摘要
目的探讨ZEB1在大鼠肝纤维化模型和肝星状细胞中的动态变化和意义。方法健康雄性SD大鼠30只,分为健康对照组和模型组,模型组按造模后不同时间点分为2、4、6、8周4个亚组,每组6只。模型组腹腔注射二甲基亚硝胺(DMN)诱导形成大鼠肝纤维化模型,测定不同时间点ALT、AST的变化,肝组织经HE、Masson染色,光学显微镜下观察;采用RT-PCR检测不同时间点肝脏ZEB-1mRNA的表达。大鼠HSCT-6细胞用不同浓度TGF-β1因子刺激48h后提取RNA,检测不同TGF-β1浓度下ZEB-1mRNA的表达。各组间均数比较采用单因素分析。结果 DMN腹腔注射成功诱导大鼠肝纤维化,4、6周纤维化明显。模型组血清ALT、AST在2周开始升高,4周达到最高峰,并明显高于健康对照组。ZEB-1mRNA在正常肝组织中少量表达,模型组表达逐渐升高,4周表达最高,随后又逐渐降低,相关性分析显示与TGF-β1呈正相关。HSC-T6细胞中,不同浓度TGF-β1刺激后,ZEB-1mRNA呈浓度梯度变化,随着TGF-β1浓度的升高,ZEB-1表达量增加。结论 ZEB-1可能参与肝纤维化的发生,并与肝纤维化的发展密切相关。
Objective To explore the dynamic expression of ZEB - 1 in rat model of hepatic fibrosis and hepatic stellate cells induced by TGF - β1. Methods A total of 30 healthy male SD rats were randomly divided into normal control group and model group,and the model group was further divided into four subgroups according to different time points： subgroups of 2,4,6 and 8weeks with six rats iu each group. The rat model of hepatic fibrosis was induced by dimethylnitrosamine（DMN）. The serum of alanine aminotransferase （ ALT）, aspertatr aminotrabsferase（AST） were detected dynamically after 2,4,6,8weeks of infection. The liver tissues were observed under optical microscope after HE and Masson staining. ZEB - 1 mRNA was detected by reverse transcriptase polymerase chain reaction （ RT - PCR）. Rat HSC T -6 cell was stimulated with different concentrations of TGF -β lfactor, then the RNA was extracted to detect the expression of ZEB - 1 mRNA of different concentration of stimulus. Results The hepatic fibrosis model was successfully induced by DMN infection and the level of hepatic fibrosis were the most obvious at 4 and 6weeks. The serum levels of ALT and AST were all increased after 2 week of infection and peaked at week 4 which were all significantly higher than those in control group. The expression of ZEB - 1 mRNA was signif- icantly increased than that in control group. And the expression of ZEB - 1 was closely correlated with the TGF - β1 expression. In HSC - T6 cells, the ZEB - lmRNA gradually increased follow with the increased TGF - β1 concentration. Conclusion The ZEB - 1 may participate in the development of hepatic fibrosis ,which is closely correlated with the progression and severity of liver fibrosis.
出处
《医学研究杂志》
2013年第1期118-121,共4页
Journal of Medical Research
