Sv2A转基因干细胞移植在难治性癫痫治疗中的应用研究

Recent studies of the antiepileptic drug levetiracetam (LEV) have revealed that the reduced expression of synaptic vesicle protein 2A (SV2A), the binding target of LEV, was closely related with the onset of epilepsy. Such findings showed that SV2A may be a new target for epilepsy treatment. This research project will study the efficacy and mechanism of SV2A's antiepileptic property via SV2A expression using a lentiviral bicistronic expression vector (Lenti-EBCL2,-SV2A). On the other hand, the green fluorescent protein (EGFP) fused anti-apoptotic BCL2 gene will be used to increase the survival of the transplanted cells. In this research, we will use several methodologies, including the behavior, electrophysiology, and pathology, to study the treatment of refractory epilepsy using transgenic cell transplantation. At the same time, we will analyze the survival and differentiation of implanted cells and the regeneration of nerve fibers using Feridex labeling, as well as molecular biology and immunological assays to measure GFAP, of NF200 and NSE. We believe that our research will further improve the current understanding of the impact of SV2A in the treatment of epilepsy and its mechanism. In addition, this project will help promote the application of stem cell transplantation in the treatment of epilepsy and lay an important experimental foundation for future clinical research on this topic.

抗癫痫药物左乙拉西坦(Levetiracetam,LEV)的研究发现其药物结合靶点突触囊泡蛋白2A(synaptic vesicle protein2A,SV2A)的表达降低与癫痫的发作具有密切的相关性。特别是癫痫持续状态SV2A在海马硬化的颞叶癫痫患者中表达下降更为明显。提示SV2A有可能成为癫痫治疗新的靶点。但是转基因干细胞在中枢神经系统微环境中存活率是影响干细胞移植治疗效果的关键因素。因此本课题在研究SV2A转基因干细胞对癫痫治疗作用的同时引入BCL-2抗细胞凋亡的研究方向。在研究SV2A对癫痫治疗作用的基础上，拟用慢病毒载体构建Lenti-EBCL2-SV2A表达载体，筛选和建立稳定表达SV2A和BCL2干细胞。并进一步研究SV2A在癫痫治疗中的作用机制和移植干细胞的存活、迁移、分化和宿主整合。为转基因细胞移植在癫痫临床治疗中的应用奠定实验基础。

癫痫（Epilepsy）是一种脑神经功能障碍性疾病综合征，世界范围内常见的神经系统疾病之一。本次研究中采用戊四氮（PTZ）亚惊厥剂量腹腔注射建立慢性癫痫模型，造模成功后予以白藜芦醇（Res）干预，观察大鼠的癫痫发作行为学的变化，行Morris水迷宫实验测试大鼠的空间学习和记忆能力，并检测大鼠脑脊液、血清S100B蛋白的影响，发现白藜芦醇能够改善癫痫大鼠的发作时间和发作级别，能够改善空间学习记忆能力，并且能够降低 PTZ致痫大鼠脑脊液、血清S100B蛋白含量(p<0.05)，对减缓癫痫发作脑损伤有一定的神经保护作用。