老年性痴呆脑微环境对海马移植NSCs的作用及针刺干预研究

神经干细胞（NSCs）移植为神经再生修复治疗带来巨大前景，但只有少量细胞能长期存活并发挥功能，最终影响痴呆治疗的效果。植入NSCs的命运很大程度取决于微环境，微环境密切调节着NSCs移植的进程。本课题在针刺治疗老年性痴呆的确切临床疗效和针刺调节内源性NSCs的实验基础上，运用"针刺调理三焦改善脑髓不充"的中医理论，将针刺与NSCs移植相结合，从针刺影响NSCs移植微环境角度，以SAMP8为研究对象，应用免疫印迹和实时荧光定量PCR技术观察针刺对微环境指标的影响，通过体内移植NSCs和建立海马脑片与NSCs共培养体外模型两方面，应用HE染色、免疫荧光、免疫组化、流式等技术观察针刺对移植NSCs存活、增殖、分化和形成突触网络的影响，分析针刺改善NSCs移植微环境和改善痴呆鼠认知障碍的内在联系，从微环境角度为探讨针刺调节NSCs移植的抗痴呆机制及针刺三焦与脑病的相关性等提供新思路和实验依据。

Neural stem cells (NSCs) transplantation has brought about a huge prospect for the nerve regeneration treatment. However, there is no large number of newborn neurons formations in the brain of host after NSCs transplantation, and ultimately affect the efficacy of transplantation for the treatment of dementia. The ultimate fate of implanted NSCs largely depended on the surrounding microenvironment, and the microenvironment closely regulated the process of NSCs transplantation. In this topic, basing on the exact clinical efficacy of acupuncture treatment for senile dementia and the experimental result of acupuncture regulation of endogenous NSCs, we apply "acupuncture regulating Sanjiao and improving brain insufficient" Chinese medicine theory, and combine acupuncture with NSCs transplantation. We are going to use SAMP8 from the view of acupuncture regulating NSCs transplantation microenvironment. We will apply the western blot and real-time quantitative PCR technique to observe the effects of acupuncture on NSCs microenvironment related indicators. And we will apply HE staining, immunofluorescence, immunohistochemistry and flow cytometry techniques to observe the effect of acupuncture on the survival, proliferation, differentiation and the formation of the synaptic network of transplanted NSCs from both NSCs transplantation in vivo and hippocampal slices with NSCs co-cultured in vitro. After that, the inherent connections between the improvement of the NSCs transplantation microenvironment and the improvement of the cognitive impairment of dementia mouse by acupuncture will be analyzed. This will provide new ideas and experimental evidence for the anti-dementia mechanism of acupuncture regulating NSCs transplantation and for the correlation of acupuncture Sanjiao and cerebral diseases.