The Effects Of Sept4in LX-2Cells Stimulated By LPS

Objective To observe the expression of Sept4in human LX-2cells stimulated by LPS and investigate its function in the process of HSC activation.Methods1. The human LX-2cells were stimulated with LPS and the cells were harvested to detect PCNA, a-SMA and active caspase-3proteins by Western blot.2. Western blot and RT-PCR were used to detect the changes of Sept4mRNA and protein expression in LX-2cells stimulated by LPS.3. The HTA125antibody was used to analyze reasons for the changes of Sept4, a-SMA and Smad4protein expression. Cultured cells pre-transfected with the Sept4siRNA were stimulated by LPS in order to observe the influence of Sept4on HSC activation.Results1. In LX-2cells, LPS could induce the alteration of a-SMA, PCNA and active caspase-3protein expression.2. The expression of Sept4in the activated LX-2cells was regulated by LPS. 3. LPS regulated Sept4expression in LX-2cells via TLR4. The HTA125antibody could also regulated a-SMA and Smad4expression in LX-2cells treated by LPS. After transfecting Sept4siRNA into LX-2, we found that a-SMA and Smad4protein expression was regulated through downregulation of Sept4protein level in LX-2cells stimulated by LPS.Conclusions1. The expression of Sept4in the activated LX-2cells was regulated by LPS via TLR4.2. Sept4may be involved in the TLR4-TGF-(3-smad4process of liver fibrosis and activation of HSC by LPS.