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本研究用戊四氮(pentylenetetrazole,PTZ)方法建立了一种斑马鱼幼体癫痫模型。结果显示,PTZ可导致斑马鱼幼体癫痫样异常兴奋行为,如游动速度加快、抽搐等异常现象;癫痫标志基因c-fos信号在斑马鱼幼体脑区增强并范围扩大,而lgi1基因表达则被抑制。这些表型与癫痫临床病例和文献报道相符。给予癫痫治疗药丙戊酸钠(VPA)可使斑马鱼癫痫症状减轻或消失:异常兴奋游动、c-fos和lgi1基因表达,以及神经核蛋白Neu N均恢复到近正常组水平。利用所建模型,检测了两种先导化合物Y53和BMT(均为小檗碱结构类似物)的抗癫痫作用,Y53抑制光刺激下的发作更有效;BMT对无刺激的发作有较好的抑制作用。综上,所建斑马鱼癫痫模型对于化合物有无刺激因素条件下的差异作用也具有较好的辨识力,可作为抗癫痫先导化合物的简便实用的药效筛选平台及用于癫痫发病机制研究。
In this study, a pentylenetetrazole (PTZ) method was used to establish a zebrafish larval epilepsy model. The results showed that PTZ could induce abnormal epileptiform activity in zebrafish larvae such as accelerated swimming and convulsions. The c-fos signal of epilepsy marker increased and expanded in the brain of zebrafish larvae, while the expression of lgi1 gene was inhibition. These phenotypes are in line with clinical cases of epilepsy and reported in the literature. Epilepsy treatment with sodium valproate (VPA) reduced or eliminated the symptoms of zebrafish epilepsy: abnormal excitement, c-fos and lgi1 gene expression, and neuronal protein Neu N returned to nearly normal levels. Using the established model, the antiepileptic effects of two lead compounds, Y53 and BMT (both are structural analogues of berberine) were examined, and Y53 was more effective in suppressing the onset of light-stimulated events; BMT was more effective in suppressing non-stimulated seizures effect. In summary, the model of zebrafish epilepsy also has good discriminating ability for compound under the condition of stimulating factors, which can be used as a simple and practical drug screening platform for anti-epileptic lead compounds and for studying the pathogenesis of epilepsy.