The Role Of Autophagy In Human Stellate Cell-T6 Growth And Apoptosis

Objective: To investigate the influence in the activation, proliferation, cycle and apoptosis of hepatic stellate cells and the underlying molecular mechanism after different concentrations of 3- methyladenine(3-MA) treat hepatic stellate cells in vitro.Methods: Preparate different concentrations of 3-MA and treat hepatic stellate cells. There are four groups in my study including different concentrations of 3-MA groups(3-MA 2.5mmol/l, 3-MA 5.0mmmol/l, 3-MA 10.0mmol/l) and blank control group. The cell proliferation was assessed by MTT and the cell apoptosis was detected by flow cytometry instrument in four groups of hepatic stellate cells. The expression levels of α-SMA,LC3Ⅱ,C yclin A, CyclinB1, BAX, Bcl-2 were detected by Western blot, respectively, in four groups of hepatic stel ate cel s.Results: The proliferation of cells was decreased and the cell apoptosis rate was increased after treated with 3-MA(P<0.05), and in a concentration-dependent manner, significantly higher numbers of HSCs in the phase(P<0.05);Compared with blank control group, the expression of LC3Ⅱ,α-SMA,C yclinA,C yclinB1,Bcl-2 protein in experimental groups were decreased(P<0.05),the expression of BAX protein in experimental groups were enhanced(P<0.05),and in a concentration-dependent manner.Conclusion:(1)The autophagy inhibitor 3-MA can down-regulated protein expressions of α-SMA, LC3Ⅱof hepatic stellate cells and significantly decrease the proliferation of hepatic stellate cells;(2)The autophagy inhibitor 3-MA can down-regulated the expression of C yclin A,C yclinB1 caused higher numbers in G2/M and S phase;(3)The autophagy inhibitor 3-MA can enhance the apoptosis of the cells through the pathway of inhibiting the expression of Bcl-2 and activating the BAX in vitro in a concentration-dependent manner.