苦寒中药对大鼠胃肠道菌群结构的影响及其与粪便上清/血清代谢产物的相关性研究

This subject choose 7 kinds of Bitter Cold Chinese material medica such as radix et rhizoma etc., conventional prepared water extracts , taking drugs to rats by gavage using small, medium and large doses respectively, to prepare the rat intervention model by Bitter Cold Chinese material medica through oral administration. At different time points at the 1st, 2nd and 3rd week after the administration, collect the samples of different intestinal segments of the experimental rats, that is stomach, duodenum, jejunum, ileum and colon (including intestinal contents), and the samples of fresh faeces and serum, using polymerase chain reaction - denaturing gradient gel electrophoresis (PCR-DGGE) strategy to test and analyze the fingerprints of gastrointestinal bacterial community and its clustering feature of principal members by real-time quantitative PCR technique to test the quantity of the main dominant microflora in the gastrointestinal tract, and using ultra-high performance liquid chromatography / mass spectrometry (UPLC / MS) system to test and analyze the characteristics of metabolomics in the stool supernatant and serum, and screening of potential biomarkers. So as to study the effect on the gastrointestinal bacterial community of the experimental rats by oral administration of Bitter Cold Chinese material medica and it's correlational study on the metabolin of stool supernatant and serum.The results can provide a scientific basis to evaluate the pharmacological effects and toxicity of the Bitter Cold C Chinese material medica based on the gastrointestinal bacterial community, to reveal the relation involving the theory of bitter and cold damaging the stomach with gastrointestinal bacterial community, and to guide the proper usage of the Bitter Cold Chinese material medica in clinical, and also can do some foundation works of the evaluation system to evaluate the pharmacological effects and toxicity of Chinese material medica based on the intervention effect on the gastrointestinal bacterial community.

选择大黄等7味常见苦寒中药，常规制备水提物，分别以大、中、小不同剂量灌胃给药，制备苦寒中药口服干预大鼠模型。在给药第1、2、3周不同时间点，收集大鼠胃、十二指肠、空肠、回肠和结肠等肠段（含肠内容物）以及新鲜粪便、血清等样本，采用聚合酶链反应-变性梯度凝胶电泳（PCR-DGGE）技术分析胃肠道菌群指纹图谱及其主成分聚类特征，采用实时荧光定量PCR技术分析胃肠道主要优势菌群数量，采用超高效液相色谱/质谱联用（UPLC/MS）技术分析粪便上清和血清代谢组特征，并筛选潜在生物标记物，从而研究口服苦寒中药对实验大鼠胃肠道细菌群落结构的影响及其与粪便上清和血清中代谢产物的相关性。研究结果可为评价苦寒中药基于胃肠道菌群结构的药理效应和毒性作用、揭示中药"苦寒伤胃"是否涉及胃肠道菌群改变及其规律、指导临床合理使用苦寒中药等提供科学依据，为探索基于胃肠道菌群干预效应的中药药效和安全性评价体系积累研究基础。

细菌群落的生理性组合是胃肠道微生态平衡的基本要素，能发挥极其重要的生理作用；当各种因素引起胃肠道细菌群落出现病理性组合时，导致胃肠道微生态失调，体内相应的代谢过程和代谢产物也随之发生改变，并与一系列疾病的发生或/和加重密切相关。苦寒中药作为最常用的中药种类，其基于胃肠道菌群生态的药理效应值得探索，同时其“苦寒”损伤脾胃的问题也不容忽视。本项目根据胃肠道菌群的生理效应及其与中医脾胃功能的密切联系，根据“苦寒伤胃”的基本理论和客观存在，采用基于16S rRNA的分子微生态学技术和基于UPLC/MS的代谢组学技术，考察7味常见苦寒中药对实验大鼠胃肠道细菌群落结构的影响，并进一步分析这种影响与代谢组的相关性，筛选其潜在生物标记物，旨在揭示苦寒中药-胃肠道菌群结构-代谢产物的关联规律。其研究结果可为考察苦寒中药对胃肠道菌群结构的影响及其规律、为评价苦寒中药基于胃肠道菌群结构的药理效应和毒性作用、为苦寒中药的合理应用等提供科学依据，并为进一步探索中药-胃肠道菌群结构-代谢组的药理毒理评估模式积累研究基础。