基于神经干细胞巢内notch通路与caveolin-1串话研究"虚、瘀、郁"状态下中风并发抑郁促神经再生及中药干预

Neural stem cells in the nest is through a complex signal transduction regulating the proliferation of neural stem cells (NSCs) microenvironment, caveolin-1 is the NSCs signal transmission hub, but the mechanism is unknown. Our previous studies have shown that stroke in the "deficiency, blood stasis, yu" easily complicated with depression, on the basis of further neurologic deficits. Caveolin-1 can regulate the Notch pathway, Hes Notch-1 of expression, alleviate cerebral ischemia injury, promote neural functional recovery and nerve regeneration, but its molecular mechanism is unclear. Notch is the key to a regulation of cell proliferation, differentiation, conservative pathway, regulation can be caveolin through receptor-1, crosstalk between the two. The Notch pathway and the crosstalk of caveolin-1 is the regulating mechanism of nerve regeneration after cerebral ischemia? Therefore, this topic proposed using RNA interference technology, molecular functional imaging techniques and 3D nano cell culture, specificity of antagonism and immune coprecipitation method in neural stem cells nest inside Notch with caveolin-1 crosstalk effects on nerve regeneration and refreshing XieYu party to promote the mechanism of nerve regeneration, reveal the mechanisms regulating nerve regeneration after cerebral ischemia and new targets.

神经干细胞巢是通过复杂的信号传导调控神经干细胞（NSCs）增殖的微环境，caveolin-1是NSCs信号传递枢纽，但作用机制不详。我们前期研究显示，中风病在"虚、瘀、郁"的基础上易并发抑郁，进一步加重神经功能缺损。caveolin-1能调控Notch通路中Notch-1、Hes-1的表达，减轻脑缺血损伤、促进神经功能恢复和神经再生，但其分子机制尚不明确。Notch是一条调控细胞增殖、分化的关键、保守通路，能通过受体被caveolin-1调控，二者之间存在串话。那么Notch通路与caveolin-1的串话是其调控脑缺血后神经再生的机制吗？故本课题拟运用RNA干扰技术、分子功能影像技术、3D纳米细胞培养、特异性拮抗、免疫共沉淀等方法探讨在神经干细胞巢内Notch与caveolin-1串话对神经再生的影响及醒脑解郁方促神经再生的机理，揭示脑缺血后神经再生的调节机制和新的作用靶点。

卒中后抑郁症（Post-stroke depression,PSD）是脑卒中最常见的并发症之一，严重影响卒中的康复。神经干细胞巢是通过复杂的信号传导调控神经干细胞（NSCs）增殖的微环境，主要包括附近的支持细胞、胞外基质和微血管网。caveolin-1是NSCs信号传递枢纽，调控Notch-1、Hes-1的表达，减轻脑缺血损伤、促进神经功能恢复和神经再生，但其分子机制尚不明确。我们前期研究显示，中风病在“虚、瘀、郁”的基础上易并发抑郁，进一步加重神经功能缺损。本项目采用大脑中动脉栓塞法联合慢性不可预见刺激制备PSD大鼠模型，行为学检测评估抑郁状态，PCR法检测大鼠海马Notch和Hes mRNA表达水平；Western blot法检测大鼠海马Notch和Hes蛋白水平，膜片钳技术检测大鼠海马Notch信号通路功能状态；此外，检测醒脑解郁方含药血清对神经干细胞迁移的影响。结果显示：醒脑解郁方能改善PSD大鼠行为学，增强脑内caveolin-1表达，激活海马组织Notch/Hes信号通路，促进内源性神经干细胞移行与神经再生。本研究揭示了神经干细胞巢内Notch与caveolin-1串话对神经再生的影响及醒脑解郁方促神经再生的机理，同时为PSD的治疗提供了新的作用靶点。