健脾益智法调控局灶性脑缺血大鼠脑神经干细胞增殖分化诱导神经再生的分子机制研究

In the theory of TCM，five visceras have these functions of production, growth，gasification and preservation, and among them the function of gasification belongs to spleen , scilicet, spleen governing movement and transformation.The spleen is viewed as the source for the production and transformation of qi , blood and the body fluid, warming nourishing, promoting and stimulate all the Organs including the brain, and is also closely related to the channel and collateral, configuration and constitution of the body and limbs and skeleton.So the proliferation and differentiation of neural stem cells and neural regeneration should be in the range of the theory that spleen controls the transformation and growth.In Our previous research to treat Alzheimer's disease by the Jianpi Yizhi Method,we concluded that it is an important method to protect damaged neurons through the modulating of the proliferation and differentiation of the neural glia cells,so whether the theory of spleen controlling the transformation and growth can be used in the treatment of IS,exploring the mechanism on neural regeneration seems much important. Under the guidance of the spleen controling the transformation and growth,we make the focal cerebral ischemic rats to be research objects,observe the expression of the exogeous cytokine and endogenous gene to explore the mechanism of Jianpi Yizhi Method modulating the proliferation and differentiation of neural stem cells,in order to reveal the relationship between the spleen and the cerebral neural regeneration,find the micro-mechanism of spleen controlling the transformation and growth,provide new ideas and methods for the prevention and treatment of IS.

中医认为：五脏具有生长化收藏的作用，其中"化"属于脾，即脾主化，能够化生人体精气血津液等精微物质，推动、激发并荣养包括脑在内的五脏六腑、形体官窍，以及全身经络组织等，所以脾的这种"化生"作用对于脑NSCs的增殖、分化同样也具有重要的调节和促进作用。结合课题组前期开展的健脾益智法治疗多发性梗塞性脑病的研究中，明确了健脾益智法调控神经胶质细胞的活化增殖是保护受损脑神经元的重要途径之一，是治疗缺血性脑病的有效方法，本项目拟在中医"脾主化生"理论指导下，以"脑NSCs的增殖分化是脑神经再生的重要途径"为切入点，采用现代生物学技术，观测健脾益智法对缺血性模型大鼠行为学、脑NSCs增殖及神经元新生的影响，并进一步从外源性细胞因子和内源性基因的表达入手，探讨其分子调控机制，籍以阐明"脾主化生"理论的科学内涵，揭示脾的"化生"作用与脑神经再生的关系，为临床防治缺血性脑卒中提供新的思路和方法。

中医认为：五脏具有生长化收藏的作用，其中"化"属于脾，即脾主化，能够化生人体精气血津液等精微物质，推动、激发并荣养包括脑在内的五脏六腑、形体官窍，以及全身经络组织等，所以脾的这种"化生"作用对于脑NSCs的增殖、分化同样也具有重要的调节和促进作用。本项目在中医"脾主化生"理论指导下，以"脑NSCs的增殖分化是脑神经再生的重要途径"为切入点，观测健脾益智法对缺血性模型大鼠行为学、脑NSCs增殖及神经元新生的影响，并进一步从外源性细胞因子和内源性基因的表达入手，探讨其分子调控机制，发现：（1）健脾益智胶囊能够促进MCAO大鼠学习记忆能力的恢复。（2）健脾益智胶囊能够促进脑NSCs的增殖并向神经元分化。（3）健脾益智胶囊能够提高Notch1、Jagged1、β-catenin蛋白及基因的表达。（4）健脾益智胶囊可促进NT-3、BDNF、EGF、bFGF的表达。（5）健脾益智胶囊可促进NT3、GAP-43、SYP-P38、PKA的表达。（6）健脾益智胶囊可抑制IL-6、NF-KB的表达。