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目的研究14-3-3蛋白在中枢神经系统小神经胶质细胞中的表达分布情况及LPS刺激后其表达变化;探讨14-3-3蛋白与小神经胶质细胞释放炎症因子的关系。方法应用小鼠小神经胶质细胞株BV-2作为体外小神经胶质细胞的模型,以LPS刺激其活化,应用免疫荧光检测14-3-3蛋白ζεθγ亚型在BV-2细胞中的表达、免疫印迹技术检测14-3-3蛋白在LPS刺激后的表达变化,ELISA检测IL-1β的表达。结果14-3-3ζεθγ蛋白亚型在BV-2细胞中均有表达,主要分布在细胞浆;LPS刺激BV-2细胞活化后释放IL-1β,同时14-3-3蛋白表达下降。结论BV-2细胞高表达14-3-3蛋白,LPS刺激后表达下降,提示14-3-3蛋白参与了中枢神经系统免疫应答的调节。
Objective To study the expression and distribution of 14-3-3 protein in the central nervous system microglia and its expression after LPS stimulation and to explore the relationship between 14-3-3 protein and the release of inflammatory cytokines by microglia. Methods Mouse microglia cell line BV-2 was used as a model of microglia in vitro and stimulated by LPS. The expression of ζεθγ subunit of 14-3-3 protein in BV-2 cells was detected by immunofluorescence Western blotting was used to detect the expression of 14-3-3 protein after LPS stimulation. The expression of IL-1β was detected by ELISA. Results The 14-3-3 ζεγ protein subtype was expressed in BV-2 cells and mainly distributed in the cytoplasm. After LPS stimulated the activation of BV-2 cells, IL-1β was released and the expression of 14-3-3 protein decreased. Conclusions The expression of 14-3-3 protein in BV-2 cells is high, which is down-regulated after stimulation with LPS, suggesting that 14-3-3 protein is involved in the regulation of central nervous system immune response.