摘要
目的:研究胡黄连苷Ⅱ(PicrosideⅡ,PicⅡ)对过氧化氢(H2O2)诱导的人肝细胞系L-02氧化损伤中转录因子NF-E2相关因子(Nrf2)及下游Ⅱ相抗氧化解毒酶表达的影响,探讨胡黄连苷Ⅱ抗氧化保护肝细胞的作用机制。方法:用0.5 mmol/L,5mmol/L picⅡ干预L-02细胞3 h作为干预组,用DMEM培养作为对照及模型组,除对照组外各组均加入终浓度为0.6 mmol/L的H2O2作用1 h造成急性氧化应激损伤,后继续培养24 h,MTT法检测细胞增殖状况,速率法检测细胞培养液AST和ALT的水平,通过实时荧光定量RT-PCR检测各组的Nrf2、NQO1、GST、Gclc和Gclm的mRNA的表达变化。结果:胡黄连苷Ⅱ可降低由H2O2诱导的L-02细胞损伤(P<0.01),模型组AST、ALT水平较对照组显著升高(P<0.01),胡黄连苷Ⅱ干预组较模型组均降低(P<0.01);模型组NQO1、Gclc、Gclm和GST的mRNA表达较对照组明显降低(P<0.01),而胡黄连苷Ⅱ干预组Nrf2、NQO1、Gclc、Gclm的mRNA表达水平均较模型组明显升高(P<0.05),且呈一定程度浓度依赖性(P<0.05)。结论:PicrosideⅡ通过诱导氧化损伤L-02细胞中Nrf2表达上调,促进下游抗氧化蛋白及Ⅱ相代谢酶表达,可能是其抗氧化损伤、发挥保护肝细胞作用的机制之一。
Aim: To study the effect of Picroside Ⅱ on the mRNA expression of Phase Ⅱ metabolic enzymes in the hydrogen peroxide( H2O2 )-induced human embryo liver L-02 cells damage, and to explore the mechanism of Picroside Ⅱ protecting L-02 from oxidative stress. Method L-02 cells were treated with Picroside Ⅱ at 0,0.5,5 mmol·L-1 for 3 h as the treatment groups,and cultured with DMEM as the control and model groups,then injured by H202 (0.6 mmol·L-1) for 1h except for the control groups, and then cultured for 24h. The proliferation of L-02 cells was observed, using MTT assayThe levels of AST and ALT in cultural supernatant were determined by general methods. Real time RT-PCR were carried out to detect the mRNA expression of Nrf2, NQO1, Gcle, Gclm and GST. Results The results showed that Picroside Ⅱ could significantly inhibit the decline of cell viability induced by H2O2 ( P 〈 0.05 ). The level of AST and ALT in the model group higher than it in the control group, and the difference was statistically significant. The levels were restored remarkably in the treatment groups (P 〈 0.05). The NQO1 ,Gclc,Gclm and GST mRNA expression in the model group decreased more than these in the control group ( P 〈 0.05 ). The Nrf2, NQO1, Gclc, Gclm and GST mRNA expression in the treatment groups increased remarkably ,furthermore, these up-regulations were dose-dependent in some degree. Conclusion The results suggest that increasing the expression of the downstream peroxiredoxins and phase Ⅱ metabolic enzymes by regulating the mRNA expression of Nrf2,is the one mechanism of Picroside Ⅱ ' s preotective action on primary hepatocyte L-02 cells injure induced by H2O2.
出处
《中国中医基础医学杂志》
CAS
CSCD
北大核心
2014年第7期909-911,938,共4页
JOURNAL OF BASIC CHINESE MEDICINE
基金
国家科技重大专项资助项目-基于转化医学的艾滋病
病毒性肝炎和结核病中医证侯生物学研究(2012ZX10005001)
国家自然科学基金资助项目(81072902)
国家教育部博士点基金项目(20115132120004)