Nrf2调控神经干细胞（NSCs）存活和分化发挥脑缺血性卒中后神经修复的作用和机制研究

Cerebral ischemic stroke is one of the most severe brain vascular diseases with high disability and mortality. In recent years, neural surgeons have made great progresses in acheiving early reflow by systematic thrombolysis and endovascular treatments; As to the existed neuronal injury, there is no helpful treatment. The transplantation of neural stem cells (NSCs) has showed great potential in the treatment of neuronal injury, which promotes neuron repair and function recovery. However, the mechanism is not completely clear. the low survival rate of the transplanted NSCs and the the microenvironment limits its effect. Nrf2 is a transcription factor of a variety of antioxidant enzymes, which have been reported to play an important role in the survival and differentiation of NSCs. After paper reviewing and preliminary studying, we aim to establish cerebral ischemic stroke model, and then use the methods of behavior, morphology, molecular biology, cellular immunology, to explore the relationship between Nrf2 and neuronal repair induced by endogenous and exogenous NSCs. Additionally, we will try to clarify the regulation effect of Nrf2 on the proliferation and differentiation of NSCs, and the mechanism in promoting nerve repair. Finally, we will explore the combined therapeutic strategy of Nrf2 activation and NSCs transplantation after cerebral ischemic stroke. Our study will provide new experimental and theoretical basis for the clinical treatments of stroke.

脑缺血性卒中致残、致死率高，社会危害性大，近年来，其临床治疗不断进展，已实现超早期血流再通。但对于既成的神经损伤，临床往往束手无策。神经干细胞（NSCs）移植在治疗神经损伤方面具有巨大潜力，可以促进神经修复，改善功能障碍。但是，作用机制不明，移植NSCs存活率不高，分化受微环境影响，导致其作用受限。Nrf2是多种抗氧化酶的转录因子，研究提示其在NSCs存活、分化中发挥重要作用。本课题组结合文献和前期研究，拟通过构建脑缺血性卒中模型，综合运用行为学、形态学、分子生物学、细胞免疫学等技术，研究Nrf2与卒中后内、外源性NSCs发挥神经修复作用的关系，明确Nrf2对NSCs增殖分化的调控作用，以及其促进神经修复机制，并继续探索激动Nrf2联合NSCs移植治疗脑缺血性卒中后神经损伤策略，为临床治疗脑卒中提供新的实验依据和理论基础。

缺血性脑卒中致残、致死率高，社会危害性大，近年来，其临床治疗不断进展，已实现超早期血流再通。但对于既成的神经损伤，临床往往束手无策。神经干细胞（NSCs）移植在治疗神经损伤方面具有巨大潜力，可以促进神经修复，改善功能障碍。但是，作用机制不明，移植NSCs存活率不高，分化受微环境影响，导致其作用受限。Nrf2是多种抗氧化酶的转录因子，其在NSCs存活、增殖、分化中发挥重要作用。本课题主要研究了3个方面的内容：（1）Nrf2及其下游信号通路对NSCs增殖及分化能力的影响，以及对MCAO小鼠神经功能、氧化应激、炎症水平的影响；（2）侧脑室注射NSCs对MCAO小鼠神经功能的保护作用及其机制；（3）侧脑室注射天然Nrf2激动剂紫檀芪联合NSCs注射对MCAO小鼠神经功能的保护作用及其机制。在研究内容的基础上，本课题分离培养了小鼠NSCs，通过AAV载体过表达或干扰Nrf2，以MCAO模型为基础，立体定向侧脑室注射药物、NSCs及AAV载体，通过神经功能评分、形态学染色、WB、ELISA、免疫荧光染色等方法，明确了Nrf2信号通路，以及外源性NSCs对MCAO后神经损伤的修复作用；阐述了Nrf2信号通路对NSCs增殖、分化的调控作用，以及MCAO后Nrf2在海马区的表达变化，揭示了Nrf2信号通路可能为脑缺血性卒中后神经修复的重要机制；探索了天然Nrf2激动剂紫檀芪联合NSCs侧脑室注射治疗脑缺血性卒中后神经损伤的策略，并阐述了其调节神经炎症、减轻氧化应激以及促进移植物NSCs增殖分化的潜在机制，为临床治疗脑缺血性卒中提供新的方向和理论依据。