基于配位化学对中药紫花地丁“清热解毒”的基础研究

The subject is based on the TCM basic principle "Clear away Heat and Toxins" and the most useful Chinese medicine with best "Clear away Heat and Toxins" function is chosen as research object. Under the guidance of Chinese Medicine Coordination Chemistry and combined with our early research, a hypothesis is put forward as "effective parts in Viola Herba (flavonoids ingredients) coordinated with immune adjustment trace elements like Zn, Cu to adjust the immune function of the body", so as to enhance its function of clearing away the heat and toxins..Started from correlation between the form of trace elements and the natures of Chinese Medicine, LC-ICP-MS spectrometry technology will be used to separate formation complexes formed by combination of flavonoids ingredients and immune adjustment trace elements like Zn, Cu. The separated formation complexes will be used to screen the activity inside and outside of body, and the configuration of formation complexes with activity will be identified. Then alveolar macrophages model of RAW264.7 will be chosen to study the effect of active formation complexes on TLRs /NF-κB immunomodulatory signal transduction pathway, so as to find the functional target of them. So the material base and function mechanism of "Clear away Heat and Toxins" efficacy strengthen of Violae Herba can be explored from the molecular level, and the scientific connotation of the TCM basic principle "Clear away Heat and Toxins" based on the Coordination Chemistry theory can be improved, which can also provide a new idea and new method for the material base and function mechanism research of traditional Chinese medicine.

本课题立足于中医"清热解毒"基本治则，以"清热解毒之要药"紫花地丁为研究对象，在中药配位化学学说指导下，结合前期研究，提出假说：紫花地丁可能是通过其有效部位（黄酮类成分）与Zn、Cu等免疫调节微量元素的配位结合，调节机体免疫功能，从而增强其祛"热邪"清"热毒"的功效。.从微量元素形态与中药性效相关性入手，首先采用LC-ICP-MS联用技术分离紫花地丁中黄酮类成分与免疫功能调节剂-Zn、Cu等元素形成的配合物，进行体内、体外活性筛选，并鉴定活性配合物构型；然后以肺巨噬细胞RAW264.7模型探讨活性配合物对TLRs /NF-κB免疫调节信号转导通路的影响, 探寻配合物作用靶点，从分子水平上阐释立足免疫调节的紫花地丁"清热解毒"功效增强的物质基础及作用机理，完善基于"配位化学学说"的中医"清热解毒"基本治则的科学内涵，为中药药效物质基础及作用机理研究提供一种新思路与新方法。

本课题立足于中医“清热解毒”基本治则，以“清热解毒之要药”紫花地丁为研究对象，在中药配位化学学说指导下，结合前期研究，提出假说：紫花地丁可能是通过其有效部位（黄酮类成分）与Zn、Cu等免疫调节微量元素的配位结合，调节机体免疫功能，从而增强其祛“热邪”清“热毒”的功效。.课题首先采用ICP-MS法对不同产地紫花地丁中微量元素含量和水煎液中形态进行分析，并根据实验结果，采用化学分离或合成法对可能存在的配合物进行分离/合成，运用NMR、IR、UV等手段进行结构鉴定。以体内外试验对合成的配合物进行免疫活性筛选，最后以肺巨噬细胞RAW264.7为模型，从TLRs / NF-κB信号通路探讨活性成分免疫调节作用机制。结果表明：紫花地丁中微量元素的含量由高到低依次为Ca>Mn>Zn>Cu>Pb>As>Cd>Hg，其中Mn、Zn、Cu主要以有机态（配合物）形式存在于水煎液中。采用化学合成法，对紫花地丁中黄酮类化合物的Mn、Zn、Cu配合物进行合成，成功合成并推测出6个黄酮类配合物，经过体内外活性筛选，柚皮素锌、柚皮素铜具有显著免疫增强作用。目前，该两个活性成分免疫调节作用机制实验正在进行中。