大鼠CCl_4所致肝纤维化自发逆转的基因表达谱研究  

Gene expression profile analysis of spontaneous reversion of CCl_4-induced hepatic fibrosis in rats

作　　者：潘勤[1] 谢渭芬[1] 张忠兵[1] 张新 韩泽广 

机构地区：[1]中国人民解放军第二军医大学附属长征医院消化内科,上海市200003 [2]国家人类基因组南方研究中心,上海市201203

出　　处：《世界华人消化杂志》2005年第13期1544-1549,共6页World Chinese Journal of Digestology

基　　金：中国博士后科学基金资助项目;No.2004036325~~

摘　　要：目的:利用cDNA微阵列杂交筛选肝纤维化自发逆转过程中的差异表达基因,从而揭示纤维化逆转的基因表达谱. 方法:通过四氯化碳(carbon tetrachloride,CCl4)注射SD大鼠8 wk,随后停药6 wk建立肝纤维化自发逆转的动物模型.提取纤维化逆转过程中不同时间段的肝组织mRNA,与cDNA微阵列杂交,筛选表达水平明显上调或下调的差异基因,并结合Cluster分析等进行分类.此外选择α共核蛋白、A-raf、早老素2及β肌动蛋白作RT-PCR以验证检测结果. 结果:CCl4诱导的肝纤维化在停止毒剂注射后出现进行性消退.cDNA微阵列杂交证实,肝纤维化消退过程中共有254条基因出现转录水平改变,占基因总数的21.59%. 其中8,10,12,14 wk的差异表达基因分别为54, 85,97,132条.筛选所得的肝纤维化逆转相关基因主要涉及代谢相关酶、脂肪代谢相关蛋白、离子通道、转录因子、胃肠激素及受体、MAPK及PI3k/Akt信号转导通路等.RT-PCR检测与cDNA微阵列杂交一致,证实了结果的可靠性. 结论:肝纤维化逆转过程中基因表达谱发生显著改变, 主要涉及代谢相关酶、转运蛋白/协同转运蛋白、胃肠激素及受体、脂蛋白/脂肪酸结合蛋白、转录因子/ 核因子、MAPK信号转导通路等6类基因.AIM: To screen the differentially expressed gene in the spontaneous reversion of hepatic fibrosis induced by carbon tetrachloride (CCI4), and to reveal the gene expression profile in this process. METHODS: Animal model of hepatic fibrosis with spontaneous reversion was created in SD rats by injection of carbon tetrachloride (CCI4) for 8 weeks and then withdrawing for 6 weeks. The mRNA of liver tissues was extracted at different time spots in the process of reversion. Then cDNA microarray was used to screen the differentially expressed genes. Finally the products were subjected to hierarchical clustering and confirmed by semi-quantitative reverse transcription polymerase chain reaction (RT-PCR) of α-synuclein, A-raf, presenilin-2 and β-actin. RESULTS: Hepatic fibrosis was progressively reversed after stopping injection of CCI4. All together, there were 254 (21.59%) genes that changed at the transcription level. Meanwhile, 54, 85, 97 and 132 genes were identified differentially expressed in the 8th, 10th, 12th and 14th week, respectively. After confirmed by RT-PCR, the differentially expressed were associated with metabolic enzymes, ion channels, transcription factors, gastrointestinal hormones and their receptors, mitogen-activated protein kinase (MAPK) and PI3k/Akt signaling pathway. CONCLUSION: The gene expression profile is significantly changed in the spontaneous recovery of CCl4-induced hepatic fibrosis. Genes closely related to the spontaneous recovery are associated with metabolic enzymes, transporter/symporter proteins, gastrointestinal hormones/ receptors, lipoprotein/fatty acid binding proteins, transcription factor/nuclear factors, and MAPK signal pathway.

关 键 词：基因表达谱 CCL4 cDNA微阵列杂交 自发 MAPK信号转导通路 RT-PCR检测 肝纤维化逆转 脂肪酸结合蛋白 差异表达基因 转录因子 胃肠激素 转运蛋白 β肌动蛋白 杂交筛选 四氯化碳 SD大鼠 mRNA 动物模型 差异基因 检测结果 早老素2 

分 类 号：R575.2[医药卫生—消化系统]