Basic & Clinical Medicine ›› 2017, Vol. 37 ›› Issue (9): 1257-1262.

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TGF-β1-induces activation of HSC-T6 cells and epithelial-mesenchymal transition in rats

  

  • Received:2016-07-01 Revised:2016-11-03 Online:2017-09-05 Published:2017-08-28

Abstract: Objective To observe the effect of TGF-β1 on activation and epithelial mesenchymal transition in rat hepatic stellate cell-T6. Methods Adopt the MTT method to screening the optimum concentration of TGF-β1 in vitro HSC-T6 cultured. After the HSC-T6 were stimulated by TGF-β1 of 10 μg/L for 24 hours, the morphology of the cells was observed under inverted phase contrast microscope, the expression of F-actin which on behalf of cotoskeletal structure was detected by immunofluorescence staining; the expression of α-SMA and N-cadherin,vimentin,E-cadherin was measured by RT-qPCR; The changes of α-SMA,N-cadherin,vimentin and E-cadherin were assessed by Western blot after different concentrations (0 ,5 和10 μg/L) of TGF-β1 interventing HSC-T6 for 24 h. Results It had optimal cell survival rate when 10 μg/L TGF-β1 dealt with cells for 24 h. After HSC-T6 were treated with TGF-β1,cells stretched, pseudopodia increased and turn into stellate, cells connections were looser, so that represented a significantly activated state.F-actin filaments gathered to form stress and distributed along the long axis of the cells; The expression of α-SMA mRNA and vimentin mRNA in experimental group was significantly higher while E-cadherin mRNA was obviously lower than the control group(P<0.05). TGF-β1 made the protein expression of α-SMA and N-cadherin, vimentin in dose-dependent increased while E-cadherin was decreased.Conclusion TGF-β1 might induce activation and epithelial-mesenchymal transition of HSC-T6.

Key words: TGF-β1, hepatic stellate cells, activation, epithelial-mesenchymal transition

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