淫羊藿、黄芪、葛根有效组分复方通过诱导HAMP抑制APP淀粉样代谢通路改善阿尔茨海默病的机制研究

Brain iron overload in AD patients were related to the loss control of iron metabolism related proteins, but the exact mechanism is unclear. In our previous studies, we found thatEpimedium herb, astragalus root, pueraria effective component compound can alleviate the iron overload in AD models by increasing the expression of PFN1, and reduce the expression of Aβ. HAMP is an important iron regulating hormone, which can promote the degradation of PFN1. Aβ is the central factor of AD, which produced by the typical metal protein APP. Our preliminary experimental results showed thatEpimedium herb, astragalus root, pueraria effective component compound could up-regulate the expression of HAMP and down-regulate the expression of APP. Therefore, we hypothesized that Epimedium herb, astragalus root, pueraria effective component compound could induce HAMP to interfere with brain iron metabolism, inhibit the pathway of APP amyloidosis, and reduce the formation of Aβ, which may be an important mechanism for the development of AD. Therefore, this study HT22 with knockdown HAMP was used to establish the AD cell model in the cell level, and the AD transgenic mice with HAMP knockout mice hybrid F1 generation in animal level, research the intervention effect of HAMP on the key protein in the APP metabolic pathways, discusses theEpimedium herb, astragalus root, pueraria effective component compound intervention mechanism of brain iron homeostasis by inducing HAMP regulates the generation of Aβ to reveal the HAMP intervention brain iron metabolism involved in the pathogenesis of AD experimental basis, for the development of new offers the formulas for the treatment of AD.

AD患者脑铁水平异常升高与脑内铁代谢相关蛋白表达失控相关，但具体机制不明。我们前期研究发现淫羊藿、黄芪、葛根有效组分复方可上调PFN1表达缓解AD小鼠脑铁超载，同时下调Aβ表达。HAMP是重要的铁调节激素，可促进PFN1降解。Aβ是AD发病的中心环节，由典型金属蛋白APP经淀粉样代谢通路产生。预实验结果显示该复方可上调HAMP表达，下调APP表达。由此我们推测该复方通过诱导HAMP干预脑铁代谢，抑制APP淀粉样代谢通路减少Aβ生成可能是该复方缓解AD发生发展的重要机制。因此，本课题细胞水平以敲低HAMP的HT22建立AD细胞模型，动物水平以AD转基因小鼠与HAMP基因敲除小鼠杂交F1代为研究对象，研究HAMP对APP代谢通路中关键蛋白的干预作用，探讨该复方诱导HAMP干预脑铁代谢调控Aβ生成的机制，为揭示HAMP干预脑铁代谢参与AD发病机制奠定实验基础，为开发治疗AD的新方药提供思路。

阿尔茨海默病（Alzheimer’s Disease, AD）患者脑铁水平异常升高与脑内铁代谢相关蛋白表达失控相关，但具体机制不明。我们前期研究发现淫羊藿、黄芪、葛根有效组分复方可上调PFN1表达缓解AD小鼠脑铁超载，同时下调Aβ表达。hepcidin是重要的铁调节激素，可促进PFN1降解，该复方亦可上调hepcidin表达，下调APP表达。Aβ是AD发病的中心环节，由典型金属蛋白APP经淀粉样代谢通路产生。由此我们推测该复方通过诱导hepcidin干预脑铁代谢，抑制APP淀粉样代谢通路减少Aβ生成可能是该复方缓解AD发生发展的重要机制。因此本研究利用中药系统药理学数据库与分析平台(TCMSP)、BATMAN-TCM生物信息学分析工具收集淫羊藿、黄芪、葛根3味中药主要成分及潜在的作用靶点，并将其与AD相关基因进行映射，得出3味中药治疗AD的作用靶标，构建蛋白-蛋白相互作用网络并进行可视化。证实淫羊藿、黄芪、葛根均有直接治疗AD的作用，且效果与西药类似。进一步采用APPswe/PS1dE9双转基因AD模型小鼠及同Aβ25-35共孵育的小鼠海马神经元HT22神经细胞模型为评价载体，以铁调素hepcidin为切入点，采用神经分子生物学研究技术和手段，对APP水解关键酶及相互作用蛋白及mRNA进行了检测，阐明了铁调素hepcidin在调节APP代谢通路中发挥着重要作用，并明确了基于虚-痰-瘀-毒病机理论配伍的淫羊藿、黄芪、葛根有效组分复方对APPswe/PS1dE9双转基因AD模型小鼠及海马神经元的干预作用与调节APP代谢通路有关。