N-甲基-D-天冬氨酸及其非竞争性受体阻滞剂和钙离子对培养的鼠视网膜神经细胞的作用  

The influencing actions of NMDA, MK801 and Ca^(2+) on cultured retinal neuron cells

作　　者：刘海泉[1] 葛坚[1] 杨智宽[1] 卓业鸿[1] 林明楷[1] 

机构地区：[1]中山大学中山眼科中心,广州510060

出　　处：《中华眼科杂志》2002年第3期148-150,共3页Chinese Journal of Ophthalmology

基　　金：国家自然科学基金资助项目 (3 9770 789) ;广东省卫生厅基金资助项目 (B19970 3 3 )

摘　　要：目的　探讨N 甲基 D 天冬氨酸 (N methyl D aspartate,NMDA)及其非竞争性受体阻滞剂(MK80 1)和Ca2 + 对培养的鼠视网膜神经细胞的作用及其相互间的关系。方法　取生后 1～ 3d鼠龄的Sprague Dawley(SD)乳鼠视网膜作为原代视网膜神经细胞进行体外培养 ,对培养 7d的视网膜神经细胞经抗神经丝蛋白 (neurofilamentprotein ,NF)、Thy1 1及抗胶质纤维酸性蛋白 (glialfibrillaryacidicprotein ,GFAP)单克隆抗体进行鉴定 ,同时在培养 7d的神经细胞中单独及联合加入NMDA、MK80 1和Ca2 + ,锥虫蓝拒染 ,计算细胞活性。结果　 2 0 0 μmol/L的NMDA及 10mmol/L的Ca2 + 所致视网膜神经细胞失活与对照组比较 ,差异有显著意义 (F =2 6 0 91;P =0 0 2 5 ,P <0 0 0 1) ,2 0 μmol/L的MK80 1对视网膜神经细胞的影响不明显 ,对Ca2 + 所致细胞毒性无保护作用 ,但可阻断NMDA的作用。结论NMDA、一定浓度的Ca2 + 对培养的视网膜神经细胞有毒性作用 ,MK80 1可阻断NMDA的毒性作用 ,但对Ca2 + 的毒性无阻断作用 ,Ca2 +Objective To investigate the effects of N methyl D aspartate (NMDA), MK801 (non competitive blocking agent of NMDA receptor) and Ca 2+ on cultured retinal neurons and the interaction of these reagents. Methods NMDA, MK801 and Ca 2+ were singly and jointly added to the retinal neurons after primary culture for 7 days which obtained from 1 3 day old postnatal Sprague Dawley rat. The cells were identified with anti neurofilament protein (NF), Thy1.1 and anti glial fibrillary acidic protein (GFAP) monoclonal anti body. The effects were assessed quantitatively using the trypan blue exclusion methods. Results Neuronal death which induced by 200 μmol/L NMDA and 10 mmol/L Ca 2+ was significantly different from that of the control, 20 μmol/L MK801 had no significant effect on cultured retinal neurons, and did not protect the cell from neurotoxicity of Ca 2+ , but could block the neurotoxicty of NMDA. Conclusions NMDA and certain concentration of Ca 2+ are neurotoxic to retinal neurons in the primary cultured. MK801 can block the neurotoxicity of NMDA, but not the neurotoxicity of Ca 2+ . Neuronal death induced by Ca 2+ may get through another mechanism.

关 键 词：视网膜 受体 N-甲基-D-天冬氨酸 MK801 钙 视网膜神经细胞 阻滞剂 动物实验 

分 类 号：R770.2[医药卫生—眼科]