在骨髓间充质干细胞过表达CX3CL1 与Wnt3a基因调节小胶质细胞活性促进神经元再生

项目名称： 在骨髓间充质干细胞过表达CX3CL1 与Wnt3a基因调节小胶质细胞活性促进神经元再生

项目编号： No.81471236

项目类型： 面上项目

立项/批准年度： 2015

项目学科： 医药、卫生

项目作者： 陆大祥

作者单位： 暨南大学

项目金额： 70万元

中文摘要： 研究表明，小胶质细胞的过度激活是阿尔茨海默病（AD）的重要发病机制之一，而趋化因子（C-X3-C基元）配体1(CX3CL1)的信号衰减与小胶质细胞的活化和海马神经元的损伤密切相关。我们前期研究已证实，上调Wnt3a基因可增加趋化因子受体1 （CX3CR1）的表达，通过调控小胶质细胞活性可减轻神经炎症有助于神经再生。因此，本项目拟在骨髓间充质干细胞（BMSCs）以慢病毒为载体高表达CX3CL1 与Wnt3a基因，与小胶质细胞共培养，探讨BMSCs对Aβ诱导的小胶质细胞活化调节作用；采用荧光共聚焦技术，探讨Aβ诱导下高表达CX3CL1与Wnt3a基因的BMSCs对海马神经祖细胞增殖与分化的调控作用；并移植入AD模型转基因小鼠海马，检测神经再生以及小鼠记忆和行为学改变。通过干细胞传递治疗性蛋白方法，将抑制神经炎症与促进神经再生作用相结合防治AD，拟为AD的防治提供重要的理论基础和新的治疗策略。

中文关键词： 趋化因子配体1；小胶质细胞；神经炎症；神经再生；阿尔茨海默病

英文摘要： Alzheimer's disease (AD) patients suffer from irreversible injuries in cognition and memory because of the aggregation of abnormal proteins and over-activated microglia. Recent evidence support that the decrease in CX3CL1/CX3CR1 signaling pathway is associated with over-activated microglia and hippocampal neurons injures. Our recent studies also showed that regulation of over-activated microglia facilitate the survival of neurons. Therefore, it is meaningful to regulate CX3CL1/CX3CR1 signaling pathway in neuroinflammation and neurogenesis in AD. In this proposal, using luciferase reporter system, we attempt to screen the signal proteins that upregulate CX3CR1 gene expression. We will use lentiviral- trangentic vector system to overexpress CX3CL1 and Wnt3a in BMSCs, and then explore the regulation of Aβ protein-induced pro-inflammatory factor TNF- α, IL-1 β and IL-6 by genetically modified BMSCs. In vitro, we will analyze the expression of neural progenitor cell markers Sox2, Nestin and DCX, and proliferation cell markers Ki67 and EdU incorporation by confocal, and then evaluate the effect of genetically modified BMSCs on the regulation of differentiation and proliferation in neural progenitor cells. In vivo, we will transplant genetically modified BMSCs into hippocampus in APP/PS1 transgenetic mice to study neurogenesis in hippocampus. Furthermore, the changes in memory and behavior after transplantation will be studied. This proposal using stem cells to deliver therapeutic proteins, and combining the inhibition of neuroinflammatory injuries and induction of neurogenesis will provide a new therapeutic strategy for AD.

英文关键词： CX3CL1;microglia;Neuroinflammation;Neuroregeneration;Alzheimer's disease