丙戊酸钠处理对于癫痫大鼠模型海马内细胞自噬的影响

Influence of sodium valproate on autophagy in the hippocampus of epilepsy rat model

目的:研究丙戊酸钠(VPA)对癫痫模型大鼠海马内细胞自噬、动物脑电图和行为学的影响。方法:制作癫痫大鼠模型并随机分为癫痫组、3甲基腺嘌呤(3MA)组、VPA组及VPA联合3MA组四组,设正常大鼠为对照组。观察各组行为学及脑电图变化,HE及Nissl染色显示神经元的损伤情况,免疫组织化学染色、Western Blot、Real-time PCR检测自噬标记物微管相关蛋白1轻链3(LC3)表达。结果:行为学观察及脑电图检测显示,丙戊酸钠组及丙戊酸钠联合3MA组均可降低癫痫发作等级,3MA组无抗癫痫效果。HE及Nissl染色显示,对照组未见明显异常,癫痫组神经元损伤严重,丙戊酸钠组与癫痫组相比,神经元损伤明显减轻;3MA组与癫痫组相比,神经元损伤明显减轻;丙戊酸钠联合3MA组与丙戊酸钠组相比,神经元损伤明显减轻。免疫组织化学染色、Western Blot、Real-time PCR检测显示,对照组LC3呈低表达,癫痫组LC3表达明显高于对照组;丙戊酸钠组LC3表达明显高于癫痫组;3MA组LC3表达明显低于癫痫组;丙戊酸钠联合3MA组LC3表达明显低于丙戊酸钠组。结论:VPA处理能够诱导癫痫大鼠海马内细胞自噬增加和导致神经元损伤。

Objective： To investigate the effect of sodium valproate（VPA） on autophagy,electroencephalogram and behavior in the hippocampus of rats with epilepsy. Methods： Epileptic rat model were established and the kindled rats were randomly grouped into four groups,named seizure group,3 methyladenine（ 3MA） group,VPA group and VPA ＋3MA group,respectively. Normal rat were named control group. The behavior and electroencephalogram of rats were observed and recorded. The neuronal damage was observed by HE and Nissl staining. And the levels of microtubule-associated protein light chain 3（ LC3） was tested by immunohistochemistry staining,Western Blot and Real-time PCR.Results： The changes of behavior and electroencephalogram indicated that VPA and VPA ＋ 3MA can reduce the seizure intensity effectively,but 3MA can＇t. The HE and Nissl staining results show that there is no obviously abnormal change in control group,the neuronal damage in seizure group was severe,the neuronal damage in VPA group was significantly attenuated compared to seizure group,the neuronal damage in 3MA group was significantly attenuated compared to seizure group and the neuronal damage in VPA ＋ 3MA group was significantly attenuated compared to VPA group. The results of immunohistochemistry,Western Blot and Real-time PCR indicated that expression level of LC3 in control group was low,expression level of LC3 in seizure group was significantly higher than control group and the expression level of LC3 in VPAgroup was significantly higher than seizure group,the expression level of LC3 in 3MA group was significantly lower than seizure group and the expression level of LC3 in VPA ＋ 3MA group was significantly lower than VPA group. Conclusion：VPA could elevate the level of autophagy in hippocampal neurons of epileptic rat model and resulted in neuronal damage.