乌索酸体外通过激活Nrf2信号通路抑制人肝星状细胞的增殖和细胞外基质相关基因的表达  

Ursolic Acid Inhibits the Proliferation of Human Hepatic Stellate Cells and the Expression of Extracellular Matrix-Related Genes by Activating Nrf2 Signaling Pathway in Vitro

作　　者：白庆云 Bai Qingyun(School of Chemical and Biomedical Engineering,Yichun University,Jiangxi Key Laboratory of Natural Drug Active Ingredients,Yichun 336000,China)

机构地区：[1]宜春学院化学与生物工程学院江西省天然药物活性成分研究重点实验室,宜春336000

出　　处：《世界科学技术-中医药现代化》2020年第1期140-146,共7页Modernization of Traditional Chinese Medicine and Materia Medica-World Science and Technology

基　　金：江西省科技厅自然科学基金面上项目(20132BAB205087):乌索酸对人HSC表型转化、ECM以及MMPs/TIMPs系统的作用,负责人,白庆云;江西省教育厅科技项目一般项目(GJJ170905):自噬底物p62蛋白在绿原酸拮抗对乙酰氨基酚肝损伤中的作用,负责人:白庆云.

摘　　要：目的乌索酸(Ursolic acid,UA)具有抗肝纤维化作用,但是其机制尚不完全清楚。由于肝纤维化与氧化应激有关,因此本文通过对乌索酸抗氧化作用的研究以阐释其抗肝纤维化机制。方法实验用实时定量聚合酶链反应(real time polymerase chain reaction,RT-PCR)法和Western blot法检测氧化应激状态下LX2中α平滑肌肌动蛋白(alpha muscle actin,α-SMA)、Ⅰ型胶原(collagen type I,COL1A1)、Ⅲ型胶原(collagen type Ⅲ,COL3A1)、基质金属蛋白酶(matrix metalloproteinase 2,MMP2)和基质金属蛋白酶组织抑制因子(tissue inhibitor of metalloproteinase 1,TIMP1)mRNA和蛋白表达的变化以及UA对这些变化的影响;通过测定还原型谷胱甘肽(glutathione,GSH)和丙二醛(malondialdehyde,MDA)的含量以观察UA的抗氧化作用,并检测了核因子E2相关因子2(NF-E2-related factor 2,Nrf2)及其下游基因血红素加氧酶(heme oxygenase 1,HO1)蛋白表达的变化,探察UA抗肝纤维化作用的可能机制。结果实验结果显示H2O2可引起LX2的活化和氧化应激,使细胞中的ECM mRNA表达增加。UA可抑制LX2的活化,减少细胞外基质(extracellular matrix,ECM)的表达,显著升高GSH含量,降低MDA含量,并增加Nrf2的转核和HO1的表达。结论实验结果提示UA通过激活Nrf2信号通路发挥抗氧化作用从而抑制LX2的活化和ECM的产生。Objective Ursolic acid has anti-hepatic fibrosis effect,but its mechanism is not completely clear.Because liver fibrosis is related to oxidative stress,so this paper aims to elucidate the anti-fibrosis mechanism of ursolic acid through the study of its antioxidant effect.Methods We detected the mRNA and proteins expression of alpha muscle actin(alpha-SMA),collagen type I(COL1A1),collagen type III(COL3A1),matrix metalloproteinase(MMP2)and tissue inhibitor of metalloproteinase 1(TIMP1)in LX2 under oxidative stress by RT-PCR and Western blot.At the same time,we observed the effect of UA on these proteins and genes.Furthermore,glutathione(GSH)and malondialdehyde(MDA)were measured to observe the antioxidant activity of UA.We also detected the protein expression of Nrf2 and its downstream gene heme oxygenase 1(HO1)to explore the possible anti-fibrosis mechanism of UA.Results The results showed that H2O2 induced LX2 activation and oxidative stress,increased the production of ECM.UA could inhibit the activation of LX2 and reduce the production of ECM.The same while,UA significantly increased the content of GSH and reduced the content of MDA.Furthermore,UA increased the expression of nuclear Nrf2 and HO1.Conclusion Our study suggested that UA exerted antioxidation by activating Nrf2 signaling pathway to inhibit LX2 activation and ECM production.

关 键 词：乌索酸 肝星状细胞LX2 肝纤维化 H2O2 抗氧化 NRF2 

分 类 号：R2-031[医药卫生—中西医结合]