全蝎提取物BmP02通过调控小胶质细胞钾通道Kv1.3抗癫痫发生的分子机制

颞叶癫痫发生的细胞信号机制尚未明了，临床治疗效果有限。最新资料提示钾通道Kv1.3的功能异常会引起小胶质细胞钙稳态失衡，诱发小胶质细胞自噬障碍和神经炎症，参与癫痫发生。中医临床实践证明全蝎熄风止痉的功效显著，抗癫痫效果明显。申请人在青年基金的研究中提取了其中13种活性成分，发现组分BmP02可阻断Kv1.3，有促进小胶质细胞自噬，缓解小鼠癫痫发生的作用。故推测BmP02可通过抑制Kv1.3异化小胶质细胞胞内钙信号，减轻由小胶质细胞介导的神经炎症，干预癫痫病状。为此，本申项拟利用Kv1.3敲除小鼠结合电生理学、双光子成像、单细胞测序及结构生物学等手段，探讨⑴BmP02干预小鼠颞叶癫痫行为与脑电的效应，对神经炎症的缓解作用；⑵BmP02识别Kv1.3的分子结构信息；⑶BmP02调控小胶质细胞自噬、炎症途径的信号整合机理。为解析全蝎熄风止痉的功效提供理论依据，为抗癫痫药物设计贡献可能的分子模板。

The cellular signaling mechanism of temporal lobe epilepsy is still unclear, and its clinical therapeutic effects are often limited. The recent studies suggested that the dysfunction of potassium channel Kv1.3 leads to the imbalance of calcium homeostasis in microglial cells, which induces microglial cell autophagic dysfunction as well as neuroinflammation, and mediates epileptogenesis. The clinical practice of traditional Chinese medicine proves that scorpion has significant efficacy of "Xi Feng Zhi Jing" and antiepileptic effects. The applicant extracted 13 active ingredients from scorpion during the research of youth fund, and found that BmP02 could block Kv1.3 channels, promoting microglia cell autophagy and alleviating epilepsy in mice. It is suggested that BmP02 could interfere with epilepsy and suppress the neuroinflammatory pathway by inhibiting microglial Kv1.3 and modulating intracellular calcium signals. Therefore, this project aims to use Kv1.3 knockout mice combined with electrophysiology, two-photon imaging, single cell sequencing, structural biology and other techniques, to illuminate (1) the alleviating effects of BmP02 on temporal lobe epilepsy behavior, electroencephalogram and neuroinflammation in mice; (2) the molecular structure information of BmP02 recognizing the Kv1.3; (3) the integration mechanism of BmP02 regulating microglial cell autophagy and inflammatory signals. It provides a theoretical basis for clarifying the scorpion's efficacy of "Xi Feng Zhi Jing", and contribute a possible molecular template for the design of antiepileptic drugs.