FBP1通过调控神经元糖代谢和兴奋性在颞叶癫痫发病中的作用和机制研究

Epilepsy is defined as a chronic central nervous system disease due to abnormal excessive and synchronous neuronal activity in the brain. At present, treatment efficacy based on antiepileptic is still unsatisfying, possibly for the unclear pathogenesis. Interictal FDG PET showed decreased glucose metabolism of nervous in epileptic foci. However, the cause and relationship between the electrophysiological disorder and glucose hypometabolism of neuron have still not been clarified. We found the increased expression of fructose-1,6-bisphosphatase 1 (FBP1), accompanied by the decreased glucose transport and phosphorylation in epileptic focus. Another study demonstrated that the substrate of FBP1 could down-regulate expression of γ-aminobutyric acid A receptor (GABAAR), which subsequently enhanced the intrinsic excitability and seizure susceptibility of neurons. Therefore, we speculate that FBP1 might be involved in reducing glucose metabolism, simultaneously increasing neural excitability in epileptic foci, eventually promoting the occurrence of epilepsy. This study intends to investigate the role and mechanism of FBP1 in regulating the decrease of glucose metabolism and the increase of nerve excitability by using the epileptic mouse model, and to clarify the relationship of FBP1, key enzymes of glucose metabolism pathway and GABAAR. Finally, the model of infecting organotypic hippocampus slice, changing of FBP1 expression and microPET will be utilized to verify. The purpose of this study are to conduct inquiries into the role and mechanism of FBP1 in epilepsy from the point of view of glucose metabolism and neuroelectrophysiology, and to provide a new theoretical basis and target for the treatment of epilepsy.

癫痫是临床常见的以脑内神经元（NC）反复异常同步放电引起的中枢神经功能障碍，其发病机制仍未完全明确，抗癫痫治疗效果不理想。癫痫灶发作间期FDG PET表现为糖代谢减低，但究其原因及与NC电生理紊乱之间相互关系尚未阐明。前期我们研究发现癫痫灶NC内果糖-1,6-二磷酸酶1（FBP1）表达增高，葡萄糖转运和磷酸化过程减弱。另有研究发现FBP1的底物可下调γ-氨基丁酸A型受体（GABAAR）的表达，使NC兴奋性增加。据此我们推测FBP1可能同时参与对NC糖代谢和兴奋性的双向调控，促进癫痫发生发展。本研究拟通过癫痫鼠模型首次探讨FBP1调控NC糖代谢减低和兴奋性增加的作用及机制，明确FBP1与糖代谢路径关键酶及GABAAR之间的关系。随后，将借助海马脑片模型、FBP1的干预和PET显像进行验证。该研究旨在从糖代谢和电生理角度探讨FBP1在癫痫中的作用及机制，为癫痫治疗提供全新的靶点和理论依据。

癫痫是临床常见的以脑内神经元反复异常同步放电引起的中枢神经功能障碍，经临床合理、规范的抗癫痫药物治疗后，但仍有三分之一的患者会发展为药物难治性癫痫，针对其发病机制仍未完全明确，抗癫痫治疗效果不理想临床科学问题，根据前期研究结果推测癫痫灶内果糖-1,6-二磷酸酶1（FBP1）表达增高同时参与了神经元糖代谢降低和兴奋性增高的双向联合调控，促进癫痫发生发展。本项目原拟通过癫痫患者术后癫痫灶组织、颞叶癫痫鼠模型探讨FBP1调控海马区神经元糖代谢减低和兴奋性增加的作用及机制，首先明确FBP1与糖代谢路径关键酶（GLUT-3/HK)、低氧诱导因子（HIF-1A）及γ-氨基丁酸A型受体（GABAAR）之间的关系。随后，拟借助海马脑片模型病毒转染、癫痫鼠模型海马区脑立体定性注射病毒两种方式对FBP1表达进行干预，通过多靶点PET显像和组织学检测进行验证，旨在从糖代谢和电生理角度探讨FBP1在难治性癫痫发生发展中的作用及机制，为癫痫的防治提供新的思路。本项目首先利用动态18F-FDG PET分析明确了难治性癫痫患者致痫灶主要存在葡萄糖磷酸化过程减低，然后成功构建癫痫大鼠模型进行18F-FDG/18F-SynVesT-1 microPET显像及海马区上述指标的组织检测，基本明确了FBP1在癫痫组大鼠中的表达量高于同时期正常组大鼠，GABAAR和GLUT-3在癫痫组大鼠中的表达量低于同时期正常组大鼠，说明FBP1同时参与调控神经元糖代谢与兴奋性机制和内在联系，为未来癫痫治疗提供新的线索。研究者随后开展了基于颞叶癫痫患者18F-FDG PET脑显像SPM分析、手术目标脑区多模态影像的深度学习、突触密度18F‑SynVesT‑1 PET显像对癫痫灶病因和定位诊断、颞叶癫痫脑网络分析及其机制探讨，为临床在癫痫病因和致痫灶定位诊断、难治性癫痫术前评估和预后评价提供了重要的技术手段和数据支撑，具有良好的临床应用前景；同时为FBP1癫痫灶糖代谢-电生理紊乱机制在癫痫发生发展中的作用及其机制提供实验室依据，为癫痫防治提供全新的靶点和理论依据。