基于内源性代谢产物逆向追踪TRPV1通路后表征辛热（温）药性的生物分 子网络和关键节点

This project is an extension of the National Natural Science Foundation of China. The research group in the implementation of the project found that the thermal effect of TRPV1 channel mediated is the common molecular basis of a variety of herbs. However, the thermal effect of TRPV1 channel and its output is not a one to one direct docking mode, and there are complex downstream molecular pathways and networks post-TRPV1 channel. The conventional single point mode of molecular pharmacology is difficult to find the complex pathway and target that participates in the characterization of heat and hot natures of herbs. Based on this, this project intends to introduce the whole idea of integrated metabolic system with science and technology, with the help of chemometrics methods, focusing on the post-TRPV1 channel neuronal and thermogenesis tissues, we explore the herbal hot/warm pungent nature-related common endogenous metabolites and metabolic spectrum, and then through the analysis of KEGG network module to reversely trace the post-TRPV1 channel key protein nodes and core pathways, and to construct the molecular association pattern of the overall effect of hot/warm herbs, with the aim of comprehensively and systematically revealing the scientific nature of herbs, and thus enriching and perfecting the theoretical connotation of traditional Chinese medicine, and accumulate the rational data to reveal the modern biological basis of the theoretical system of traditional Chinese medicine.

本项目是前期国家自然科学基金的延伸性研究。课题组在实施项目中发现：TRPV1通道介导的热效应是多种辛温（热）中药药性表征的共性分子基础。但TRPV1通道与其输出的终端热效应并非一对一的直接对接模式，其间存在着复杂的下游生物分子通路和网络。传统的单点式的分子药理学模式难以发现TRPV1通道后的参与表征辛热（温）中药药性的复杂通路和靶点。基于此，本项目拟引入具有系统整体理念的整合代谢组学技术，聚焦TRPV1通道后的信号传导和热学变化相关组织，借助化学计量学手段，挖掘辛温（热）中药干预后的共有的内源性代谢产物和代谢谱规律，基于KEGG等网络模块分析逆向追踪TRPV1通道后的与辛温（热）中药药性表征相关的关键分子节点和核心通路，构建辛热（温）中药的整体作用和分子关联模式图，诠释辛热（温）中药药性表征的科学本质，进而丰富和完善中药药性理论的科学内涵，为全面揭示中医药理论体系的生物学基础积累有效数据。

中药药性理论是中医药理论体系的重要组成部分。辛热（温）药性是中药药性的核心内容之一。为探索其生物效应的分子机制，本项目以附子、干姜、肉桂等临床常有的辛热性中药为研究对象，以具有系统整体理念的代谢组学技术手段，关注机体的内源性代谢产物，研究了上述辛热性药物的共有药性特征。通过主成分分析和聚类分析以及网络模块构建等进行了生物学信息的挖掘和抽提，发现了与辛热药性共有生物效应相关的代谢物、靶点和关联通路，并在基因的蛋白表达层面对关键蛋白节点进行了实验验证和分析。总体看，共同差异代谢物有8个，分别为L-缬氨酸、甜菜碱、硬脂酸、花生四烯酸、亚油酸、棕榈酸、反式油酸和油酸，可作为上述热性受试中药相关的生物标志物。利用8个共有差异代谢物构建了代谢网络，共得到10个模块，其中最大的2个模块是花生四烯酸代谢通路和亚油酸代谢通路。高于平均节点度的基因有26个，可作为整个代谢网络中的关键靶点。对这26个基因进行分析，可知它们大多属于细胞色素450超家族。