基于基因表达谱分析扶正化瘀方改善大鼠肝纤维化的分子机制  

Study on molecular mechanism of improving rat liver fibrosis with Fuzheng Huayu Formula based on gene profile

作　　者：董姝[1] 陈启龙[1] 冯琴[2] 胡义扬[2] 刘平[3] 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,上海201203 [3]上海中医药大学E研究院,上海201203

出　　处：《中华中医药杂志》2015年第5期1812-1817,共6页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：国家科技重大专项(No.2009ZX09311-003);上海市科委重点项目(No.12401900401);上海市教委E研究院建设项目(No.E03008)~~

摘　　要：目的:通过观察扶正化瘀方对四氯化碳(CCl4)诱导的大鼠肝纤维化模型的效应,分析扶正化瘀方对肝纤维化组织的差异基因表达谱变化,探讨扶正化瘀方改善肝纤维化的分子机制。方法:采用CCl4建立大鼠肝纤维化模型,以扶正化瘀方进行治疗。用药后3周处死大鼠,取肝组织,进行HE染色和天狼星红染色,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,用全基因芯片检测基因表达谱,据此进行差异基因筛选,本体论(GO)、信号通路分析。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)。与模型组相比,扶正化瘀方组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:扶正化瘀方组与模型组相比,差异表达基因数量为397个(P<0.05,|Fold Change|>1.5),其中上调的有195个,表达下调的有202个;富集的GO功能共有160条,涉及对成纤维细胞增殖的正向调节、对上皮细胞增殖的调节、对细胞外基质刺激物的应答、维生素A/视黄醇代谢过程等。参与调节的信号通路有17条,包括药物代谢、细胞色素P450外源性物质的代谢、花生四烯酸代谢等。结论:扶正化瘀方主要从调控纤维母细胞、上皮细胞增殖等信号通路改善CCl4诱导的大鼠肝纤维化病变。Objective： To analyze the difference in gene expression profile changes of liver fibrosis treated with Fuzheng Huayu Formula and investigate the molecular mechanism of improving liver fibrosis with Fuzheng Huayu Formula through observing the effect of treating rat liver fibrosis induced by CC14 with Fuzheng Huayu Formula. Methods： Rat liver fibrosis models were established by using injection of CC14, and rat liver fibrosis models were treated with Fuzheng Huayu Formula. After treatment for 3 weeks, rats were sacrificed, and the pathologic changes of liver tissues were observed by using HE staining and Sirius red staining. The contents of HyP were detected. Gene expression profile was detected by using gene chip, now therefore, screened the differentially expressed genes and analyzed ontology （GO） and signal path. Results： Compared with control group, liver tissues of rats in model group appeared significant damage and fibrosis, and HyP contents in liver were increased significantly （P〈0.01）. Compared with model group, Fuzheng Huayu Formula could improve liver pathological changes and reduce the HyP contents in liver tissue （P〈0.01）. Analysis of difference in gene expression profile showed that there were 397 differential expression genes in Fuzheng Huayu group compared with model group （P〈0.05, ｜Fold Change｜〉1.5）, and 195 geneswere up-regulated and 202 genes were down-regulated. There were 160 enrichment GO functions, and the GO functions involved regulation of fibroblast proliferation and epithelial cell, reply of extracelluar matrix stimulation and regulation of metabolism process of Vitamin A/retionl. There were 17 signal paths involved in the regulation, which included drug metabolism, xenobiotics metabolism of P450, arachidonic acid metabolism and so on. Conclusion： Fuzheng Huayu Formula could improve liver fibrosis process of rats induced by CCI through regulation of fibroblast and signal pathway of epithelial cell proliferation.

关 键 词：扶正化瘀 肝纤维化 基因表达谱 羟脯氨酸 分子机制 

分 类 号：R285.5[医药卫生—中药学]