基于“毒损脑络”病机从Aβ/RAGE通路探讨黄连解毒汤协同抗AD物质基础及分子机制

The pathway of Aβ and the receptors for advanced glycation end products (RAGE), was an important toxic mechanism, and Aβ combining with RAGE can upregulate oxidative stress and inflammatory response. We have found that the generation and accumulation of Aβ were attenuated by use of berberine in Huanglian Jiedu Decoction. The Tau Protein hyperphosphorylation and the oxidative stress were attenuated in embryo rat neuron by use fo the flavonoids in Huanglian Jiedu Decoction, furthermore, the down-regulated activity of RAGE was confirmed by use of the flavonoids in the virtual screening and the results of their biological activity. So we hypothesized that “RAGE-Aβ signaling pathway may be involved in Huanglian Jiedu Decoction preventing the neuron from apoptosis in the treatment of AD”.Further, the material basis and molecular mechanism of Huanglian Jiedu Decoction based on the pathogenesis of RAGE-Aβ pathway in the treatment of AD were explored by use of IP and WB technology. The following cell signaling pathway are involved in the active components of Huanglian Jiedu Decoction preventing AD: Aβ/rage-Akt-GSK3β-p-Tau and Aβ/rage-ROS-cytoC-Apaf-caspase9,3 were further analyzed .The active components were screened and the mechanism was explored in order to systematically clarify the meaning of cerebral collaterals injured by toxin, to further reassemble the effective components based on RAGE-Aβ signaling pathway and achieve high efficiency and low toxicity in the treatment of AD.

Aβ与RAGE结合激活氧化应激、炎症等通路产毒而损伤脑络是AD致病关键。我们已发现黄连解毒汤君药黄连中小檗碱类能阻止Aβ生成、聚集；臣药黄芩中黄酮类抑制神经元细胞tau过磷酸化、氧化应激、保护线粒体，进而虚拟筛选和活性测试证实其能抑制RAGE活性，因此提出“黄连解毒汤成分通过调控RAGE-Aβ途径抑制神经元凋亡协同抗AD”假说。本项目拟进一步以Aβ损伤细胞及动物为模型，用IP、WB等技术，基于Aβ/RAGE筛选黄连解毒汤抗AD药效物质，探索机制及构效关系，重点关注其调控Aβ/rage-Akt-GSK3β-p-Tau、Aβ/rage-ROS-cytoC-Apaf-caspase9，3通路抗神经元凋亡机制。旨在明确该方干预RAGE-Aβ途径抗AD药效成分及机制，系统阐明君药祛毒之功、臣药减毒之效，揭示基于毒损脑络抗AD内涵，为筛选抗RAGE-Aβ通路药效成分并组方达到高效低毒抗AD提供科学依据

在本项目中，我们建立了斑马鱼活性初筛的模型，开展了药物有效成份五行属性及对AD治疗作用的探讨，确证了RAGE在线粒体途径中的调控作用及机制。以Aβ损伤细胞及动物为模型，在研究中，我们用IP、WB等技术，基于Aβ/RAGE筛选黄连解毒汤抗AD药效物质，重点研究了其调控Aβ/rage-Akt-GSK3β-p-Tau、Aβ/rage-ROS-cytoC-Apaf-caspase9，3通路抗神经元凋亡机制，明确该方干预RAGE-Aβ途径抗AD药效成分及机制，系统阐明君药祛毒之功、臣药减毒之效，揭示基于毒损脑络抗AD内涵，为筛选抗RAGE-Aβ通路药效成分并组方达到高效低毒抗AD提供科学依据。研究已证实，RAGE与Aβ相互作用，通过激活细胞内不同的信号通路，对阿尔茨海默病的发生发展发挥重要作用。