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目的:探讨肿瘤坏死因子-α刺激的星形胶质细胞对神经元的作用。材料和方法:体外纯化培养大鼠海马星形胶质细胞,采用反相高效液相法测定TNFα刺激后细胞培养液内谷氨酸的含量;将TNFα刺激后的星形胶质细胞条件培养液(Astrocytic Conditioned Medium,ACM)作用于培养的海马神经元,运用免疫细胞化学方法和图像分析技术研究神经元NF-κBp65和谷氨酸的表达。结果:(1)TNFα可明显促进星形胶质细胞释放谷氨酸;(2)ACM作用15min即可诱导神经元NF-κBp65的核表达,30min达高峰,180min恢复至对照水平;(3)ACM作用60min可使谷氨酸免疫反应阳性神经元平均光密度明显升高,持续至240min。结论:TNFα刺激的星形胶质细胞可通过释放谷氨酸等可溶性物质使神经元快速激活、兴奋性升高。
Objective: To investigate the effect of tumor necrosis factor-α-stimulated astrocytes on neurons. MATERIALS AND METHODS: Hippocampal astrocytes were purified from rat hippocampus in vitro. The content of glutamate in the culture medium was measured by reverse-phase high-performance liquid chromatography (RP-HPLC). The astrocyte-conditioned medium (Astrocytic Conditioned Medium, ACM) on cultured hippocampal neurons, using immunocytochemistry and image analysis techniques to study the expression of NF-κBp65 and glutamate in neurons. (2) The expression of NF-κBp65 in neurons could be induced by ACM for 15min, reaching the peak at 30min and returned to the control level after 180min; (3) ACM effect of 60min can significantly increase the average optical density of glutamate immunoreactive neurons, until 240min. CONCLUSION: TNFα-stimulated astrocytes can rapidly activate neurons by releasing soluble substances such as glutamate, resulting in increased excitability.