| 摘要: |
| 目的探讨转化生长因子-β1（transforming growth factor-β1,TGF-β1）对实验性小鼠肝脏细胞发生上皮-间质转换（epithelial-mesenchymal transitions,EMT）的促进作用,进而加速肝纤维化（hepatic fibrosis,HF）的形成。方法建立小鼠CCl4诱导HF模型,随机分为正常对照组、HF组与TGF-β1处理组。取对数生长期肝细胞,0.25%胰蛋白酶液消化细胞,0.4%FBS/DMEM同步化处理24 h后,按TGF-β1 2.5、5、7.5 ng/ml作用24、96、144 h分组给予相应处理,相差显微镜观察细胞形态,免疫细胞组织化学及免疫荧光方法对细胞表型进行鉴定。RT-PCR及Western blot检测EMT[成纤维细胞特异性蛋白-1（fibroblast specific protein-1,FSP-1）,α-平滑肌肌动蛋白（α-smooth muscle actin,α-SMA）,E-钙黏附素（E-cadherin,E-cad）]mRNA及蛋白表达情况。结果将含有TGF-β1无血清培养基刺激原代小鼠肝细胞48 h后可见较多肝细胞死亡,细胞数量减少,形态由多边形向梭形转变,96 h后小鼠肝细胞内出现凋亡小体,活细胞数量较少且大多数呈现间质细胞梭形形态。5 ng/ml的TGF-β1作用肝细胞96 h后,小鼠肝上皮细胞标志（E-cad）mRNA及蛋白表达逐渐减低,肝间质细胞标志（FSP-1和α-SMA）mRNA及蛋白表达逐渐升高（P〈0.05）。结论 5 ng/ml的TGF-β1作用于HF的小鼠肝细胞96 h后,肝细胞发生EMT现象,TGF-β1有促进实验小鼠肝细胞的迁移、侵袭能力及细胞凋亡的作用,进而发生EMT导致HF。 |
| 关键词: 肝纤维化 转化生长因子-β1 上皮-间质转换 小鼠 |
| DOI：10. 3969/j. issnl008-0392.2014.06.003 |
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| 基金项目:国家自然科学基金（8107034） |
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| Transformation growth factor beta 1 promotes hepatocellular liver fibrosis in mice |
| BI Wan-rong1,XING Hai-lin2,JIN Cai-xia3 |
| (Dept. of Gastroenterology, The Branch of Tongji Hospital, Tongji University, Shanghai 200092, China;Dept. of General Surgery, Tongji Hospital, Tongji University, Shanghai 200065, China) |
| Abstract: |
| Objective To investigate the effect of transforming growth factor-β 1（ TGF-β 1） on experimental hepatic fibrosis（ HF） in mice. Methods Hepatic fibrosis was induced by injection of CCl4.Mice were randomly divided into normal control group,HF group and TGF-β 1 treated group. Liver cells in logarithmic growth phase were collected and digested by 0. 25% trypsin. After 24h of synchronization treatment with 0. 4% FBS / DMEM,cells were treated with different concentrations of TGF-β 1（ 2. 5,5,7. 5 ng / ml） for 24,96 h and 144 h. The cell morphology was observed by phase contrast microscope,the phenotype of cells was identified by chemical and immune fluorescence method. The mRNA and protein expression of fibroblast specific protein-1（ FSP-1）,α-smooth muscle actin（ α-SMA） and E-cadherin（ Ecad） were detected by RT-PCR and Western blotting,respectively. Results There was more liver cell death and decrease in cell numbers,after TGF-β 1 treatment for 48 h,and morphology of hepatic cells transformed from polygonal to spindle. After TGF-β 1 treatment for 96 h the cell death increased and number of living cells was further decreased and the majority cells presented as interstitial cell fusiform shape. After treatment with 5 ng / ml TGF-β 1 for 96 h the mRNA and protein expression of epithelial marker（ E-cad） in liver cells decreased,while mRNA and protein expression of hepatic interstitial cell markers（ FSP-1 and α-SMA） increased（ P〈0. 05）. Conclusion TGF-β 1 treatment can induce epithelial mesenchymal transition of liver cells,resulting in hepatic fibrosis in mice. |
| Key words: hepatic fibrosis transforming growth factor-1 epithelial mesenchymal transition mouse |
