低温-电场在神经干细胞移植中的优化作用及基于GAS5-miR142-MEK/ERK通路的分子机制研究

神经干细胞（NSCs）移植是颅脑创伤（TBI）后神经修复再生的重要潜在手段，目前优化移植的方案多聚焦于对细胞本身的分子修饰、联合药物应用等，效果尚不满意。基于我们前期物理因素干预神经细胞的系列研究发现及预实验结果，包括低温抑制炎症和促进神经突起生长，电场调控细胞运动方向和能力，我们提出：探索低温-电场干预优化TBI后NSCs移植及其分子机制。围绕此科学问题，1）通过体外细胞实验，研究低温-电场干预对人源性NSCs（hNSCs）存活、增殖、分化、运动特性的影响，并聚焦于GAS5-miR142-MEK/ERK通路探索内在机制；2）构建神经细胞损伤模型、大/小鼠控制性皮层损伤模型，研究低温-电场对hNSCs在TBI环境下的作用和机制；3) 明确最佳的移植前/后的低温-电场干预方案。本研究首次联合应用物理干预对hNSCs移植进行系统性优化，为低温-电场应用于hNSCs移植提供理论依据和优化新策略。

Neural stem cells (NSCs) transplantation is a very promising therapy for nerve repair and neural regeneration after traumatic brain injury (TBI). Currently, the majority of the optimizing schemes for transplantation are focusing on the molecular modification of NSCs or the application of medical reagents. However, the optimizing effects of these schemes are not satisfying. Based on our previous findings and pilot study on physical interventions on neural cells, we have found that hypothermia can inhibit inflammatory responses and promoted neurite growth, while electric field (EF) can regulate cell migration significantly. Herein, we propose to research the optimize effect of hypothermia and EF on the transplantation of NSCs and its molecular mechanisms. Focusing on this scientific problem, this project will: 1) concentrate on the effects of hypothermia and EF on human NSCs (hNSCs) viability, proliferation, differentiation and motility in vitro, and explore the underlying molecular mechanisms focusing on the GAS5-miR142-MEK/ERK signaling pathway; 2) explore the optimizing effects and mechanisms of hypothermia and EF on hNSCs under the TBI environment by using established injury models on neural cells and controlled cortical injury models on mice and rats; 3) identify the optimal hypothermia and EF intervention protocols before and after transplantation. This study is the first to jointly apply physical interventions to systematically optimize hNSCs transplantation, providing a theoretical basis for the application of hypothermia and EF in hNSCs transplantation, and a novel strategy in neural regeneration.