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[摘要]
目的:觀察大鼠肝纖維化形成過程中成纖維細胞激活蛋白(fibroblast activation protein, FAP)的動態(tài)表達變化特點。方法:Wistar雄性大鼠分為正常對照組和模型組。模型組ip 0.5%二甲基亞硝胺復制肝纖維化模型, 于造模1、2、3周分別收集肝組織標本, 造模4周后處死大鼠, 收集血清和肝組織標本。全自動生化儀測定丙氨酸氨基轉移酶(ALT)、天門冬氨酸氨基轉移酶(AST)、總膽紅素(TBIL)、谷氨酰轉肽酶(GGT)和白蛋白(ALB), HE、天狼猩紅染色觀察病理形態(tài), 試劑盒測定肝組織羥脯氨酸;real-time PCR和western blotting法檢測肝組織FAP基因和蛋白表達。結果:模型組肝功能較正常組明顯下降, 組織病理形態(tài)學和羥脯氨酸測定顯示模型組肝損傷明顯, 纖維化形成。real-time PCR和western blotting顯示FAP基因和蛋白隨造模時間延長表達逐漸增高。結論:FAP伴隨大鼠肝纖維化模型形成逐漸增加, 與肝纖維化形成密切相關。
[Key word]
[Abstract]
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.
[中圖分類號]
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