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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 + 致神经元死亡可能还有其他途径
Objective To investigate the effects of N-methyl D aspartate (NMDA) and its noncompetitive receptor blockers (MK80 1) and Ca2 + on cultured rat retinal neurons and their relationship with each other . Methods Retinal neurons of Sprague Dawley (SD) rats aged 1 ~ 3 days after their birth were cultured as primary retinal neurons in vitro. The retinal neurons cultured for 7 days were stimulated with anti-neurofilament protein (NF), Thy1 1 Glial fibrillary acidic protein (glialfibrillaryacidicprotein, GFAP) monoclonal antibody identified at the same time in the cultured 7d nerve cells alone and in combination NMDA, MK801 and Ca2 +, trypan blue exclusion, calculate cell activity. Results The inactivation of retinal neurons induced by 200 μmol / L NMDA and 10 mmol / L Ca2 + was significantly different from that of the control group (F = 26.01 1, P = 0 025, P <0 The effect of MK80 1 at 20 μmol / L on retinal neurons was not obvious, and there was no protective effect on the cytotoxicity induced by Ca2 +, but it could block the effect of NMDA. Conclusion NMDA, a certain concentration of Ca2 + has a toxic effect on cultured retinal neurons. MK80 1 can block the toxic effect of NMDA, but it has no effect on the toxicity of Ca2 +, and there may be other ways of Ca2 + -dependent neuronal death