基于网络药理学与虚拟识别技术的黄芩三七协同治疗AD的靶向特性研究

It has been well recognized that Chinese herb components exert whole modulations which are additions of minor effects of the components. However the mechanism remains unclear. It can significantly increase the screen efficiency and find the most possible drug targets through analyzing the drug-related in vivo protein data with the viewpoints and techniques of network pharmacology; moreover, the drug-combinning proteins can be identified with the virtual identification through the reverse docking automatically search technique; thus the intersect set of the two above and the interaction network may be the integrated result of interactions of multiple primary and secondary targets.. This study is based on the recognized pharmacodynamical data that inhibited the synergic effects of baicalin and Panax Notoginsenosidum on senile dementia; and plans to build the drug-related network of differentially expressed proteins and the structure network of virtual identification through network pharmacological and virtul identification reverse docking techniques, and use network resolving and analyzing techniques to accquire the information of key targets attibuting to synergic effects, then at last use methods including plasma resonance and target inhibitions to sttrictly valiate the possible targets, and clarify the targets of baicalin and Panax Notoginsenosidum and their regulatory network. This study may result a breakthrough in searchiing methods for understanding the great connotation of the pharmacological action of Chinese Medicine, and may be significant for clarifying the synergically therapeutic mechanism of Chinese Medicine on AD.

中药组分配伍发挥"多因微效"整体调节作用已有普遍共识，但对其复杂作用机理的本质认识仍处于初级阶段。运用网络药理学的观点和技术，分析与药效相关的体内蛋白质组数据，可以提高查找效率，最大可能地获得靶点蛋白；而应用虚拟识别反向对接靶点自动搜索技术，则能获取与药物结合的蛋白；二者的交集以及蛋白质相互作用网络，可能是多个初级和次级靶点的相互作用、协同制衡产生的集成结果。本研究拟在黄芩苷、三七总皂苷配伍治疗老年性痴呆协同增效等药效学作用明确的基础上，通过网络药理学和虚拟识别反向对接技术，构建药物相关的差异表达蛋白网络和虚拟计算的结构网络，解析协同增效作用的关键靶点信息。最后，通过表面等离子共振及药物靶标阻断等严密的实验手段进行靶点筛选和反向试验验证，最终明确二者作用的靶点蛋白和调控网络。该工作一方面为研究中药机理的丰富内涵提供方法上的新突破，也可能对于阐明中药组分配伍协同治疗AD的机制也具有重要意义。

采用网络药理学方法分析与药效相关的体内蛋白质组数据，可以较大程度地提高查找效率，最大可能地确定靶点蛋白；应用分子反向对接法可获得药物与靶标蛋白的结合情况，结合二者及蛋白质相互作用网络，能够较好地揭示药物与靶标蛋白的相互作用关系。本项目采用采用阿尔茨海默病（Alzheimer’s Disease，AD）细胞、动物模型评价中药黄芩的主要有效成分黄芩苷的AD治疗作用，应用正电子发射断层显像（PET）技术、蛋白质组学、网络药理学、虚拟识别反向对接技术、生物信息学、表面等离子共振技术及药物靶标阻断等方法系统研究了黄芩苷抗AD的作用机制，结果显示黄芩苷对AD细胞模型损伤具有明显保护作用，能够明显改善AD大鼠的认知功能损伤，可明显增强AD大鼠多个脑区的葡糖糖代谢，其抗AD作用与调节多种能量代谢相关蛋白、细胞凋亡相关蛋白、氧化应激相关蛋白密切相关。其中，14-3-3zeta蛋白为差异蛋白调控网络的关键节点蛋白。采用分子对接方法计算出了黄芩苷与14-3-3zeta蛋白的结合域及结合能力。表面等离子共振实验表明黄芩苷与14-3-3zeta蛋白具有较强的结合能力。药物靶标阻断实验结果表明，黄芩苷的神经保护作用与14-3-3zeta蛋白密切相关。该研究结果阐明了黄芩苷抗AD作用的分子机制，明确了其靶点蛋白，对中药黄芩在神经退行性疾病中的应用及AD防治具有非常重要的意义和参考价值。