早期惊厥样放电对发育中神经元NMDA受体1表达的远期影响(英文)  

Long-term effect of early discharges on NMDAR-1 expression in developing neurons

作　　者：王静敏[1] 姜玉武[1] 姜茜[1] 曹海燕[1] 倪宏[1] 薄涛[1] 吴希如[1] 

机构地区：[1]北京大学第一医院儿科,北京100034

出　　处：《山西医科大学学报》2007年第2期97-102,共6页Journal of Shanxi Medical University

基　　金：国家自然科学基金资助项目(30270448,30470555);北京大学国家"211工程"资助项目;第三十三届博士后科学基金资助项目(2003033082)

摘　　要：目的探讨早期惊厥样放电对发育中大鼠皮层神经元N-甲基-D-天冬氨酸受体1(N-methyl-D-aspartatere-ceptor1,NR1)表达的远期影响。材料与方法以原代培养的大鼠大脑皮层神经元无镁诱导的反复惊厥样放电为模型。根据对培养6d皮层神经元的不同处理分为三组:正常DMEM培养液组(CONT1)、正常细胞外液组(CONT2)和无镁细胞外液组(MGF)。神经元分别在上述三种液体中孵育3h,然后恢复正常DMEM培养基继续培养,分别应用实时定量PCR与West-ernblot方法在培养7、12及17d时测定皮层神经元NMDA受体1mRNA与蛋白表达的变化。结果在正常DMEM培养液中,NR1mRNA与蛋白的表达在培养7d时处于较高水平,以后逐渐下降,以培养12d为最低。与CONT2和CONT1组相比,MGF组NR1mRNA与蛋白的表达在培养7d皮层神经元均明显降低,在12d时明显增高(P<0.05)。CONT2组与CONT1组相比无统计学差异(P>0.05)。结论发育中皮层神经元早期无镁诱导惊厥样放电对NMDA受体1mRNA与蛋白表达具有远期影响,提供了神经元早期惊厥样放电引起远期功能损伤的可能机制。Objective To explore the long-term effect of early discharges on NMDAR-1 expression in developing rat cortical neurons. Methods The seizure model was recurrent epileptiform discharges induced by transient magnesium-free media treatment in the primary cultured embryo cortical neurons of rat. According to different treatment to cortical neurons at cultured day 6, ceUs were assigned to three groups respectively based on the mediums they were transiendy exposed to. The neurons were exposed to Dulbecco＇ s Modified Eagle＇s Medium （DMEM） in control group 1 （CONT1）,exposed to a physiological solution （PS） in control group 2 （CONT2） ,and exposed to the same medium as CONT2 except for the removal of magnesium in the magnesium-free physiological solution （MGF） group. Following a 3 h exposure, the expression of N-methyl-D-aspartate receptor 1 （NR1） mRNA and protein were measured at different culture days （7,12,17） using real-time PCR and Westem blot assays. Results The expression of NR1 mRNA and protein in DMEM medium had higher value level at culture day in vitro （DIV） 7, and then they decreased gradually and had the lowest value at DIV12. They increased again with development. Compared with groups CONT1 and CONT2 respectively,expression of NRI mRNA and protein in group MGF decreased at DIV7 and increased at DIV12 （P〈0.05） ,and there were no statistic significantdifferencesatDIV17. Conclus/on The results suggest that NRl mRNA and protein rmy alter with the devetopment and there are long-term effects of Mg^2＋ -free induced seizure-llke discharges on expression of NR1 mRNA and protein in devdeping rat cortical neurons.

关 键 词：发作 受体 N-甲基-D-天冬氨酸 大鼠 

分 类 号：R338.8[医药卫生—人体生理学]