基于代谢组学的肝郁化火证多维整体性和动态演化性研究

The core feature of syndrome lie in its multidimensional integrity and dynamic evolution, that is, the biological basis of syndrome contains multiple levels/dimensions, and changes consistently over time. Metabolomics has the characteristics of integrity, dynamic and in vivo, which means that it’s supposed to be a unique tool for studying the nature of syndromes. This study is aiming to explain the nature (multidimensional integrity and dynamic evolution) of a syndrome called liver depression transforming to fire(LDTF). We intend to study the same syndrome (LDTF) which is found commonly in 2 different diseases (hypertension and anxiety). The analysis technology of metabolomics UPLC-MS / MS and a multi-time point schedule will be adopted. By focusing on the mechanism of psychological stress and inflammation, related upspring gene and protein need further confirmed. This project will provide a scientific evidence for the biological basis LDTF, but also a new method for the modernization of TCM syndrome.

证候复杂特征的核心在于多维整体性和动态演化性，即证候的生物学基础包含多个层面、多个维度，且随时间的推移不断发生演化。代谢组学具有整体、动态、在体的特点，对于揭示证候本质具有独特的优势。本课题以阐释肝郁化火证候本质为研究目标，以该证的多维整体性与动态演化性的生物学基础为研究内容，以异病（高血压和焦虑症）同证（肝郁化火证）病证结合模型为研究对象，采用多时点采集研究方案，运用代谢组学UPLC-MS/MS等分析技术寻找标志性代谢物，并从心理应激与炎症角度入手，并追溯标志性代谢物上游对应的蛋白质和基因进一步验证。该课题预期目标的实现将为肝郁化火证的现代科学内涵提供科学依据，同时为中医证候现代化研究提供一种新的尝试。

肝郁化火证广泛存在于临床各科，其生物学基础仍需探讨。本研究将肝郁化火证的多维整体性和动态演化性为研究重点，借助非靶向代谢组学技术手段，在分别建立高血压/焦虑肝郁化火病证结合大鼠模型的基础上，运用超高效液相色谱-串联四极杆/飞行时间质谱和多元统计分析方法：对模型动物血浆进行代谢物组检测和分析，提取同证异病的共性差异代谢物，发现乙酰肉碱、犬尿氨酸、N-乙酰谷氨酸、吲哚、5-羟基吲哚乙酸、神经酸6个与该证密切相关的潜在标志性代谢物；通过鉴定分析脑组织差异代谢物及差异表达基因，初步建立简单的基因-代谢网络；分时点检测成模过程中代谢物改变，发现证候形成过程中代谢物总体特征随时间演变，其中5-羟基吲哚乙酸的含量改变明显，可作为该证形成过程中的潜在标志性代谢物。本研究为肝郁化火证的现代科学内涵提供科学依据，同时其他证候的代谢组学研究提供了借鉴。