基于microRNA负调控Notch和Wnt双信号转导通路探讨针刺改善老年性痴呆海马微环境调节移植NSCs生物学功能的作用机制

神经干细胞（NSCs）移植为神经修复再生带来巨大前景。课题组前期发现针刺对痴呆小鼠认知障碍的改善与调节外源性NSCs在海马微环境的生物学功能有关。Notch和Wnt信号是调节NSCs生物学功能的重要通路且存在交互作用，而microRNA通过靶向上述信号发挥负调控作用。本课题在针刺治疗老年性痴呆（AD）的确切临床疗效基础上，运用“针刺调理三焦气化改善脑髓不充”中医理论，取穴膻中、中脘、气海，血海和足三里，通过外源性NSCs移植和建立海马脑片与NSCs共培养体外模型两方面，应用免疫荧光、免疫印迹，实时定量PCR、流式等技术观察针刺对SAMP8海马微环境miRNA-124及其靶向双信号关键分子的影响，分析针刺调节miRNA靶向信号通路而改善海马微环境与调控外源性NSCs发挥生物学功能的内在联系，从表观遗传学角度为探讨针刺改善微环境调节NSCs的抗痴呆机制和NSCs移植早期针刺临床干预提供依据。

The transplantation of neural stem cells (NSCs) has brought about a great prospect for nerve repair and regeneration. Our previous studies have shown that acupuncture could improve the cognitive impairment of SAMP8 mice, which might be related to the biological function regulation of exogenous NSCs in the hippocampus microenvironment. Notch and Wnt signals are important pathways to regulate the biological function of NSCs, and there is interaction between them. While some microRNA negatively regulates the biological behavior of NSCs by targeting the above signaling pathways genes. In this study, basing on the exact clinical efficacy on senile dementia by Sanjiao acupuncture, we use "acupuncture regulating Sanjiao Qihua and improving brain insufficient" theory of Traditional Chinese Medicine, and select "Danzhong", "Zhongwan", "Qihai", "Xuehai" and "Zusanli" acupoints. We apply exogenous NSCs transplantation in vivo and establish co-cultured model of hippocampal slices and NSCs in vitro. We use immunofluorescence, Western blot, Real-time PCR and flow cytometry to evaluate the effects of acupuncture on the expression of miRNA and its targeted key molecules of the dual signal pathways in SAMP8 hippocampus microenvironment, in order to analyze the relationship of acupuncture regulating miRNA targeting signal pathways, improving hippocampus microenvironment and regulating exogenous NSCs to play biological functions. Thus, this study can provide the evidence and the basis for the anti-dementia mechanism of acupuncture to improve the NSCs microenvironment and the clinical intervention of acupuncture in the early stage of NSCs transplantation, from the perspective of epigenetics.