从神经干细胞分化研究黄芪多糖调控CPZ脱髓鞘小鼠髓鞘再生的效应机制

多发性硬化是中枢神经系统炎性脱髓鞘疾病，临床缺乏有效治疗。少突胶质细胞OL死亡是引发脱髓鞘的直接因素，调控神经干细胞NSC向OL分化是促进髓鞘再生、治疗脱髓鞘疾病的关键。本项目在前期研究基础上，拟建立CPZ诱导的脱髓鞘小鼠模型，从体重变化、运动功能、平衡功能等行为学方面，探讨黄芪多糖对CPZ脱髓鞘小鼠的治疗作用；采用形态学和免疫组织化学方法，探讨黄芪多糖对CPZ脱髓鞘小鼠髓鞘再生的影响；建立NSC体外培养系统，从体内和体外实验，探讨黄芪多糖调控NSC向OL分化的效应；运用分子生物学方法，进一步探讨黄芪多糖对Shh信号通路关键分子的调控，藉以阐明黄芪多糖促进髓鞘再生、调控NSC分化的效应机制，揭示黄芪多糖治疗脱髓鞘疾病的新靶点。本项目是前期工作的延续和深入，属跨学科的前沿课题，具有鲜明创新性，至今尚无报道，为临床疑难疾病的治疗和新药研发提供了实验依据，对于促进中医药学发展具有重要的科学意义。

Multiple sclerosis (MS) is an inflammatory demyelinating disease in the central nervous system. There is not any effective therapeutic approach in treating the disease up to now. The death of oligodendrocyte (OL) is the direct factor for initiation of demyelination. Therefore regulating differentiation of neural stem cells (NSC) into OL is a crucial way to promote the remyelination and to treat the demyelinating diseases effectively. In our previous studies it was found that astragalus polysaccharide (APS) could significantly suppress the disease of demyelinating animal model of MS, and could effectively reduce the inflammatory demyelination. Based on the previous studies, this project aims to investigate the effects and the mechanisms of APS in promoting remyelination and regulating differentiation of NSC into OL. In this project, CPZ induced demyelinating model in mice will be established. The therapeutic effects of APS on the CPZ-demyelinating mice will be observed from the behavioral aspects, such as the weight changes, motor function, and balance function of the mice model. Meanwhile, the morphological and immunohistochemical methods will be used to observe the promoting effects of APS on remyelination in the CPZ-demyelinating mice. Furthermore, the culture system of neural stem cells in vitro will be set up so that the experiments both in vivo and in vitro will be launched out. The expression of Nestin, NeuN, GFAP, and MBP, the molecular markers of neural stem cells, neurons, astrocytes, and oligodendrocytes, will be detected respectively to determine regulatory effects of APS on differentiation of neural stem cells. The effects of APS on the Shh signaling pathway will also be observed by assaying the expression of the key molecules using molecular biological techniques and immunocytochemistry methods. It will demonstrate the effects and the molecular mechanisms of APS in treating the CPZ-demyelinating mice, promoting remyelination, and regulating the differentiation of neural stem cells to oligodendrocytes in the CPZ-demyelinating mice. This project will reveal the new therapeutic target of APS in treating autoimmune inflammatory demyelinating diseases in the central nervous system, which will provide the important scientific evidence for the research and development of new drugs in clinic. It will be an interdisciplinary, innovative, frontier research project, which is of great significance to promote the modernization of traditional Chinese medicine.