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目的:探讨谷氨酸对体外原代培养的大鼠耳蜗螺旋神经节细胞(SGC)兴奋性毒性作用。方法:SGC在体外培养4d,加入1mmol/L谷氨酸,继续培养24h后,用荧光显微镜观察Annexin-V-FITC和PI双标的细胞形态改变,以及用激光共聚焦方法观察细胞内游离钙离子浓度的变化。结果:体外培养4d的SGC,用1mmol/L谷氨酸处理24h后,与对照组相比,出现大量的细胞受损,表现为死亡或凋亡(P<0.05);用Fluo-3染色,激光共聚焦显微镜观察,加入谷氨酸后,80%的SGC细胞内钙离子迅速增加,150s达到高峰。结论:谷氨酸诱导的SGC钙离子内流与细胞受损有明显的关系。
Objective: To investigate the excitotoxicity of glutamate on cultured primary cultured rat cochlear spiral ganglion cells (SGC). Methods: SGC cultured in vitro for 4 days, adding 1mmol / L glutamic acid, cultured 24h later, observed by fluorescence microscopy Annexin-V-FITC and PI double-labeled cells morphological changes, and confocal laser scanning confocal microscope to detect intracellular free calcium Changes in concentration. Results: After treated with 1mmol / L glutamic acid for 24 hours, a large number of cell damage occurred in SGC cultured in vitro for 4 days, showing death or apoptosis (P <0.05). Fluo-3 staining, Confocal laser scanning microscopy, adding glutamate, 80% of the SGC intracellular calcium increased rapidly, 150s peaked. CONCLUSION: Glutamate induced SGC Ca2 + influx has a clear relationship with cell damage.