发育中大鼠皮层神经元惊厥样放电中PSD-95 mRNA表达改变

Effects of recurrent epileptiform discharges induced by magnesium-free treatment on alternations of PSD-95 mRNA expression in developing cortical neurons of rats

目的探讨早期无镁细胞外液处理后惊厥样放电对发育中大鼠皮层神经元PSD-95 mRNA表达的影响。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Neurobasal/B27培养液组(CONT组)、正常细胞外液组(PS组)和无镁细胞外液组(MGF组)。神经元分别在上述三种液体中孵育3h,然后换为正常Neurobasal/B27培养液继续培养,在处理后6,24,72h及处理后第6天时应用实时定量PCR测定PSD-95 mRNA的表达。结果与CONT组和PS组相比,MGF组PSD-95 mRNA表达在处理后24h明显增高(P<0.05)。处理后6h,72h及第6天的皮层神经元PSD-95 mRNA表达三组中两两比较差异无统计学意义(P>0.05)。结论发育中大鼠皮层神经元惊厥样放电可以导致神经元PSD-95 mRNA表达的改变。

Objective To explore the changes of PSD-95 mRNA expression induced by early magnesium-free treatment in developing cortical neurons of rats. Methods The seizure model of recurrent epileptiform discharges was induced by transient magnesium-free media treatment in primary cultured embryo cortical neurons of rats. According to different treatments to cortical neurons at culture day 6,the neurons were divided into 3 groups： regular Neurobasal/B27 medium group（ control group） ,regular extracellular solution medium group（ PS group） ,and magnesium-free medium group（ MGF group） . The neurons were exposed to the above three media for 3 h at culture day 6,and then cultured in regular Neurobasal/B27 medium again. The expression of NMDAR subunit of PSD-95 mRNA was measured using fluorescent real-time quantitative PCR at 6 h,24 h,72 h and 6 d after treatment. Results Compared with control and PS groups respectively,the expression of PSD-95 mRNA increased in MGF group at 24 h（ P 0. 05） . There was no statistical difference among three groups at 6 h,72 h and 6 d （ P 0. 05） . Conclusion The recurrent epileptiform discharges induced by magnesium-free treatment could induce the changes of PSD-95 mRNA expression in developing cortical neurons of rats.