EFFECTS OF LIPOPOLYSACCHARIDE ON THE EXPRESSION OF SRC-SUPPRESSED C KINASE SUBSTRATE IN CULTURED ASTROCYTES / 解剖学报

ABSTRACT

Objective To investigate the effects of lipopolysaccharide(LPS) on Src-suppressed C Kinase Substrate(SSeCKS) in cultured astrocytes. Methods Purified astrocytes were randomly divided into three groupscontrol group,singly stressed and multiple stressed groups.The expression of SSeCKS was detected by Real time RT-PCR and Western blotting,while immunocytochemistry was used to investigate the subcellular localization of it. Results Real time RT-PCR indicated that,after 12 hours incubation,both 100 ?g/L and 1mg/L LPS were able to elevate the levels of SSeCKS mRNA compared with control group,while 1mg/L LPS did not have a stronger effect than 100 ?g/L.Western blotting analysis showed 100 ?g/L LPS significantly increased the expression of SSeCKS.In time response experiments,the levels of SSeCKS expression enhanced three hours after the stimulation,peaked at the sixth hour,coincident with its rapid phosphorylation,and remained high until the 24th hour.Immunocytochemistry suggested a perinuclear translocation of SSeCKS,while the PKC inhibitor RO-31-8220 blocked it.Conclusion In cultured astrocytes,LPS can enhance the expression of SSeCKS,increase its phosphorylation level and change its subcellular localization.These effects are correlated with the PKC pathway,which indicates SSeCKS might participate in the signal transduction of inflammation in cultured astrocytes.