Metabolism Of Codonopsis Pilosula Saponins By Coexisting Polysaccharides Alleviates Gut Microbial Dysbiosis With Acute Colitis Mice

Inflammatory bowel disease(IBD)mainly includes Ulcerative Colitis(UC)and Crohn's Disease(CD).It is an autoimmune disease caused by genes,metabolism,immunity and other factors.Recovering disorder of gut microbiota is a new research target for the treatment of IBD.Herbal Medicine(HM)as the representative of the supplement and alternative therapy is the key method in the clinical treatment of IBD.Codonopsis pilosula(Franch)Nannf(CPN)serves as a classic HM for replenishing qi.Its representative components,Codonopsis pilosula polysaccharides(Cp)and Codonopsis pilosula saponins(Cs),have a wide range of pharmacological activities,such as anti-inflammatory and immuno-modulate effect.In addition,Cp can be broken down into small molecules under the action of gut microbiota and absorbed by the host to exert its pharmacodynamic effect.But so far,Cp has not been fully utilized in the treatment of IBD.In long-term clinical application,CPN is mainly prepared by decoction and administered orally.Because the complexity of the composition of HM and the synergistic effects of their major chemical components are not yet clear.So it is difficult to find the components that play an effective role in the treatment of disease,and to track the interaction mechanism between the active ingredients.Therefore,it is of great significance to conduct in-depth research on the interactions between the active ingredients of CPN and its interaction mechanism in IBD.In this project,the mouse model of acute colitis was established with 3% dextran sodium sulfate(DSS)induction for 7 days(from the 22 nd day of administration to the 28 th day)after Fro,Cp and Cs were gastric administration with healthy mouse models in 21 days.By evaluating the changes of body weight,colon length,disease pathology index,colon tissue sections,and myeloperoxidase(MPO)in colitis mice,the effects of the three drugs on the macroscopic pathological symptoms of colitis were compared.Using Illumina MiSeq 250 sequence,based on the 16 S V3-V4 variable region of bacteria,the regulation of gut microbiota structure and diversity by different drugs in colitis mice was explored,and the dominant genus in each group after intervention with each drug were analyzed.Gas chromatography(GC)was used to determinethe content of short-chain fatty acids(SCFAs)in feces,and the changes associated with SCFAs-associated genus were searched.ELISA was used to detect the expression of pro-inflammatory and anti-inflammatory cytokines associated with Th17/Treg balance.At the end of the experiment,each experimental group was given 300 mg/kg of Cs once on the 29 th day to collect blood and cecal samples;using UPLC-QTOF-MS technology combined with PKsolver software to analyze the absorption of the highest exposure for monomeric compounds in Cs after the administration of three drugs for prevention.Elucidation that Cp could be used as prebiotics to regulate gut microbiota,which affect the mechanism of Cs metabolism.The results showed Fro,Cp and Cs improved the macroscopic pathological status of mice with DSS-induced colitis.Down-regulated the secretion of pro-inflammatory cytokines:interleukin [IL]-17 A,IL-17 F,IL-6,IL-22,TNF-?,up-regulated the expression of IL-10 and TGF-? in Th17/Treg balance,which has the anti-inflammatory effect.16 SrRNA sequence results showed that the three drugs regulated the structure and diversity of gut mocrobiota in mice with colitis to varying degrees.At the same time,Cp selectively stimulated the growth of three important probiotics,Bifidobacterium spp.,Akkermansia spp and Lactobacillus spp.,reduced the abundance of Desulfovibio spp.,Alistipes spp.,Helicobacter spp.,and restored the gut microbiota in colitis mice.Among them,Fro had the best effect,but there was no significant difference from the Cp group.GC results showed that Fro,Cp and Cs increased the content of SCFAs.Through selective enrichment of Blautia,Prevotellaceae UCG-001,Oscillibacter and Quinella were reported to promote the production of SCFAs,improved the systemic and local energy supply of SCFAs.In addition,the study found after prevention of three drugs for 21 days,by adjusting the imbalance of gut microbiota in mice with colitis,the absorption of the most exposed monomeric compound Cs-415.2 of Cs in colitis mice was enhanced.At the same time,among which,Fro had the best effect,followed by Cp.In summary: Cp had the good prebiotic properties and shown good application prospects in the prevention and treatment of acute colitis.On the one hand,this study confirmed the potential prebiotic effect of Cp.On the other hand,it also revealed the Cp in CPN could affect the metabolic mechanism of Cs in the serum by restoring the disorder of gut microbiota in mice with colitis.To lay the foundation for the development of new micro-ecological preparation by using Cp as a prebiotic.In addition,it also provided a theoretical basis for the discovery of effective substances in HM.