【摘要】 旨在探讨益生菌发酵中草药前后的活性成分变化，挖掘其益生作用机制。利用清热解毒类中药材作为原料，结合产酶乳酸菌(lactic acid bacteria,LAB)和枯草芽胞杆菌(Bacillus subtilis)进行发酵，应用薄层色谱、高效液相色谱、非靶向代谢组学等技术分析中草药发酵前后活性成分以及代谢物的变化。结果:发酵后总黄酮、黄芪总皂苷、总多糖含量分别提升71.3%、33.5%、4.9%，抑菌活性增强;对发酵前和发酵后的中草药进行非靶向代谢组学测定，共检测到2 500个代谢物，多变量统计分析筛选出719个差异代谢物，其中显著差异代谢物有24个显著上调，36个显著下调;对差异代谢物进行京都基因与基因组百科全书(KEGG)富集分析揭示了主要参与的代谢通路，包括次级代谢物生物合成、类黄酮生物合成、嘌呤代谢、黄酮及黄酮醇生物合成、甘油磷脂代谢、花生四烯酸代谢等;相较于未发酵中草药，对发酵中草药中显著上调以及下调的代谢物进行分析发现，上调的代谢物具有抗炎、抗病毒、抗氧化等活性，而下调的代谢物与细胞毒性、神经元功能紊乱相关。综上，中草药发酵可提升有效成分含量，增强抑菌、抗炎、抗病毒、抗氧化活性，同时降低细胞毒性。更多还原

【Abstract】 To explore the changes in active ingredients before and after the fermentation of traditional Chinese medicine by probiotics,and to explore the mechanism of their beneficial effects,a variety of Chinese herbal medicines were used as raw materials,combined with enzymeproducing lactic acid bacteria and B. subtilis for fermentation. Then,thin-layer chromatography,high-performance liquid chromatography,and non-targeted metabolomics were applied to analyze the changes in the active ingredients as well as the metabolites of the Chinese herbal medicines before and after fermentation. The results showed that the contents of total flavonoids,total astragalus saponins and total polysaccharides were enhanced by 71. 3%,33. 5% and 4. 9%,respectively,and the antibacterial activity was enhanced after fermentation. Non-targeted metabolomics was performed on the herbs before and after fermentation,and a total of 2 500 metabolites were detected,and multivariate statistical analysis screened 719 differential metabolites,with 314 significantly up-regulated and 405 significantly down-regulated. KEGG enrichment analysis of the differential metabolites revealed the major metabolic pathways involved,including secondary metabolite biosynthesis,flavonoid biosynthesis,purine metabolism,flavonoid and flavonol biosynthesis,glycerophospholipid metabolism,and arachidonic acid metabolism. Analysis of significantly up-regulated as well as down-regulated metabolites in the fermented herbs,compared with the unfermented herbs,showed that the up-regulated metabolites possessed anti-inflammatory,antiviral,and antioxidant activities,whereas the down-regulated metabolites were associated with cytotoxicity and neuronal dysfunction. In summary,herbal fermentation increased the content of active ingredients and enhanced antibacterial,anti-inflammatory,antiviral and antioxidant activities,and reduced cytotoxicity.更多还原