萝卜硫素对H2O2诱导人肝细胞氧化应激损伤模型的保护作用  

Protective effects of sulforaphane in a H2O2 induced oxidative stress model of human hepatocytes

作　　者：谭静 易国栋 Tan Jing;Yi Guodong(Department of Nephrorheumatology,Central Hospital of Enshi Autonomous Prefecture,Enshi 445000,Hubei Province,China;Department of Gastroenterology,Central Hospital of Enshi Autonomous Prefecture,Enshi 445000,Hubei Province,China)

机构地区：[1]恩施土家族苗族自治州中心医院肾病风湿科,湖北省恩施市445000 [2]恩施土家族苗族自治州中心医院消化内科,湖北省恩施市445000

出　　处：《中国组织工程研究》2020年第31期5052-5056,共5页Chinese Journal of Tissue Engineering Research

摘　　要：背景:氧化应激损伤在慢性肝炎发生发展中起着重要的作用,而天然产物萝卜硫素具有较好的抗氧化活性。目的:研究萝卜硫素对H2O2诱导人肝细胞(LO2)氧化应激损伤的保护作用,并探讨其可能的作用机制。方法:体外培养LO2细胞,采用H2O2诱导LO2细胞造模,并同时将细胞分为对照组、H2O2诱导组、H2O2+低剂量萝卜硫素组(10μmol/L)、H2O2+中剂量萝卜硫素组(20μmol/L)及H2O2+高剂量萝卜硫素组(40μmol/L)。采用100μg/L的H2O2处理人肝细胞24 h,诱导细胞氧化应激损伤;H2O2+萝卜硫素组将H2O2与不同浓度萝卜硫素同时处理细胞24 h。采用CCK8法分析萝卜硫素对细胞存活率的影响,分析细胞匀浆中的氧化应激产物水平,Western Blot法分析Nrf2、GCLC、NQO1及血红素加氧酶1蛋白表达水平。研究方案的实施符合恩施土家族苗族自治州中心医院的相关伦理要求。结果与结论:①H2O2能诱导LO2细胞氧化应激损伤;②相比于H2O2模型组,各浓度萝卜硫素能诱导LO2细胞增殖、抑制丙二醛和一氧化氮的产生及促使超氧化物歧化酶活性升高(P<0.05);③Western Blot结果显示萝卜硫素能诱导Nrf2向细胞核内转移,诱导GCLC、NQO1和血红素加氧酶1蛋白表达(P<0.05);④结果说明,萝卜硫素能改善H2O2诱导的LO2细胞氧化应激损伤,其药理学作用与Nrf2信号传导活化有关。BACKGROUND:Oxidative stress injury plays an important role in the development of chronic hepatitis,and sulforaphane is a natural product with good antioxidant activity.OBJECTIVE:To investigate the protective effects of sulforaphane on oxidative stress injury of human hepatocytes(LO2)stimulated by H2O2,and to investigate its possible mechanism.METHODS:LO2 cells were cultured in vitro,followed by 24-hour treatment with 100μg/L H2O2 to induce the oxidative stress model of the cells.The cells were divided into five groups:control group,model group,sulforaphane treatment groups(low-,middle-,high-dose groups:H2O2+10,20,and 40μmol/L sulforaphane,respectively).Cell counting kit-8 was used to measure the relative survival rate of cells,and analyze the levels of oxidative stress products in cell homogenate.Western blot assay was used to determine the expression levels of nuclear factor erythroid-2 related factor 2(Nrf2),GCLC,NQO1 and heme oxygenase-1.The study protocol was in accordance with the related ethic requirements of Central Hospital of Enshi Autonomous Prefecture.RESULTS AND CONCLUSION:H2O2 could induce oxidative stress injury in LO2 cells.Compared with the model group,sulforaphane at various concentrations could significantly induce the proliferation of LO2 cells,inhibit the production of malondialdehyde and nitric oxide,and promote the activity of superoxide dismutase(P<0.05).Western blot results revealed that sulforaphane could induce the transfer of Nrf2 into the nucleus and significantly increase the expression of GCLC,NQO1,and heme oxygenase-1 proteins(P<0.05).To conclude,sulforaphane can significantly relieve the oxidative stress injury of LO2 cells induced by H2O2 and its underlying mechanism may be related to the activation of Nrf2 pathway.

关 键 词：萝卜硫素 人肝细胞 氧化应激 血红素加氧酶1 核因子E2相关因子2 蛋白 

分 类 号：R446[医药卫生—诊断学] R496[医药卫生—临床医学]