overexpression on ethanol-induced oxidative stress and inflammation in HepG2 cell line.METHODS： HepG2 cells pre-treated with Keap1 si RNA was incubated in medium containing 75 mmol/L ethanol for 72 hours.After that,cell viability was measured by MTT assay and key markers for oxidative stress and inflammation were measured by quantitative realtime PCR or Western blot.In addition,the activity of NF-κB and the expression of its inhibitory protein IκBα were examined by Western blot.RESULTS： After 72-hour incubation with medium containing 75 mmol/L ethanol,HepG2 cells showed decreased viability,with reduced antioxidant enzymatic level and increased inflammatory marker level.Overexpression of Nrf2 significantly enhanced the viability of HepG2 cell,restored the expression level of antioxidant enzymes,and improved inflammation（P〈0.05）.Silence of Nrf2 also reduced the activity of NF-κB via restoring the level of IκBα.CONCLUSION： Overexpression of Nrf2 in HepG2 cell effectively ameliorated oxidative stress and inflammation induced by ethanol incubation partly through inhibiting the activity of NF-κB.

关 键 词：Nrf2 KEAP1 酒精 HEPG2细胞 

分 类 号：R994.4[医药卫生—毒理学]