肠道微生态与代谢组相关联的大枣补脾益胃功效物质及作用机制研究

Traditional Chinese medicine (TCM) believes that Jujubae Fructuw has the effect of reinforcing spleen and stomach, and commonly used in prescription and health care, which have been confirmed by the reports and our previous studies. Modern research shows that spleen deficiency is closely related to alterations in the intestinal microflora. Accordingly, based on the Chinese medicine theory, the project proposed to study the bioactive components and molecular mechanism of Jujubae Fructus for reinforcing spleen and stomach from intestinal microecology, host metabolome, and their interrelation. The following works will be carried out: (1) Based on the molecular biology techniques, an intestinal microflora fingerprint of spleen deficient animal models will be established, and the interaction between the chemical composition of Jujubae Fructus and animal models' intestinal flora will be studied; (2) The metabolite profiles of spleen deficient animal models will be established using the metabolomics methods and techniques, and the intervention on the biomarkers and metabolic pathway by the chemical composition in Jujubae Fructus will be studied; (3) Based on multivariate statistics methods, the co-variation between intestinal flora fingerprint of animal models and host metabolic fingerprint will be elucidated and the potential relation between intestinal flora and host metabolic phenotype, as well as the intervention of each component in Jujubae Fructus will be investigated. The final objects of these studies are to systematically clarify the bioactive components and biological mechanisms of Jujubae Fructus for reinforcing spleen and stomach effects and to reveal the scientific connotation of its invigoration effects.

传统中医认为大枣具有补脾益胃之功效，并常用于方剂配伍和养生保健。前人报道及本课题组前期研究均证实其补脾益胃功效确切。现代研究显示脾虚证与肠道微生态改变密切相关。据此，本项目在中医药理论指导下，提出基于肠道微生态、宿主代谢组，以及二者相互关联的角度，开展大枣补脾益胃功效物质基础及作用机制研究，拟开展工作如下：（1）基于分子生物学技术，构建脾虚模型动物肠道菌群结构指纹谱，研究大枣化学组分与肠道菌群的相互作用；（2）应用代谢组学的研究方法与技术，构建脾虚模型动物代谢指纹谱，研究大枣化学组分对代谢物谱的干预效应及其调控靶标、代谢通路等；（3）基于多变量统计方法，对脾虚模型动物肠道菌群指纹谱与宿主代谢指纹谱进行共变异分析，挖掘肠道菌群/株与模型动物代谢产物之间的潜在联系，并研究大枣各组分的干预作用。以期较为系统地回答大枣发挥补脾益胃功效的物质基础与生物学机制，揭示大枣药食兼用及其补益功效的科学内涵。

大枣自古就有“补脾益胃”功效的记载和临床应用，早在《黄帝内经·素问》即有“枣为脾家之果”的论述。长期以来，大枣也是诸多补脾益胃名方中常用的配伍用药之一。现代研究证实大枣补脾益胃功效确切，且具有可减缓药性峻烈药物对胃肠道及机体正气的损伤作用，具有顾护脾胃之功效。.本项目在对大枣传统功效作用特点进行梳理分析的基础上，构建了基于肠道微生态与宿主代谢组相关联的大枣补脾益胃功效物质基础及作用机制研究方法。应用代谢组学研究方法阐明了大枣提取物及各化学组分可干预的脾虚模型动物代谢物谱效应变化、调控的潜在生物标志物群、影响的代谢通路，并经Western blotting证实大枣补脾益胃功效的发挥与其各化学组分不同程度地调节PLA2-COXS/LOXS通路和TLR4/NF-κB通路蛋白的表达密切相关。采用16S rDNA测序技术对脾虚动物模型肠道菌群结构进行研究，结果显示：与空白组比较，脾虚模型组大鼠粪便中门、纲、目、科、属分类水平上均发现差异菌群，大枣提取物可不同程度地改善模型动物不同肠段差异菌属相对水平。肠道菌群结构与代谢物谱共变异分析结果显示，脾虚模型动物肠道菌群中多个差异菌属与宿主代谢差异物表现出较高的相关性，提示其在脾虚证的发生发展中呈现出一定协同作用。在此基础上，拓展性开展大枣缓和药性作用研究，证实大枣与巴豆霜配伍应用可在血清免疫指标、肠道菌群结构等方面表现出一定的配伍减毒和缓解峻猛之性作用。药代动力学研究结果显示，大枣三萜酸类成分在模型动物体内代谢动力学参数与正常动物具有显著差异。本项目从肠道菌群结构与宿主代谢组角度较为系统地回答了大枣发挥补脾益胃功效的物质基础与生物学机制。.项目研究成果在国内外发表学术论文16篇，其中SCI源期刊收录6篇，申请国家发明专利4项，其中获授权2项；研究成果获国家科技进步二等奖1项（排名第八），省部级科技进步一等奖1项（排名第三）。项目负责人郭盛入选江苏省“六大人才高峰”高层次人才培养对象、江苏省“双创计划”（科技副总）人才计划。