四磨汤对不同胃肠功能障碍小鼠肠道微生物的影响及其机理研究

The intestinal microbes is a very important part in intestinal microecology, which has a close relationship to many kinds of physiological function, such as human nutrition and immune. The curative effect of Simo Decoction(SD) on different gasastrointestinal dysfunction is distinct. The effect of SD on intestinal microbes and the microecology mechanism of its curative effect has not been reported. The project will construct model mice with chronic stress gastrointestinal dysfunction, constipation with spleen deficiency and constipation with accumulation of dry feces. Then the microorganism culturing technology will be used to analyse the effect of SD on intestinal culturable microbiota. And it will discusses the adjusting action of SD on intestinal microbes. The metagenome DNA of intestinal microbiota will be extracted. The effects of SD on molecular diversity of intestinal microbes will be analysed by modern molecular biology technology, such as T-RFLP. It will explore and analyse the effect of SD on molecular diversity of intestinal microbes. The intestinal microbes which have a relationship to the curative effect will be found out. The effect of SD on gut lactobacillus, which is important gut probiotics, will also be detected by T-RFLP. Finally,the microecology mechanism of SD on different gasastrointestinal dysfunction will be revealsed with its curative effect. The results will clarify the microecology mechanism of SD which is used to treat different gasastrointestinal dysfunction. The research findings and technology system have the great significance in clarifying the microecology mechanism of traditional Chinese medicine's curative effect.

肠道微生物与人体多种生理功能密切相关，便秘等胃肠功能障碍与肠道微生态失调有关。四磨汤对胃肠功能障碍疗效显著，但对肠道微生物的影响及其疗效的肠道微生态学机理未见报道。本项目拟通过动物实验，分别制备慢性应激胃肠功能障碍小鼠、脾虚便秘小鼠及燥结便秘小鼠模型，灌胃给药四磨汤，运用微生物培养技术分析四磨汤对肠道可培养微生物的影响，探讨四磨汤对肠道微生物的调节作用；提取肠道微生物宏基因组，运用T-RFLP等现代分子生物学技术探析四磨汤对肠道微生物分子多样性的影响及其疗效相关肠道微生物；应用T-RFLP技术进一步探究四磨汤对肠道乳酸菌的作用，明确四磨汤对肠道重要有益菌的影响。结合四磨汤对不同胃肠功能障碍小鼠的疗效，揭示四磨汤对不同胃肠功能障碍疗效的肠道微生态机理。研究结果为阐明四磨汤对不同胃肠功能障碍的疗效机理提供实验依据。本项目形成的研究成果与技术体系，对运用现代生物学技术揭示中医药疗效机理意义重大。

肠道微生物与人体多种生理功能密切相关，便秘等胃肠功能障碍与肠道微生态失调有关。四磨汤对胃肠功能障碍疗效显著，但对肠道微生物的影响及其疗效的肠道微生态学机理未见报道。本项目拟通过动物实验，制备功能性胃肠病模型动物，灌胃给药四磨汤，采用传统微生物培养技术、分子生物学技术和生物信息学方法相结合，测定肠道可培养细菌的数量变化、分析四磨汤对实验动物肠道微生物分子多样性的影响，为阐明四磨汤疗效的微生态学机理提供依据。. 本课题结合前期研究工作，制备了胃肠功能障碍小鼠模型，通过灌胃给药，观察四磨汤对胃肠功能障碍动物的疗效，应用微生物培养法研究四磨汤对肠道可培养细菌、真菌（霉菌与酵母菌）、乳酸菌、双歧杆菌、大肠杆菌、肠球菌等肠道菌群的影响，明确了四磨汤在调控肠道菌群中的作用；通过观察四磨汤对胃肠功能障碍动物的疗效，提取肠道宏基因组，应用T-RFLP研究四磨汤对肠道微生物分子多样性的影响，进一步探讨四磨汤对胃肠功能障碍疗效相关的肠道微生物，明确了四磨汤调控胃肠功能障碍肠道微生态的作用及其机理，为四磨汤疗效提供肠道微生态学实验依据；通过观察四磨汤对胃肠功能障碍动物的疗效，提取肠道宏基因组，设计乳酸菌特异性引物，应用T-RFLP研究四磨汤对肠道乳酸菌分子多样性的影响，进一步探讨四磨汤对胃肠功能障碍动物肠道有益微生物的影响，揭示了四磨汤调控胃肠功能障碍肠道微生态的机理。

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