加味人参乌梅汤调控腹泻大鼠菌群－宿主代谢交互水平衡效应的宏基因组学研究

That bacteria-host interactions balance provides a new strategy for integrated regulation of traditional Chinese medicine. The completed National Natural Science Foundation study shows that modified Renshen Wumei decoction on diarrhea in rat intestinal flora effect together with host metabolic phenotype has a regulatory role, but its mechanism is unknown. Diarrhea is the disease of water metabolism imbalance, while the intestinal microflora has important significance to the host water metabolic balance. Whether modified Renshen Wumei decoction regulating the fluid interaction between the process of flora metabolism and host metabolism to exerts its fluid antidiarrheal effect or not? Recent ground-breaking advances show that flora GABAA-R signaling pathway interacts with the host a total of metabolites are diarrhea disease water metabolism regulation of key targets and a valid path. Accordingly hypothesized that "modified Renshen Wumei decoction regulating the covariant effect of the intestinal flora-host water metabolism interaction balance by GABA-signaling pathway to exerts its fluid antidiarrheal effect”. This subject used strategies and methods， such as intestinal metagenomics , NMR metabolomics, signaling pathway microarray, from the point of the balance of water metabolism mechanism，on the whole, cellular, molecular and gene level, to prove the overall regulation effect of modified Renshen Wumei decoction on the microflora - host water metabolism interactive balance covariation effect by GABA-signaling pathway, further more to enrich the scientific connotation of traditional Chinese medicine therapy with "bacteria group-host interactive".

菌群-宿主交互平衡为研究中医药整合调节作用提供了新策略。前一国家自然基金研究表明，加味人参乌梅汤对腹泻大鼠肠道菌群与宿主代谢表型共变效应具有调控作用，但机制未明。腹泻是水液代谢紊乱疾病，而肠道菌群对宿主水代谢平衡具有重要意义。加味人参乌梅汤是否通过调控菌群与宿主代谢流程中的水液代谢交互发挥益气生津止泻效应呢？新近研究表明，菌群与宿主交互共代谢产物GABAA-R信号通路是腹泻病水液代谢平衡调控的关键靶点和有效路径。据此提出“加味人参乌梅汤通过GABA能信号通路调控菌群-宿主代谢交互水平衡发挥其益气生津止泻作用”假说。课题整合运用肠道宏基因组学、代谢组学、信号通路基因芯片等研究策略和方法，从水液代谢平衡机制切入，在整体、细胞、分子和基因水平，探明加味人参乌梅汤通过GABA能信号通路对菌群-宿主代谢交互水平衡效应的整体调控作用机制，从“菌群-宿主交互”的角度进一步丰富中医药整体调整的科学内涵。

菌群-宿主交互平衡为研究中医药整合调节作用提供了新策略。菌群与宿主交互共代谢产物GABA能信号通路是腹泻病水液代谢平衡调控的关键靶点和有效路径。本课题从水液代谢平衡机制切入，整合运用肠道宏基因组学、代谢组学等研究策略和方法，以探明中药复方加味人参乌梅汤通过GABA能信号通路对菌群-宿主代谢交互水平衡效应的整体调控作用机制。结果提示：①加味人参乌梅汤具有良好的益气生津止泻效应，其中人参-乌梅酸甘相配发挥主要作用。②加味人参乌梅汤有效促进肠道有益菌群（类杆菌纲、乳杆菌属、双歧杆菌属等）的富集，富集的功能类群参与并有效调控GABA代谢途径。③筛选代表性差异代谢物并进行验证，提示加味人参乌梅汤对腹泻模型大鼠血清中的关键代谢物存在良性调节作用。④肠道菌群与血清差异代谢物关联分析显示二者存在显著的相关性，而GABA是菌群与宿主交互共代谢重要介质。⑤加味人参乌梅汤、人参-乌梅药对可通过下调PI3K/AKT/GABAA-R信号通路上游/下游信号IL-1β、GABA、GABAA、PI3K、AKT2等基因和蛋白表达水平，从而发挥其对水液代谢相关因子（Occludin、Claudin-1、ZO-1、MUC2、MUC6、AQP3、AQP8等）的调控而达到益气生津止泻效应。⑥加味人参乌梅汤、人参-乌梅药对可通过下调TLR4/NF-κB信号通路中TLR4、myD88、p-NF-κBp65基因/蛋白表达而减轻炎症免疫损伤，GABA可能通过PI3K/AKt信号通路的下游信号NF-κB信号通路参与了免疫调控作用。本项目研究初步表明，加味人参乌梅汤通过GABA能信号通路调控菌群-宿主代谢交互水平衡而发挥其益气生津止泻效应，其中人参-乌梅酸甘相配发挥核心主要作用。项目立足于“菌群-宿主交互”角度，进一步揭示了加味人参乌梅汤益气生津止泻效应机制，丰富了中医药整体调整的科学内涵。