丙戊酸钠对大鼠海马神经元癫痫样放电后P-CREB1表达的影响

Effects of valproate sodium on phosphorylatted cAMP responsive element binding protein after hippocampal neuronal epileptiform discharge in rat

目的:研究丙戊酸钠对大鼠海马神经元癫痫样放电后磷酸化腺感苷反应元件结合蛋白1(Phosphorylated cAMP responsive element binding protein 1,P-CREB1)表达的影响。方法:wistar新生鼠,迅速断头取脑,体外培养海马神经元,建立神经元癫痫样放电模型,将神经元分为空白组、模型组、丙戊酸钠低剂量(50mg/L)组、丙戊酸钠高剂量(100mg/L)组,运用免疫荧光技术观察P-CREB1在神经元癫痫样放电后在细胞内的表达部位,采用wester blot技术测定P-CREB1在不同分组中的表达强度。结果:通过免疫荧光技术,在各组中都可以看到P-CREB1在细胞核内表达,以模型组最明显;运用Western blot,发现表达趋势与免疫荧光一致,并且,给予丙戊酸钠后,P-CREB1表达减弱,且高剂量组与低剂量组之间差异有统计学意义(P<0.05)。结论:海马神经元无镁处理后呈癫痫样放电,同时P-CREB1被过度激活,而有效浓度的丙戊酸钠可抑制此反应P-CREB1的磷酸化水平。

Objective：To investigate the effects of valproate sodium on P-CREB1 after hippocampal neuronal epileptiform discharge in rat.Methods：The neonate wistar rats were decapitated quickly to obtain the hippocampal neuron,Which were cultured in vitro,After the epileptiform discharge model of neuron was established,neurons were divided into control group,model group,low valproate dose group（50mg/L） and high dose group（100mg/L）.Expression positions of P-CREB1 after epileptiform discharge were examined by immunofluorescence technique,and Western blot was used to examine the expression intensity of P-CREB1 in different group.Results：Through immunofluorescence,P-CREB1 was observed in the nucleus of each group,and the most intensive expression was found in model group.Through western blot,the expression tendency was found to be the same as the former Results,Moreover,after added valproate sodium,the expression of P-CREB1 decreased,and there was statistical significance of the difference between low valproate dose and high dose（P〈0.05）.Conclusion：P-CREB1 could be persistently activated in the hippocampal neuron,which presents epileptiform discharge.Valproate sodium could decrease the phosphorylation of P-CREB1 in effective dose.