解放军医学杂志2017，Vol.42Issue(5)：383-388,6.DOI:10.11855/j.issn.0577-7402.2017.05.05
熊果酸对TGF-β1诱导肝细胞凋亡的抑制作用及其机制
Inhibiting effect of ursolic acid on hepatocyte apoptosis induced by TGF-β1 and its mechanism
摘要
Abstract
Objective To study the effect of ursolic acid (UA) intervention on hepatocyte apoptosis induced by TGF-β1 and its potential mechanism.Methods Primary hepatocytes were extracted from healthy SD rats by in situ perfusion,cultured for 12-24h,then randomly divided into the following groups:blank control group,UA control group (UA 25μmol/L),TGF-β1 group (TGF-β1 2.5ng/ml),UA intervention group (UA 25μmol/L and TGF-β1 2.5ng/ml),DPI intervention group (DPI 0.5μmol/L and TGF-β1 2.5ng/ml).Each group was treated with drugs for corresponding time and their proliferation and apoptosis were detected by flow cytometry,the expression of CD95 (Fas) mRNA was analyzed by RT-qPCR,the expression of protein CD95 and membrane translocation of NADPH oxidase (NOX) subunit p47Phox were analyzed by Western blotting,and the reactive oxygen species (ROS) generation in primary hepatocytes was analyzed with reactive oxygen detection kit.Results UA intervention at 30min before TGF-β1 stimulating hepatocytes markedly reduced hepatocyte apoptosis (63.97 ± 3.19 vs 80.53 ± 1.56,P＜0.01) and promoted hepatocyte proliferation (18.67 ± 1.60 vs 10.83 ± 2.03,P＜0.01).UA intervention notably down-regulated the expressions of CD95 mRNA and protein (1.28 ± 0.15 vs 2.40 ± 0.25,P＜0.01;1.05 ± 0.15 vs 1.37 ± 0.18,P＜0.05),restrained membrane translocation of p47phox (1.13 ± 0.12 vs 1.76 ± 0.22,P＜0.01),and decreased ROS level in primary hepatocytes induced by TGF-β1 (2.12 ± 0.45 vs 3.23 ± 0.53,P＜0.01).Conclusion The mechanism of UA inhibiting hepatocyte apoptosis induced by TGF-β1 is likely to be that UA intervention reduced hepatocyte apoptosis by inhibiting NOX activation and decrease generation of ROS so as to down-regulate expression of CD95 in hepatocytes.关键词
熊果酸/肝细胞/转化生长因子β1/NADPH氧化酶/细胞凋亡Key words
ursolic acid/hepatocytes/transforming growth factor β1/NADPH oxidase/apoptosis分类
医药卫生引用本文复制引用
周娟娟,何文华,甘达凯,张望,彭阿平,汪安江,李弼民,朱萱..熊果酸对TGF-β1诱导肝细胞凋亡的抑制作用及其机制[J].解放军医学杂志,2017,42(5):383-388,6.基金项目
国家自然科学基金(81160061,81260082)This work was supported by the National Natural Science Foundation of China (81160061,81260082) （81160061,81260082）
