Effect of lipopolysaccharide on IL-6/STAT3 signal pathway in human intrahepatic biliary epithelial cells / 安徽医科大学学报

ABSTRACT

Objective To investigate the expression of IL-6/STAT3 signal pathway induced by lipopolysaccharide (LPS) on human intrahepatic biliary epithelial cells (HiBECs) in vitro.Methods The HiBECs were cultured in vitro.The cells were stimulated by LPS at different concentrations (0,0.1,1,2,4,8 μg/ml) and different time (24,48,72 h).The expression of IL-6 was measured by ELISA to determine the best concentration and time of LPS.Then we chose the best concentration and time of LPS to intervene the cells in the further experiments.The levels of mRNA expression of c-Myc and Mcl-1 were detected by fluorescence quantitative reverse transcrption PCR (RT-qRCR).The proportion of p-STAT3/STAT3 protein and the expression of c-Myc and Mcl-1 protein in HiBECs were analyzed by Western blot.Results LPS upregulated the expression of IL-6, and reached its peak at 4 μg/ml and 24 h(F=16.492,17.763,P<0.001,P<0.01).LPS increased the mRNA expression of c-Myc and Mcl-1(P<0.01).Meanwhile, The ratio of p-STAT3/STAT3 (P<0.05) protein and the expression of c-Myc (P<0.001) and Mcl-1 (P<0.05) protein were upregulated after stimulating with LPS.Conclusion The IL-6/STAT3 signal pathway can be activated by LPS in HiBECs, and LPS may promote the expression of c-Myc and Mcl-1 through this signal pathway.