听源性惊厥大鼠惊厥后NMDA受体NR1亚基蛋白表达和功能研究  

Alteration in NMDA receptor subunit NR1 immunoreactivity and function in epilepsy-prone rat brain after audiogenic seizure

作　　者：罗强[1] 赵世刚[2] 吴希如[2] 

机构地区：[1]河南医科大学第一附属医院儿科,郑州450052 [2]北京医科大学第一医院儿科神经组,北京100034

出　　处：《河南医科大学学报》2001年第4期397-400,共4页Journal of Henan Medical University

基　　金：国家自然科学 (青年 )基金资助项目 3990 0 16 0

摘　　要：目的 :探讨惊厥后大脑皮质N 甲基 D 门冬氨酸 (NMDA)受体亚基NR1蛋白表达及其功能改变与癫痫发病机制的关系。方法 :P77PMC大鼠按惊厥后不同时间随机分为 6组 ,免疫组化每组 6个样本 ,配基结合实验每组 5个样本 ,制备大脑冰冻切片和大脑皮质突触膜。利用免疫组化技术 ,观察听源性癫痫易感大鼠P77PMC惊厥后不同时间NR1亚基蛋白的表达 ,应用配基结合实验观察听源性惊厥大鼠P77PMC惊厥后大脑皮质突触膜对3H MK80 1的结合改变 ,以及NMDA受体激动剂对其结合的影响。结果 :大脑皮质在惊厥后 4hNR1蛋白表达即开始增加 ,2 4～48h达高峰。反应体系中无任何NMDA受体激动剂时 ,惊厥后大脑皮质突触膜对 3H MK80 1的结合有增加趋势 ,但差异无显著性 (P >0 .0 5 ) ;当反应液中加入NMDA受体激动剂时 ,惊厥后大脑皮质突触膜对 3H MK80 1的结合明显增加 ,除惊厥后 4h与正常对照组比较差异无显著性外 ,其他惊厥后各时间点与正常对照组比较差异均有显著性(P <0 0 1)。结论 :听源性惊厥大鼠P77PMC惊厥后 ,大脑皮质NMDA受体NR1亚基蛋白表达增加 ,NMDA受体对其激动剂的敏感性增加 ,提示NR1亚基参与了P77PMC大鼠惊厥的发生与发展 ;Aim:To determine the protein expression of NR1 subunit and function change of NMDA receptor in cortex of epilepsy prone rat (P77PMC).Methods:Immunohistochemistry and radio ligand binding assay were respectively used to detect the protein expression of the NMDA receptor after audiogenic seizure and the binding of 3H MK801, the NMDA receptor open-channel blocker, to the rat cortex membrane. Results:The NR1 immunoreactivity increased time dependently and reached a peak at 24～48 h in cortex after audiogenic seizure.No statistically significant alterations were detected when none of glutamate,glycine or spermidine existed in the reaction system, but when the agonists existed in the reaction system, the increased sensitivity of 3H MK801 binding to the stimulatory action of glutamate, glycine and spermindine was significant. Conclusion: The protein expression of NR1 subunit and the function activity of the NMDA receptor increase after audiogenic seizure. Those changes may contribute to an altered excitability of neuronal network after audiogenic seizure, and suggest that the NR1 subunits participates in the generation and development of seizure in P77PMC rat.

关 键 词：癫痫 N-甲基-D-门冬氨酸受体 NR1亚基蛋白表达 配基结合实验 听源性惊厥 大鼠 

分 类 号：R742.1[医药卫生—神经病学与精神病学]