白藜芦醇对酒精诱导的HepG2细胞氧化损伤的保护作用及相关基因表达的变化  

Resveratrol protects against alcohol-induced oxidative stress in human HepG2 cells

作　　者：何培元[1] 高淑梅[2] 侯志平[3] 马立新[1] 李炳庆[1] 

机构地区：[1]承德医学院附属医院消化内科,河北省承德市067000 [2]承德医学院附属医院心电室,河北省承德市067000 [3]承德医学院病理教研室,河北省承德市067000

出　　处：《世界华人消化杂志》2014年第14期1928-1935,共8页World Chinese Journal of Digestology

基　　金：河北省卫生厅基金资助项目;No.20110179~~

摘　　要：目的:探讨白藜芦醇在酒精诱导的HepG2细胞氧化应激中的抗氧化作用,揭示白藜芦醇抗酒精性肝损伤的作用机制.方法:白藜芦醇预处理HepG2细胞24 h后,用酒精诱导氧化应激的产生.MTT方法检测白藜芦醇处理组与对照组HepG2细胞活力;用ELISA试剂盒检测不同实验组的超氧化物歧化酶(superoxide dismutase,SOD)和细胞内总活性氧(reactive oxygen species,ROS)含量;采用RTPCR方法检测抗氧化通路中关键基因SOD1、SOD2和过氧化氢酶的mRNA表达水平.结果:MTT结果显示,与对照组比较,白藜芦醇(12.5、25、50、100μmol/L)对HepG2细胞的细胞毒性均在20%以下,300 mmol/L酒精可致近50%HepG2细胞死亡;25-100μmol/L白藜芦醇可有效对抗300 mmol/L酒精对HepG2引起的细胞毒性作用;SOD活性显示100μmol/L白藜芦醇预处理组SOD含量(0.391±0.011)明显高于非处理组(0.286±0.019),而ROS结果显示酒精诱导组(29234.79±2288)明显高于白藜芦醇25、50、100μmol/L预处理组(18023.26±1359.66;13528.44±1078.99;8219.87±635.99);RT-PCR结果显示,与300mmol/L酒精比较,25、50和100μmol/L白藜芦醇可上调SOD1 mRNA表达量(0.5535±0.0035;0.586±0.0113;0.623±0.0127);12.5、25、50、100μmol/L白藜芦醇均可上调SOD2mRNA表达量(1.249±0.011;1.369±0.028;1.377±0.021;1.401±0.0578);50和100μmol/L白藜芦醇均可上调过氧化氢酶(0.1955±0.004;0.2275±0.00707)mRNA的表达量.结论:本研究提示酒精可诱导氧化应激的产生,而白藜芦醇通过调节抗氧化通路中基因的表达发挥抗氧化功能,从而削弱酒精的细胞损伤作用.AIM： To assess the effect of resveratrol as an antioxidant on alcohol-induced oxidative stress in HepG2 cells and to explore the possible mechanisms involved. METHODS： HepG2 cells were pretreated with resveratrol for 24 h before treatment with alcohol to induce oxidative stress. MTT assay was performed to detect the viability of resveratroltreated HepG2 cells and control cells. ELISA was used to detect the activity of superoxide dismutase （SOD） and the level of total intracellular reactive oxygen species （ROS）. Finally, RT-PCR was performed to detect the expression levels of SOD1, SOD2 and catalase, which are key genes involved in anti-oxidation pathway. RESULTS： In comparison with control cells （not treated with resveratrol）, the toxicity of resveratrol （12.5, 25, 50, 100 μmol/L） towards HepG2 cells during pre-treatment was below 20%. Treatment with 300 mmol/L alcohol without pre- treatment with resveratrol resulted in the death of around 50% of cells. Resveratrol at concentrations ranging from 25 to 100 μmol/L had an antagonistic effect against cytotoxicity of 300 mmol/L alcohol to HepG2 cells. SOD activity was significantly higher in cells pre-treated with 100 mmol/L resveratrol （0.391 ± 0.011） compared to non-treated controls （0.391 ± 0.011 vs 0.286± 0.019, P 〈 0.05）. After alcohol induction, non-treated cells showed a higher ROS level compared to cells treated with 25, 50, and 100 ~mol/L resveratrol （29234.79 ± 2288.00 vs 18023.26 ± 1359.66, 13528.44 ± 1078.99, 8219.87 ± 635.99）. Compared to cells induced with 300 mM alcohol resveratrol increased the expres- sion levels of SOD1 （0.5535± 0.0035, 0.586 ± 0.0113, 0.623 ± 0.0127）, SOD2 （1.249± 0.011, 1.369± 0.028, 1.377 ± 0.021, 1.401 ± 0.0578） and catalase （0.1955 ± 0.004, 0.2275 ±0.00707）.CONCLUSION： Resveratrol as an antioxidant protects against alcohol-induced oxidative stress by regulating the expression of genes involved in anti-oxidant pathways.

关 键 词：白藜芦醇 氧化应激 HEPG2 抗氧化 

分 类 号：R575.5[医药卫生—消化系统]