谷氨酸对视网膜内层神经细胞影响的研究  

The experimental study of glutamate neurotoxicity on inner stratum retinal neurons

作　　者：钟一声[1] 叶纹[1] 王康孙[1] 

机构地区：[1]上海第二医科大学附属瑞金医院眼科,200025

出　　处：《中华眼底病杂志》2001年第4期311-314,共4页Chinese Journal of Ocular Fundus Diseases

摘　　要：目的　探讨谷氨酸对视网膜内层神经细胞 (inner stratum retinal neurons,ISRN)的影响及其毒性作用的量效关系。　方法　将 30只新生小鼠神经层视网膜组织块均匀接种于 2块 2 4孔培养板 (4 8孔 ) ,将其分成正常对照组、谷氨酸作用 2 4h组和正常培养 18h后 ,谷氨酸继续作用 6 h组。培养 2 4h后 ,将视网膜组织块常规进行切片 ,HE染色 ,油镜下观察视网膜神经节细胞层 (retinal ganglion cells,RGCs)和内核层 (inner nuclear layer,INL)的细胞形态 ,并分别计算细胞形态正常的 RGCs数和 INL 细胞数。　结果　视网膜组织正常培养 2 4h后 ,ISRN(RGCs和 INL细胞 )中可见少数细胞胞核固缩和坏死。谷氨酸作用 2 4h组 :当谷氨酸浓度为 2、4mmol时 ,ISRN的细胞形态与对照组之间差异无显著性意义 (P>0 .0 5 ) ;谷氨酸浓度≥ 6 mmol,视网膜内层形态正常的神经细胞数量明显较对照组减少 (P<0 .0 5 ) ,且随着谷氨酸浓度的增加 ,其数量逐渐减少。谷氨酸后 6 h作用组 :谷氨酸浓度为 6 mmol时 ,INL中形态正常的细胞数量明显较对照组减少 (P<0 .0 5 ) ,而 RGCs层中形态正常的细胞数量与对照组之间差异无显著性的意义 (P>0 .0 5 ) ;谷氨酸浓度≥ 8mmol时 ,RGCs层和 INL中形态正常的细胞数量均较对照组明显减少 (P<0 .0 5 )。?Objective To investigate the glutamate toxicity on inner stratum retinal neurons(ISRN) and the neurotoxicity quantity-efficacy relation. Method Retinal explants obtained from 30 neonatal mices were implanted into two pieces of 24-well culture plates (48 wells). The 48 wells were divided into three groups: control group, glutamate exposure 24 h group, and glutamate exposure with further lasting 6 h group. The retinal explants were sectioned, and then stained with HE after 24 h in vitro. The cells in retinal ganglion cells (RGCs) layer and inner nuclear layer (INL) were analyzed by light microscope at 1 000 × magnification , and the number of normal morphological cells was counted under three 1 000× magnification fields. Results Some cells in ISRN (include RGCs and INL cells) showed pyknotic nuclei and necrosis after 24 h in control culture. Glutamate exposure 24 h group:at the 2 mmol and 4 mmol concentrations of glutamate, the situation of the normal morphological cells in ISRN had no difference from that of the control group ( P >0.05). At the concentration of glutamate more than or equal to 6 mmol, the number of normal morphological cells in ISRN was significantly less than that of the control group ( P <0 05), and with the increase of glutamate concentration, the number of normal morphological cells was reduced. Glutamate exposure with further lasting 6 h group: at the concentration of glutamate equal to 6 mmol, the number of normal morphological cells in INL was significantly less than that of the control group ( P <0 05), while the number of normal morphological cells in RGCs layer had no difference between two groups ( P >0.05). At the concentration of glutamate more than or equal to 8 mmol, the number of normal morphological cells in RGCs layer and INL was significantly less than that of the control group ( P <0.05). Conclusion Glutamate has the neurotoxicity for ISRN in vitro, and the effect is dose-dependant.

关 键 词：谷氨酸 视神经生理学 神经元 视网膜内层神经细胞 

分 类 号：R774[医药卫生—眼科]