基于质谱探针技术的模型鼠粪便代谢物全分析方法学研究

肠道菌群组成的结构和比例与人体的健康状况有着密切联系。粪便代谢组分析作为一种相对全面、连续动态、无刺激的研究策略，能够系统客观地反映肠道菌群的代谢物组分及浓度，展示肠道菌群的代谢状态。然而，目前对粪便代谢物的了解十分有限，制约了对肠道菌群的深入研究。本项目以模型鼠粪便代谢组全分析为目标，将代谢物按官能团分为亚代谢组（羧基类代谢物、氨基类代谢物、羰基类代谢物、羟基类代谢物、巯基类代谢物等），通过设计和合成高效的质谱探针并分别标记含有不同官能团代谢物，建立基于质谱探针技术的代谢组的色谱-质谱分析方法，对模型鼠粪便进行亚代谢组分析；在综合不同官能团代谢物分析结果的基础上，实现模型鼠粪便代谢物全分析。希望通过本项目的研究，建立模型鼠粪便代谢组数据库，为深入研究肠道菌群与宿主之间的交互作用，揭示肠道菌群与机体的健康和疾病之间的关系提供重要基础数据和研究手段。

The composition and proportions of human gut microbiota are closely correlated to human health. Since fecal metabolome can neutrally reflect the composition and concentrations of metabolites of gut microbiota in a comprehensive, dynamic, and non-invasive manner, analysis of fecal metabolome can be used to monitor the conditions of gut microbiota. However, comparing with the large number of metabolites discovered in human species so far, the current identified metabolites in faeces of mammalians are still limited. In addition, no fecal metabolites database has been established, which thus greatly restricts the in-depth study of gut microbiota. In this project we focus on the comprehensive analysis of fecal metabolome of model rat by establishing mass probe-based metabolomic analytical strategy. Briefly, the metabolites of fecal metabolome will be divided into sub-metabolomes, including carboxy-containing metabolites, amino-containing metabolites, carbonyl-containing metabolites, hydroxy-containing metabolites, and thiol-containing metabolites, etc. Then we will design and synthesize highly effective mass probes that are used to label sub-metabolomes carrying different functional groups. The labeled metabolites will be further subjected to enrichment and liquid chromatography-mass spectrometry analysis. By integrating the results from the analysis of different sub-metabolomes with our developed mass probe-based strategy, we can establish the comprehensive spectrum of fecal metabolome and construct the model rat fecal metabolome database. Through the comprehensive analysis of fecal metabolome, this project will provide important insight and analytical tools on the elucidation of the interaction between gut microbiota and host’s health and diseases.