基于基因表达谱分析黄芪汤改善大鼠肝纤维化的分子机制研究  

Study of molecular mechanism of Huangqi Decoction improved rats liver fibrosis based on analysis of gene expression profiles

作　　者：张贵彪[1] 孙明瑜[2] 宋雅楠[1] 陆奕宇[1] 刘平[2] 苏式兵[1] 

机构地区：[1]上海中医药大学中医复杂系统研究中心,上海201203 [2]上海中医药大学附属曙光医院肝病研究所,上海201203

出　　处：《中华中医药杂志》2014年第5期1634-1639,共6页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：国家科技重大专项(No.2009ZX09311-003);国家中医药管理局重点学科项目(No.国中医药[2012]62号);上海市科委重点项目(No.12401900401);上海市教委E-研究院中医内科建设计划资助项目(No.E03008)~~

摘　　要：目的:通过观察黄芪汤对二甲基亚硝胺(DMN)诱导的大鼠肝纤维化模型的效应,分析黄芪汤对肝纤维化组织的差异基因表达谱变化,探讨黄芪汤改善肝纤维化的分子机制。方法:采用DMN建立大鼠肝纤维化模型,并以黄芪汤治疗。用药4周后处死大鼠取肝组织,观察各组大鼠肝组织病理学变化,检测羟脯氨酸(Hyp)含量,提取肝组织总RNA,用全基因组芯片检测基因表达谱变化,并进行差异表达基因、本体论(GO)、信号通路分析,实时定量RT-PCR验证mRNA表达。结果:与正常组比较,模型组肝组织病理出现显著的肝损伤和肝纤维化,肝组织Hyp含量显著升高(P<0.01)。与模型组比较,黄芪汤用药组能显著改善肝组织病理,降低肝组织Hyp含量(P<0.01)。差异基因表达谱分析显示,黄芪汤组与模型组比较后差异表达基因数量为1 011个(P<0.05,|Fold change|>1.5),其中上调431个,下调580个;富集的GO功能共有116条,主要涉及细胞外基质、细胞黏附、脂质蛋白复合物等功能;差异表达基因富集的信号通路共有70条,主要涉及细胞外基质受体作用、细胞黏附分子、黏着斑、代谢途径等。实时定量RT-PCR验证的基因表达与基因芯片检测结果趋势一致。结论:黄芪汤通过多基因、多功能、多信号通路的调节,改善了DMN诱导的大鼠肝纤维化病变。Objective: To investigate the molecular mechanism of HuangqfiDecoction(HQD) improved the liver fibrosis induced by dimethylnitrosamine(DMN) in rats, through the analysis of differential gene expression profiles of liver fibrosis tissues. Methods: Liver fibrosis in rats was induced by intraperitoneally injection of DMN, and treated by HQD for 4 weeks. Then the pathological changes of hepatic tissue were observed, hydroxyproline(Hyp) content were measured, and total RNA of liver tissue were extracted. The gene expression profile was detected by whole genome chip testing, then differentially expressed genes(DEGs), gene ontology(GO) and signal pathways were analyzed. The mRNA expressions were verified by real-time RT-PCR. Results: Compared with the normal group, the liver tissue in model group presented the pathological changes of liver damage and liver fibrosis, and the Hyp content of liver tissue significantly increased(P<0.01). Compared with the model group, the HQDtreated group markedly improved the pathological changes of liver tissue, significantly reduced the Hyp content(P<0.01). The DEGs between HQD-treated and model control(P<0.05, |Fold change|>1.5) were 1 011, included 431 up-regulated and 580 downregulated genes. The GO enrichment analysis(P<0.05) showed 116 functions, included extracellular matrix, cell adhesion, proteinlipid complex, and so on. The signal pathway analysis showed 70 enriched pathways(P<0.05), included ECM-receptor interaction, cell adhesion molecules, focal adhesion, metabolic pathways, and so on. The verification of mRNA expression by Real-time RTPCR was consistent with that of microarray. Conclusion: HQD improves the pathological changes of liver fibrosis through the regulation of multiple genes and multiple signal pathways in rats.

关 键 词：黄芪汤 肝纤维化 基因表达谱 分子机制 

分 类 号：R285.5[医药卫生—中药学]