鹿茸多肽诱导间充质干细胞定向分化为神经元的机理研究

Mesenchymal stem cells (MSCs) are capable of self-renewal and multilineage differentiation. With these considerations in mind, stem-cell-based therapies for senile dementia could provide suitable therapeutic approaches. Presently, molecular mechanisms underlying neurons specification are still poorly understood, which will impede experimental research and clinical application of MSCs，so it is necessary to understand the mechanism of neural differentiation of MSCs. On the basis of preliminary studies, the project aims to the extracts of pilose antler polypeptides for neural differentiation of MSCs in vitro. we will study biologic activities of pilose antler polypeptides, for example, promoting MSCs' proliferation and neural differentiation assisting differentiation technology of stem cells, research neural features of differentiated MSCs assisting cell chemical, western blot and molecular biology methods, studying neural differentiation of mesenchymal stem cell combined flow cytometric, study electrophysiological characteristics of differentiated MSCs by patch-clamp technique, study signaling pathway of neural differentiation of MSCs, treated with active components of velvet antler polypeptides, assisting inhibitor and excited agent of mTOR. Results will prove that our viewpoint of mTOR and its downstream molecules are involved in neural differentiation of MSCs. Our study will facilitate development of new drug of velvet antler and clinical application of MSCs.

间充质干细胞可诱导定向分化为神经元，体内移植可修复老年痴呆患者受损脑组织、重建细胞环路和细胞功能，用于治疗老年性痴呆。阐明间充质干细胞定向分化为神经元的机制有利于推动间充质干细胞的临床应用。 我们前期研究发现，鹿茸多肽具有诱导间充质干细胞定向分化为神经元的作用。通过体外间充质干细胞定向分化条件平台技术，研究鹿茸多肽诱导间充质干细胞定向分化为神经元的作用；采用细胞化学、基因转染、免疫印迹等分子生物学技术研究分化细胞（即已经分化为神经元的间充质干细胞，下同）的神经元样功能；通过膜片钳技术研究分化细胞的电生理特征；通过流式细胞术研究间充质干细胞的分化率；结合mTOR的抑制剂和激动剂，研究鹿茸多肽诱导间充质干细胞定向分化为神经元的信号通路作用靶点，进一步证明"mTOR及其下游分子在间充质干细胞定向分化为神经元中发挥重要作用"的理论，为间充质干细胞的临床应用以及鹿茸多肽新药的研究开发奠定基础。

间充质干细胞可诱导定向分化为神经元，体内移植可修复老年痴呆患者受损脑组织、重建细胞环路和细胞功能，用于治疗老年性痴呆。阐明间充质干细胞定向分化为神经元的机制有利于推动间充质干细胞的临床应用。 .我们研究发现，鹿茸多肽具有诱导间充质干细胞定向分化为神经元的作用。通过体外间充质干细胞定向分化条件平台技术，研究鹿茸多肽诱导间充质干细胞定向分化为神经元的作用；采用细胞化学、基因转染、免疫印迹等分子生物学技术研究分化细胞的神经元样功能，分化细胞表达表达神经元细胞特异性的蛋白，分泌NGF和GDNF等生长因子，表达表达神经元细胞特异性的蛋白 β 微管蛋白（βTubulin-III）、神经元中间丝蛋白(NF)、神经元特异性烯醇化酶(neuron specific enolase, NSE)；通过研究鹿茸多肽诱导间充质干细胞定向分化为神经元的信号通路作用靶点，进一步证明mTOR 及其下游分子在间充质干细胞定向分化为神经元中发挥重要作用”的理论，为间充质干细胞的临床应用以及鹿茸多肽新药的研究开发奠定基础。