去铁敏对大鼠星型胶质细胞谷氨酸毒性的保护作用

Protective effect of deferroxamine on glutamate induced neurotoxicity in cultured astrocytes

目的:观察去铁敏对谷氨酸所致星型胶质细胞损伤的保护作用。方法:建立原代培养的星型胶质细胞谷氨酸毒性模型,采用免疫组化、Hoechst染色检测细胞形态及核固缩情况。CCK-8法检测细胞活力。生化法检测羟自由基、MDA变化。用显微荧光测量技术监测神经元内钙信号的动态变化。结果:随着谷氨酸浓度的增加细胞损伤也逐渐加重。与对照组相比,去铁敏处理后细胞对0.5,1.0,5.0mmol/L谷氨酸损伤形态保持良好;细胞核固缩率减少,分别为(10.44±5.03)%,(7.93±5.04)%和(11.01±3.73)%(P<0.05);细胞活力下降减轻,分别为(94.72±2.20)%(P<0.05),(82.49±1.94)%和(67.94±5.55)%(P<0.01)。去铁敏处理后羟自由基水平下降,为10.17±1.79(P<0.01)。MDA水平下降,为7.36±1.47(P<0.01)。去铁敏预处理后钙离子增高的幅度及出现钙离子变化的细胞数较对照有明显减少。结论:去铁敏能减轻谷氨酸导致的星型胶质细胞损伤,能减少谷氨酸导致的星型胶质细胞内钙浓度的升高及自由基水平,这可能是去铁敏保护作用的机理之一。

Objective：To investigate the protective effect of deferroxamine on glutamate-induced injury in cultured astrocytes.Methods：Primarily cultured astrocytes injury were induced by glutamate and then observed the protective effect of defferroxamine.The morphological change was obtained through microscope and Hoechst 33258 DNA staining method.Cell injury was assessed by CCK-8 methods.The levels of malonaldehyde（MDA）and hydroxyl radical were determined by biochemistry.The change of calcium signal was detected by microfluorescent technique.Results：With the concentration of glutamate administered to astrocytes increased,the injury of cell were severe more.Compare to control,the morphology of astrocytes pretreated by deferroxamine kept better to 0.5,1.0,5.0 mmol/L glutamate injury,cell viability（%）was increased（94.72±2.20,82.49±1.94,67.94±5.55,respectively,P0.05）,the ratio（%）of condensed nuclei decreased（10.44±5.03,7.93±5.04,11.01±3.73,respectively,P0.05）.The levels of hydroxyl radical and MDA decreased（10.17±1.79,7.36±1.47,respectively,P0.05）.Deferroxamine inhibited the increasing of Ca2＋ induced by glutamate significantly.Conclusion：Deferroxamine can protect injured astrocytes induced by glutamate.The protective activities of deferroxamine may be related to its ability to reduce Ca2＋ overload and the levels of MDA and hydroxyl radicals induced by glutamate.