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目的 :研究小鼠持续性脑缺血后不同脑区 NMDA受体亚单位表达的变化及其与病理损伤之间的联系。方法 :采用大脑中动脉阻断法制作小鼠持续性脑缺血模型 ,取缺血后不同时间的皮层、海马、皮层下脑组织 ,用免疫印迹技术测定 NMDA受体亚单位 ζ1和 ε1及 ε2蛋白的含量。同时作冰冻切片和苏木素伊红染色 ,计算相应脑区神经元密度。结果 :NMDA受体 ζ1和 ε1及 ε2亚单位表达的改变发生于缺血后 5 h内 ,皮层下 3个亚单位表达均明显增加 ,海马则各亚单位表达变化不一。脑梗塞灶和相关脑区神经元密度显著减小均出现于缺血后 5 h,相关脑区神经元死亡严重程度依次为皮层下组织和海马 CA1区及大脑颞叶皮层 - 层。结论 :小鼠持续性脑缺血 ,缺血侧脑组织 NMDA受体亚单位 ζ1和 ε1及 ε2表达的变化发生在明显的病理损害之前 ,表达变化程度因脑区不同 ,与相关脑区神经元损害的程度相应。
Objective: To investigate the changes of NMDA receptor subunits in different brain regions and their relationship with pathological injury after chronic cerebral ischemia in mice. Methods: The model of persistent cerebral ischemia was established by middle cerebral artery occlusion (MCAO). The cortex, hippocampus and subcortex brain tissue were taken at different time after ischemia. The ζ1, ε1 and ε2 of NMDA receptor subunits were determined by Western blotting. Protein content. At the same time for frozen sections and hematoxylin eosin staining, calculate the corresponding brain area neuron density. Results: The expression changes of ζ1, ε1 and ε2 subunits of NMDA receptors occurred within 5 h after ischemia, the expressions of three subunits in the subcortical layers were significantly increased, while the expressions of subunits in the hippocampus varied. Cerebral infarction and related brain regions neuronal density were significantly reduced in 5 h after ischemia, the severity of neuronal death in the relevant brain regions were subepithelial tissue and hippocampal CA1 and cerebral temporal cortex - layer. CONCLUSION: The changes of the expression of ζ1, ε1 and ε2 of NMDA receptor subunits of ischemic brain tissue in mice with persistent cerebral ischemia occurred before the obvious pathological changes. The changes of the expression level were related to the different brain regions, The degree of damage accordingly.