基于肠道微生态学结合代谢组学的黄连解毒汤改善2型糖尿病胰岛素抵抗的药效物质基础及其作用机制研究

项目名称： 基于肠道微生态学结合代谢组学的黄连解毒汤改善2型糖尿病胰岛素抵抗的药效物质基础及其作用机制研究

项目编号： No.81473319

项目类型： 面上项目

立项/批准年度： 2015

项目学科： 医药、卫生

项目作者： 谢智勇

作者单位： 中山大学

项目金额： 68万元

中文摘要： 胰岛素抵抗（IR）是2型糖尿病（T2DM）的核心机制之一。黄连解毒汤能明显改善T2DM-IR。虽然T2DM-IR与肠道菌群关系密切，但鲜有从肠道菌群角度来探讨黄连解毒汤改善T2DM-IR机制的报道。本课题组前期研究发现肠道菌群结构变化是黄连解毒汤改善T2DM-IR的重点环节。因此本项目拟进一步从肠道菌群切入，基于肠道微生态学和代谢组学，利用PCR-DGGE等分子生物学技术和现代分析仪器的代谢组学方法分析T2DM-IR模型大鼠造模前后及黄连解毒汤干预前后的血、尿、粪等的肠道菌群谱和代谢组学谱，结合多变量分析，发现鉴定并定量与改善T2DM-IR关联的代谢物与功能菌，构建黄连解毒汤-肠道菌群-T2DM-IR研究体系，研究其改善T2DM-IR与肠道微生态失衡、平衡间关系，阐明其改善T2DM-IR的药效物质基础及机制，并为研究中医药治疗与肠道菌群相关的肥胖、T2DM等代谢性疾病的机制提供新思路。

中文关键词： 黄连解毒汤；代谢组学；肠道微生态学；药效物质基础；胰岛素抵抗

英文摘要： Huang-lian-jie-du decoction is a very famous and classic decoction which has been used to treat damp-heat syndrome patient for more than one thousand years. Until now, a number of studies have demonstrated that it can obviously improve the insulin resistance in type 2 diabetes mellitus. As insulin resistance in type 2 diabetes mellitus was one of the core pathogenesis, which was closely relevant to gut microbiota. However, the mechanism of Huang-lian-jie-du decoction improving insulin resistance in type 2 diabetes mellitus based on gut microbiota was poorly understood. Preliminary study in our lab has shown that the change of gut microbiota composition was a key issue for the Huang-lian-jie-du decoction improving insulin resistance in type 2 diabetes mellitus. Therefore, the mechanism and therapeutic material basis research of Huang-lian-jie-du decoction improving insulin resistance in type 2 diabetes mellitus will be studied in this project, which will be performed by the combination of 'metabonomics' with 'intestinal microecology' based on the previous research results through the breakthrough point of 'imbalance' or 'balance' of gut microflora. Molecular biology technologies such as polymerase chain reaction-denatured gradient gel electrophoresis (PCR-DGGE) that bases on the 16S rRNA gene, real time polymerase chain reaction (RT-PCR), and metabonomics methods which based on modern analysis instrument will be performed to analysis urine, feces, blood and tissue samples collected from rats. Samples were collected before and after insulin resistance in type 2 diabetes mellitus modeling, before and after Huang-lian-jie-du decoction intervention in nomal and model rats. Through metabonomics and gut microbiota fingerprint data was analysed using SPSS, SIMCA-P+, Matlab, and Cytoscape, the metabolites and functional bacteria which are covariation and correlation each other will be found. Then, the correlative metabolites and functional bacteria will be identified by the high resolution mass spectrometry, cloning and sequencing technology, respectively. The biological basis which relate to relationship of the intestinal microecology 'imbalance' or 'balance' with Huang-lian-jie-du decoction improving insulin resistance in type 2 diabetes mellitus will be well understood. In addition, an evaluation system, 'Huang-lian-jie-du decoction / intestinal microecology and metabonomics / improving insulin resistance in type 2 diabetes mellitus' will be set up. With the use of the evaluation system, the mechanism and therapeutic material basis research of Huang-lian-jie-du decoction improving insulin resistance in type 2 diabetes mellitus will be carried out. At the end, metabonomics in combination with intestinal microecology will provide a new platform for studying the the mechanism about traditional Chinese medicine used to remedy obesity, type 2 diabetes mellitus, and other metabolic diseases which are very closely related to gut microbiota.

英文关键词： Huang-lian-jie-du decoction;Metabonomics;Intstinal microecology;Therapeutic material basis;Insulin resistance