Wang S, Lees G J, Rosengren L E, Karlsson J E, Stigbrand T, Hamberger A, Haglid K G
Institute of Neurobiology, University of Göteborg, Sweden.
Brain Res. 1991 Feb 15;541(2):334-41. doi: 10.1016/0006-8993(91)91034-x.
The study employed an immunochemical quantification of brain cell marker proteins in addition to quantitative morphology in order to provide a more multifacetted and characterized model for an excitotoxic CNS lesion. The importance of the approach in the evaluation of the potential of neuroprotective agents is emphasized. The S-100 protein, the glial fibrillary acidic (GFA) protein, neuron specific enolase (NSE) and neuronal intermediary filament polypeptides (NF 68 and NF 200) were measured with a dot-immunobinding assay, 3-30 days after a unilateral injection of N-methyl-D-aspartate (NMDA) in the left dorsal hippocampus of the rat. After 3 days, the neuronal cell loss averaged 80% in the hippocampus. The S-100 content was reduced 3 days after injection, but was 150% of control at 30 days. GFA increased constantly from days 3 to 30. The neuronal marker proteins were all markedly reduced 7 days after injection. However, at 30 days, NF 68 and NF 200 were close to control (80%). Increasing content would reflect regeneration and sprouting of neurites. The content of the neuronal cytoplasmic marker, NSE, was significantly lower than control also at 10 and 30 days, although a gradual recovery could be traced.
该研究除了进行定量形态学分析外，还采用免疫化学方法对脑细胞标记蛋白进行定量，以便为兴奋性毒性中枢神经系统损伤提供一个更具多面性和特征性的模型。强调了该方法在评估神经保护剂潜力方面的重要性。在大鼠左背侧海马体单侧注射N-甲基-D-天冬氨酸（NMDA）后3 - 30天，采用斑点免疫结合测定法测量S-100蛋白、胶质纤维酸性蛋白（GFA）、神经元特异性烯醇化酶（NSE）和神经元中间丝多肽（NF 68和NF 200）。注射3天后，海马体中神经元细胞损失平均达80%。注射后3天S-100含量降低，但在30天时为对照的150%。从第3天到第30天，GFA持续增加。注射7天后，所有神经元标记蛋白均显著降低。然而，在30天时，NF 68和NF 200接近对照水平（80%）。含量增加反映神经突的再生和出芽。尽管可以追踪到神经元胞质标记物NSE的含量逐渐恢复，但在10天和30天时仍显著低于对照。
