黄连-吴茱萸药对改变肠道菌群结构与抗溃疡性结肠炎作用的相关性及其机制研究

Coptis-evodia herb couple，widely used to treat 'Pi-wei' related diseases more than one thousand years， has very strong curative effects on Ulcerative colitis. However, the mechanism of action on Ulcerative colitis, the therapeutic material basis and the 'Han-re' compatibility mechanism of Coptis-evodia herb couple is still a puzzle and focus. A number of studies have demonstrated that there was a close relationship between Ulcerative colitis and gut microbiota. Meanwhile, Preliminary study in our group has shown that the change of gut microbiota composition was one of key issues for the curative effects on Ulcerative colitis of Coptis-evodia herb couple. In order to reveal how Coptis-evodia herb couple affect gut microbiota composition in nomal and Ulcerative colitis model rats, our lab will continue to study the intestinal microecology and intestinal metabonomics characteristics using molecular biology technologies such as Polymerase Chain Reaction-Denatured Gradient Gel Electrophoresis (PCR-DGGE) based on the 16S rRNA gene, Real Time Polymerase Chain Reaction (RT-PCR) and metabonomics methods such as UPLC-Q-TOF-MS, GC-MS, NMR, and so on, respectively. Also, the effects of Coptis-evodia herb couple on Ulcerative colitis will be evaluated. The relationship between the change of gut microbiota composition and the effects of Coptis-evodia herb couple on Ulcerative colitis of will be explicitted. Then significant different metabolites and intestinal bacteria will be found, recognized, identified and quantified. Furthermore, an evaluation system, 'ratio of Coptis to Evodia herb couple/ time / effects' of 'Coptis-Evodia / gut microbiota / Ulcerative colitis', will be set up after elucidating and interpreting biological significance of the significant different metabolites and intestinal bacteria, which will applied to elucidate the mechanism of action on Ulcerative colitis, therapeutic material basis and 'Han-re' compatibility mechanism of Coptis-evodia herb couple. Finally, a new method，which is the combination of 'metabonomics' with 'intestinal microecology', correlated between gut microbiota and intestinal metabolites will be set up to understand the complex compatibility of mechanism of tranditinal Chinses medicine using to prevent and treat 'Pi-wei' related diseases which are very closely related to gut microbiota.

黄连吴茱萸药对寒热配伍广泛应用于与中医范畴"脾胃"相关的疾病，抗溃疡性结肠炎（UC）作用显著，而其作用机理及寒热配伍机制一直是中医药研究的困惑与焦点。UC的发病及治疗与肠道菌群结构变化密切关联。本课题组前期研究发现肠道菌群结构变化是黄连吴茱萸药对抗UC作用的关键环节之一。为此，本项目拟围绕肠道菌群，利用现代分析仪器的代谢组学方法和16S rRNA基因PCR-DGGE等分子生物学技术分析UC模型大鼠造模前后及经黄连吴茱萸药对干预前中后的肠源性代谢组学与肠道微生态学特征，结合抗UC作用评价，研究药对改变肠道菌群结构与抗UC作用的相关性，发现识别鉴定并定量相关联的差异代谢物与差异肠道菌，诠释其生物学意义，构建"黄连吴茱萸-肠道菌群-UC"的"配比-时-效"评价体系，阐明黄连吴茱萸药对抗UC作用机理、药效物质基础及寒热配伍机制，为中医药防治与肠道菌群相关的中医范畴"脾胃"疾病的机理研究提供新方法。

黄连吴茱萸药对寒热配伍广泛应用于与中医范畴"脾胃"相关的疾病，抗溃疡性结肠炎（UC ）作用显著，而其作用机理及寒热配伍机制一直是中医药研究的困惑与焦点。UC的发病及治疗与肠道菌群结构变化密切关联。本课题组前期研究发现肠道菌群结构变化是黄连吴茱萸药对抗UC作用的关键环节之一。为此，本项目拟围绕肠道菌群，利用现代分析仪器的代谢组学方法和16S rRNA基因的肠道微生物组学等分子生物学技术分析UC模型大鼠造模前后及经黄连吴茱萸药对（黄连：吴茱萸=6：1和黄连：吴茱萸=1：6）干预前中后的肠源性代谢组学与肠道微生态学特征，结合抗UC作用评价，研究药对改变肠道菌群结构与抗UC作用的相关性，发现识别鉴定并定量相关联的差异代谢物与差异肠道菌，UC模型大鼠的肠道菌群结构及其代谢功能都发生了明显的改变，主要表现为生物多样性降低及有害菌与有益菌的比例失常。左金丸和反左金丸干预均能抑制某些致病菌或条件致病菌（如黏蛋白降解菌Mucispirillum）的增加，同时增加菌群中益生菌（左金丸增加短链脂肪酸产生菌、反左金丸可增加乳酸杆菌）的丰度。左金丸和反左金丸干预都能显著上调在模型组尿样中减少的代谢物，如1, 3-二羟基丙酮、乙酰胺、顺乌头酸、乙醇酸、N-三甲基赖氨酸，同时减少在模型组尿样中上调的腺嘌呤、二甲胺和二甲基亚砜。UC大鼠及左金丸和反左金丸干预后显著变化的代谢通路包括丙酮酸代谢、三羧酸循环、糖酵解与糖异生等糖代谢或能量代谢，甘油酯代谢、丁酸甲酯代谢、酮体的合成与降解等脂质代谢，以及苯基丙氨酸、酪氨酸、色氨酸、缬氨酸等多种氨基酸合成与代谢，初步诠释其生物学意义，并构建"黄连吴茱萸-肠道菌群-UC"的"配比-时-效"评价体 系，初步黄连吴茱萸药对抗UC作用机理、药效物质基础及寒热配伍机制等进行了阐明，为中医药防治与肠道菌群相关的中医范畴"脾胃"疾病的机理研究提供新方法。