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目的研究炎性介质脂多糖(LPS)、γ干扰素(IFN-γ)刺激离体星形胶质细胞的效应。方法LPS、IFN-γ刺激离体培养纯化的小鼠星形胶质细胞,ELISA检测细胞分泌TNF-α,Griess法测定NO浓度,激光共聚焦扫描显微镜(CLSM)检测细胞内游离钙浓度([Ca2+]i)。结果LPS(500ng/ml)、IFN-γ(100U/ml)刺激星形胶质细胞24h后,NO和TNF-α的分泌显著升高,联合应用有协同作用;延长IFN-γ作用时间,分泌量升高。CLSM检测LPS、IFN-γ刺激24h后的星形胶质细胞[Ca2+]i的荧光值,显示荧光相对值比正常细胞明显升高;LPS、IFN-γ急性刺激正常培养细胞发现[Ca2+]i振荡上升;而作用于IFN-γ刺激24h后的细胞,[Ca2+]i上升明显减缓。结论LPS、IFN-γ可以活化星形胶质细胞,表现为[Ca2+]i升高,TNF-α和NO分泌上调。
Objective To investigate the effects of inflammatory mediators lipopolysaccharide (LPS) and interferon-γ (IFN-γ) on astrocyte in vitro. Methods Purified mouse astrocytes were stimulated by LPS and IFN-γ, TNF-α was secreted by ELISA, NO concentration was measured by Griess method, and intracellular free calcium concentration was detected by confocal laser scanning microscopy (CLSM) Ca2 +] i). Results After stimulated with LPS (500ng / ml) and IFN-γ (100U / ml) for 24 h, the secretion of NO and TNF-α were significantly increased, and synergistic effect was observed. The effect of prolonged IFN- Increase in volume. The fluorescence value of [Ca2 +] i in astrocytes stimulated by LPS and IFN-γ for 24 hours showed that the relative fluorescence intensity of LPS and IFN-γ was significantly higher than that of normal cells. LPS and IFN-γ acutely stimulated [Ca2 +] i Oscillation increased; while the cells treated with IFN-γ 24h, the [Ca2 +] i increased significantly slowed down. Conclusion LPS and IFN-γ can activate astrocytes, showing the increase of [Ca2 +] i, up-regulation of TNF-α and NO secretion.