关键词: 细胞脂多糖, Src抑制的蛋白激酶C的底物, 星形胶质细胞, 蛋白激酶C, 免疫印迹, 免疫组织化学, 大鼠

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 groups:control 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 μg/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 μg/L. Western blotting analysis showed 100 μg/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 BR>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 inflammatio

Key words: LPS, SSeCKS, Astrocytes, PKC, Western blotting, Immunohistocytochemistry, Rat