致痫灶靶向TD-PF胶束介导KCNJ4调控Kir2.3/Cavelion1共轭分子内向整流效应的研究

Refractory epilepsy pathogenesis is quite complex, the relevant ion channels mechanism is now becoming a hot spot. This study attempts to apply earlier building TD- PF active targeting polymer micelle as the carrier, carry the reducing expressed KCNJ4 gene in intractable epileptogenic zone then transfect it to model rats. By in vitro and in vivo experiments, we want to observe target gene expression and behavior of rats, brain electrical physiological and cell excitability as well as the potassium channel conductance changes, before and after KCNJ4 gene transfection. Combineding the technology of siRNA interference which stimulated the KCNJ4 genetic degradation as contrast experiments, we are expected to reveal that the abnormal expression and function changes of inward rectifier potassium channel gene KCNJ4 and its encoded Kir2.3 subunits may be the bases on epilepsy discharge cycle; we also validate the downstream Cavelion1 molecules within assisting Kir2.3 rectification, driving potassium ion of the conjugate adjustment. If we can confirm the correlation between Kir2.3 with intractable epilepsy and the synergistic effect between Kir2.3/ Cavelion 1 conjugate molecules, it may provide us a complete understanding of the inward rectifier mechanism of refractory epilepsy, then further offer experimental foundation and theoretical basis to conduct the KCNJ4 gene and protein Kir2.3 as the new ideal of antiepileptic drugs sites.

难治性癫痫发病机制复杂，与其相关的离子通道异常正成为热点。本研究尝试应用构建合成的TD-PF主动靶向聚合物胶束作为载体，将前期发现表达下调的KCNJ4基因靶向转染至模型大鼠脑内致痫灶，通过体内外实验观察干预前后目标基因表达及大鼠行为学、脑电生理与细胞兴奋性及钾通道电导变化，结合siRNA干扰技术激发KCNJ4降解进行对比实验，可望揭示内向整流钾通道KCNJ4基因及其编码的Kir2.3亚单位表达和功能异常可能是痫样放电循环产生的关键分子与生理学基础；同时验证通路下游Cavelion1分子在协助Kir2.3发挥内向整流效应、驱动钾离子内移时的共轭调节作用。如能明确Kir2.3与难治性癫痫发生间的相关性以及Kir2.3/ Cavelion1共轭分子间的协同效应，则可能为完整理解难治性癫痫的内向整流相关机制、进一步将KCNJ4及Kir2.3作为抗癫痫药物的新型理想作用位点，提供实验基础与理论依据。