青礞石中金属离子对于中医痰证、癫痫的干预作用及药效物质基础研究

Chloriti Lapis(CL) used in Chinese medicine is accurate to the phlegm syndrome and epilepsy clinical curative effect, its chemical composition is inorganic elements, and mainly exists in the ion form. The pathogenesis of phlegm syndrome and epilepsy are closely related ion concentrations inside and outside cells, ion channel, metalloproteinase activity. On the basis of preliminary studies and according to the chemical composition characteristics of CL and the pathogenesis of epilepsy and phlegm, We proposed the hypothesis that CL active material foundation possibly is in the disturbing ion concentration inside and outside the cell, the ion channel, or through affected the ion second messenger which activated cell factor and the ion channel related phlegm syndrome and epilepsy, including further activated the related enzyme after the metal protein union. CL inside the body of various ions relative content, valence, and protein-bound State is the key aspect of its display effect. This subjects use of modern inorganic elements and their morphological analysis technology, the serum pharmacochemistry of TCM and metabolomics combined method, research CL ions relative concentration of in animals, valence and state with metal-protein binding, and associated with the effect to revealing its material basis for the effect. Studies the influence of biomarker group, the pathways, metabolism circuit which relate to phlegm syndrome and epilepsy because of the correlation ion as the second messenger. Revelation CL function mechanism which acting on phlegm syndrome and epilepsy. The study of the subject will provide proper approach and evidence on further mineral-derived TCM investigation.

青礞石对痰证、癫痫临床疗效确切，其化学成分为无机元素且主要以离子形式存在。癫痫、痰证的发病机制与细胞内外离子浓度、离子通道、金属蛋白酶的活性密切相关。鉴于前期研究，根据青礞石化学组成特点，基于痰证、癫痫的发病机制，提出其作用物质基础可能是干扰体内细胞内外离子浓度、离子通道，或通过离子的第二信使作用，激活了与治疗痰证、癫痫作用相关的细胞因子和离子通道，包括与金属蛋白结合后进一步激活相关酶；其进入体内各种离子的相对含量、价态、与蛋白结合的状态为其发挥效应的关键因素。本课题利用现代无机元素及其形态分析技术、血清药物化学和代谢组学相结合的方法，研究青礞石进入动物体内各离子的相对浓度、价态及与金属蛋白结合情况，并与药效相关联，揭示其效应物质基础；研究相关离子作为第二信使对痰证、癫痫的生物标志物群、代谢途径、代谢通路等的影响，揭示青礞石对痰证、癫痫作用的效应本质。本课题为矿物药的深入研究提供了思路。

青礞石1963版《中国药典》即收载用于临床治疗痰证、癫痫等，且疗效确切。然而多年来，其作用机制及效应物质基础仍不明确。鉴于前期研究，深入开展青礞石对痰证、癫痫作用的效应物质基础研究：. 1. 青礞石治疗中医痰证、癫痫动物模型的制备. 根据青礞石的功效特征及其悠久的临床应用经验，最终确定采取了烟熏结合细菌感染复合型的建模方法，建立AECOPD痰热证大鼠动物模型。癫痫发病具有突发性、间歇性，本研究选择制备PTZ点燃癫痫大鼠模型，更符合青礞石平肝镇惊的功效及癫痫发狂、惊风抽搐的临床表现。. 2. 青礞石治疗中医痰证、癫痫的药效学研究. 通过对不同组别大鼠肺组织中NF-κB、IL-8、TNF-α、ICAM-1、MCP-1等，血清中CRP、IL-8、TNF-α、LTB4、 MMP-9、TIMP-1等的含量研究表明，青礞石药材、水煎液和药渣均能改善AECOPD模型大鼠肺部病理改变、降低血清及肺中炎症因子含量，促进气道修复，起到良好的药效作用。青礞石各组均能降低PTZ点燃大鼠海马区的病变程度、提高T-SOD活力、降低MDA含量、升高Na+，K+-ATPase及Ca2+，Mg2+-ATPase活性、降低nNOS的蛋白表达、降低大鼠脑内兴奋性氨基酸递质含量及有效调节皮层及海马区Gly的含量。. 3. 青礞石治疗中医痰证、癫痫的物质基础研究.通过测定AECOPD大鼠血浆、肺组织中多种金属元素含量，分析其具有显著差异的金属元素，并进行关联分析，结果表明，青礞石中Fe、Ti、V、Cu、Zn、Sn、Mn、Co、Mg、Al等金属元素群的相对含量、存在形式可能是其治疗AECOPD的效应物质基础。测定PTZ点燃癫痫大鼠血浆及脑组织中多种微量元素的含量，结果青礞石中Fe、Ca、K、Cd、Zn、Co、Sb、Pb、Ti、Li等金属元素群为其治疗癫痫的效应物质基础。. 4. 青礞石治疗中医痰证、癫痫的作用机制研究.建立了AECOPD大鼠血浆、尿液及肺组织的代谢指纹图谱，共鉴定出30种差异代谢物，影响最为重要（Pathway impact>0.1）的代谢通路共5个。对PTZ点燃大鼠的血浆、尿液及脑组织进行代谢组学研究，共鉴定出22个差异代谢物，影响重要的（Pathway impact>0.1）代谢通路6个。