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摘要
Objective: This experiment starts with the role of Epithelial-mesenchymal transition(EMT)in BA extrahepatic bile duct fibrosis and its regulatory mechanism, to investigate the induction of TGF-β1 on extrahepatic bile duct EMT and the mechanism of HGF in improving extrahepatic bile duct fibrosis by regulating EMT. Methods: Extrahepatic bile duct was isolated from BALb/c mice and extraction of extrahepatic bile duct epithelial cells was performed. Immunofluorescence was used to identified the surface markers of extrahepatic bile duct epithelial cells. The proliferation of extrahepatic bile duct epithelial cells was observed after culture.P3 generation extrahepatic bile duct epithelial cells were used for experimental grouping, normal control group(control group), induced EMT group(TGF-β1 group), HGF intervention group (HGF+ TGF-β1 group), HGF control group (HGF group) and HGF inhibition group (TGF-β1+ HGF+ SU11274 group) for a total of 5 groups, with the same incubation time of each factor for 48h. Finally the obtained cells were used to determine the expression of fibrosis related proteins and genes (CK19, E-cad, α-SMA, S100A4) by western blotting. The data were analyzed by Statistics. Results: Western blot experiment showed that: Compared with the control group, TGF-β1 could induce down-regulation of epithelial cell markers CK19 (0.75±0.05) and E-cad (0.66±0.06) in extrahepatic bile duct epithelial cells, while up-regulation of mesenchymal cell markers α-SMA (1.47±0.05) and S100A4 (1.49±0.06), the differences between the groups were statistically significant (P<0.05); Compared with TGF-β1 group, the expressions of CK19 (1.20±0.05) and E-cad (1.12±0.06)were up-regulated and α-SMA(0.71±0.04) and S100A4 (0.82±0.07) were down-regulated after HGF intervention, the differences between the groups were statistically significant (P<0.05); Compared with HGF intervention group, after HGF inhibitor SU11274 was applied, CK19 (0.81±0.05) and E-cad (0.99±0.06) expressions were down-regulated, α-SMA (1.25±0.05) and S100A4 (1.03±0.07) expressions were up-regulated, with the exception of E-cad, and the differences between the groups were statistically(P<0.05). Conclusions: TGF-β1 can induce epithelial-mesenchymal transformation of extrahepatic bile duct epithelial cells. HGF can improve extrahepatic bile duct fibrosis by regulating TGF-β1 induced EMT.
