基于Notch信号通路抑制的脾虚证食欲中枢机制研究

It has been increasingly recognized that gastrointestinal system functions that included substance and energy metabolism, dysbacteriosis and immune disorders played pivotal factors in the generation of the syndrome of spleen deficiency. However, the mechanism underlying the syndrome of spleen deficiency has not been elucidated from the perspective of the dysfunction in the feeding center. As reported previously, the appetite regulation network of arcuate nucleus (Arc) in the hypothalamus played an important role on the syndrome of stagnation of liver qi and spleen deficiency, during which Notch signal pathway involoved in regulating the feeding and energy metabolism disorders. Thus, the study proposes the hypothesis that the syndrome of spleen deficiency-induced the feeding and energy metabolism disorders were regulated by the appetite regulation network of hypothalamic Arc, where the inhibition or activation of Notch signal pathway may be one of the underlying mechanisms. The rat molde of syndrome of spleen deficiency was established in accordarnce with the TCM animal model of complex causes of disease. The Notch signal pathway blocker was used as a vital experimental tool. The Sijunzi decoction and mosapride were separately administrated as the intervention and positive control treatment. In the experimental result, the changes in the morphology of hypothalamic Arc, the level of related appetite regulation factors, the macro representation, the level of related feeding factors in peripheral tissues and the related energy metabolism enzymes were determined. In summary, the purpose of the study aims at revealing that the dysfunction in hypothalamic Arc, a key feeding center, may be one of the mechanism underlying the generation of the syndrome of stagnation of liver qi and spleen deficiency. Nevertheless, importantly, the inhibition or activation of Notch signal pathway may be a vital regulating point in the process.

对脾气虚证的研究，焦点多集中在胃肠道物质代谢和能量代谢，以及胃肠道菌群、肠道免疫等方面，较少涉及中枢神经系统。结合前期研究发现：肝郁脾虚证与下丘脑弓状核（Arc）“食欲调节网络”密切相关，Notch信号通路参与了摄食和能量代谢紊乱的调节，本项目提出“脾虚证的摄食和能量代谢紊乱是受下丘脑Arc食欲调节网络调控，Notch信号通路的抑制/激活是其机制之一”的假说。选取复合病因因素建立脾虚证大鼠模型，Notch信号通路阻断作为研究手段，以中药四君子汤进行干预，西药莫沙必利作为阳性对照，从下丘脑Arc形态学的改变，食欲调节因子水平的变化，宏观表征的改变及外周摄食相关因子和能量代谢相关酶的变化，揭示下丘脑Arc食欲中枢功能的紊乱可能是脾虚证形成的中枢机制之一，而Notch信号通路的抑制/激活可能是其中的关键分子事件。

1.研究背景.目前对脾虚证的研究，焦点多集中在胃肠道物质代谢和能量代谢，以及胃肠道菌群、肠道免疫等方面，较少涉及中枢神经系统。结合前期研究发现：肝郁脾虚证与下丘脑弓状核（Arc）“食欲调节网络”密切相关，Notch信号通路参与了摄食和能量代谢紊乱的调节，本项目提出“脾虚证的摄食和能量代谢紊乱是受下丘脑Arc食欲调节网络调控，Notch信号通路的抑制/激活是其机制之一”的假说。.2.主要研究内容.采用复合病因因素建立脾虚证大鼠模型，Notch信号通路阻断作为研究手段，以中药四君子汤进行干预，西药莫沙必利作为阳性对照，观察下丘脑Arc形态学的改变、食欲调节因子水平的变化，观察宏观表征的改变及外周摄食相关因子、能量代谢相关酶和B族维生素的变化。.3.重要结果.（1）饮食失节+疲劳游泳的造模方法可以使大鼠体重、摄食、体温、小肠吸收功能及自主活动水平均显著下降，成功复制了脾虚证模型；四君子汤和莫沙必利能够改善以上脾虚证症状；Notch信号通路抑制后，动物的活动能力水平恢复显著。.（2）脾虚证会引起下丘脑Arc Notch信号通路的激活，进而引起促进食欲的NPY表达降低，抑制食欲的POMC表达升高；当Notch信号通路被阻断后，这两种食欲太的异常表达均被逆转。.（3）脾虚证会引起外周抑制食欲的Leptin表达升高，激发食欲的Ghrelin表达降低，代谢酶及B族维生素水平也相应降低；当Notch信号通路被阻断后，这些异常表达被逆转。.4.科学意义.揭示了下丘脑Arc食欲中枢功能的紊乱可能是脾虚证形成的中枢机制之一，而Notch信号通路的抑制/激活可能是其中的关键分子事件，对临床运用四君子汤治疗脾虚提供科学依据具有重要意义。.