基于“脾主运化”原理的四君子汤调控肠-脑转运和脑能量代谢抗阿尔茨海默病的作用研究

Alzheimer's disease (AD) is seriously endangering the physical and mental health of the elderly, and is bringing a heavy burden to the society and the family. The pathogenesis of AD is complex, it might involve in β-protein (Aβ), Tau protein, material transport and metabolic disorder, chronic inflammation stimulation, even involve in disorder of neurotransmitter and brain energy metabolism. The frequent failure in AD drugs research and development basing on single target which indicated that research on AD drugs should base on multi-pathways, multi-targets. ‘Spleen governing transportation and transformation’ in Chinese medicine contains material transport and metabolism. Spleen deficiency will result in material transport and energy metabolism disorder. Literatures showed that invigorating spleen prescriptions have effect on AD. Are these related to the regulation of the transport of neurotransmitter and brain energy metabolism? It is still unclear. .Basing on our previous studies and combining ‘the spleen governing transport and transformation’ and the possible pathogenesis of AD, we proposed the hypothesis of ‘the function of spleen governing transportation and transformation’ containing the regulation of the transport of neurotransmitter in gut-brain axis and brain energy metabolism’. Sijunzi decoction, a classical prescription for replenishing Qi and invigorating the spleen, will be employed as a model. The transport of neurotransmitter associated with the Alzheimer's disease in gut-brain axis and brain energy metabolism will be further investigated using animal model, cell culture and microdialysis, Western Blot, ELISA, etc. The research will explain the scientific connotation of ‘spleen governing transportation and transformation’ from a new aspect, and provide a scientific basis for treating AD disease from spleen, provide a new idea for breakthrough AD drug research and development dilemma.

阿尔茨海默病（AD）严重危害老年人身心健康，给社会和家庭带来沉重负担。AD的病因复杂，不仅与β淀粉样蛋白、Tau蛋白、代谢转运紊乱、慢性炎症等有关，还与神经递质和脑能量代谢异常有关。针对单一靶点AD新药研发频频失败说明AD药物研发应立足于多靶点、多途径。中医“脾主运化”包含调控物质转运和代谢两方面，脾虚会引起物质转运和能量代谢异常。文献表明，健脾中药复方具有抗AD作用，这是否与其调控神经递质转运和脑能量代谢有关？尚不清楚。本项目在前期基础上，结合中医“脾主运化”及AD发病机制，提出“脾主运化与调控递质肠-脑转运和脑能量代谢关联”假说，以中药健脾基础方四君子汤为模型药，采用动物和细胞模型，运用微透析、Western Blot、ELISA等方法，重点研究四君子汤调控神经递质在肠-脑轴转运及脑能量代谢的作用，旨在揭示“脾主运化”的新内涵，为AD“从脾论治”提供依据，为AD药物研发提供新思路。

阿尔茨海默病（AD）严重危害老年人身心健康，给社会和家庭带来沉重负担。AD病因复杂，针对单一靶点AD新药临床研究频频失败。中医认为，“脾主运化”与体内物质转运和能量代谢相关。文献表明，健脾中药具有抗AD作用，但其机制尚不清楚。本项目提出“脾主运化与调控递质肠-脑转运和脑能量代谢关联”假说，立足AD发生与糖脂、能量代谢以及神经递质、脑肠肽关系，挖掘分析了脾与AD的关联性，并以健脾基础方——四君子汤为例，采用网络药理学、整体动物和细胞模型，从调控体内神经递质、脑肠肽、肠道菌群及血脑屏障上Aβ和葡萄糖转运蛋白等多角度，阐释了四君子汤抗AD的作用及机制，从全新角度揭示了“脾主运化”科学内涵，为“从脾论治”治疗AD提供科学依据，为破解AD新药研发困境提供新思路。研究结果如下：.1.四君子汤具有抗AD作用：可改善模型AD鼠学习记忆，减少海马神经元细胞损伤，降低血清和海马中Aβ含量，修复或对抗Aβ造成的细胞损伤。.2.四君子汤抗AD作用机制.（1）调控体内神经递质、脑肠肽和肠道菌群：能提高AD鼠血清和海马组织中乙酰胆碱、γ-氨基丁酸、5羟色胺含量，降低血清和海马组织谷氨酸含量，但对多巴胺无显著性影响；能提高AD鼠海马和结肠中胆囊收缩素、血管活性肠肽、Ghrelin等脑肠肽含量，降低AD鼠肠道中拟杆菌/类杆菌比例。.(2)调控血脑屏障上Aβ转运蛋白：对正常血脑屏障上Aβ转运蛋白（LRP1和RAGE）无明显影响，但对Aβ损伤的血脑屏障上转运蛋白有调控作用，可降低Aβ损伤的血脑屏障上RAGE表达，提高LRP1表达。.(3)调控胰岛素通路相关蛋白和葡萄糖转运蛋白：能控胰岛素水平关键通路PI3K/Akt，上调AD鼠海马p-PI3K/PI3K、p-Akt/Akt比值，增加PI3KmRNA、AKTmRNA及IRS-2 mRNA表达，提高AD鼠海马和皮质IRS-2平均光密度；对正常血脑屏障上葡萄糖转运蛋白（GLUT1和GLUT3）无明显影响，但可提高Aβ损伤的血脑屏障上GLUT1和GLUT3蛋白表达。