目的:观察大鼠肝纤维化形成过程中成纤维细胞激活蛋白(fibroblast activation protein, FAP)的动态表达变化特点。方法:Wistar雄性大鼠分为正常对照组和模型组。模型组ip 0.5%二甲基亚硝胺复制肝纤维化模型, 于造模1、2、3周分别收集肝组织标本, 造模4周后处死大鼠, 收集血清和肝组织标本。全自动生化仪测定丙氨酸氨基转移酶(ALT)、天门冬氨酸氨基转移酶(AST)、总胆红素(TBIL)、谷氨酰转肽酶(GGT)和白蛋白(ALB), HE、天狼猩红染色观察病理形态, 试剂盒测定肝组织羟脯氨酸;real-time PCR和western blotting法检测肝组织FAP基因和蛋白表达。结果:模型组肝功能较正常组明显下降, 组织病理形态学和羟脯氨酸测定显示模型组肝损伤明显, 纤维化形成。real-time PCR和western blotting显示FAP基因和蛋白随造模时间延长表达逐渐增高。结论:FAP伴随大鼠肝纤维化模型形成逐渐增加, 与肝纤维化形成密切相关。
Objective: To observe the dynamic changes in fibroblast activation protein (FAP) level during the formation of liver fibrosis induced by dimethylnitrosamine in rats. Methods: Twenty-four male Wistar rats were randomly divided into normal control group and model group. Rats in the model group were intraperitioneally injected with 0.5% dimethylnitrosamine for 4 weeks. During fibrosis processing, liver sample was collected from selected rats at 1, 2 and 3 weeks. Other rats were sacrificed after collecting serum and liver sample. Liver function, including ALT, AST, GGT, TBiL, and Alb in serum was measured using the biochemical analyzer. Meanwhile, liver tissues were stained with HE and Sirius red and observed under a light microscope for indentifying the pathological changes. Fap expression in both protein and RNA levels was detected using Western blotting or real-time PCR method respectively. Results: Rats in the model group were underwent impaired liver functions, which were showed in evaluated ALT, AST, GGT and TBiL levels and declined Alb content. In comparison with the normal control group, liver tissue of the model group showed typical pathological changes of fibrosis as well as an obviously increased hydroxyproline content. FAP protein and RNA expression was increased, with a time-dependent manner. Conclusion: FAP expression level is closely related with liver fibrosis degree, which is confirmed by the result of progressive rising FAP expression level in the fibrosis process.