Study On The Mechanism Of Gegen Qinlian Decoction Based On Gut Microbiota In Model Rats With Acute Enteritis

1 Object In this study,the model rats with acute enteritis were induced by dextran sodium sulfate in order to study the effects of Gegen Qinlian decoction and it's different compatibility on the gut microbiota diversity on normal and model rats and the difference of metabolism of pharmacodynamic components.Combined with the inflammatory index,the compatibility laws of Gegen Qinlian decoction was preliminarily explored from the interaction between gut microbiota and TCM.It was hoped to lay a foundation for further revealing the deep mechanism of action Gegen Qinlian decoction.2 Methods 2.1 Therapeutic Effect of Gegen Qinlian Decoction on Model Rats with Acute Enteritis The concentration of TNF-?,IL-6 and IL-10 was determined by enzyme-linked immunosorbent assay betwteen normal rats,model rats with acute enteritis induced by dextran sodium sulfate and other administration groups.Meanwhile,the therapeutic effect of Gegen Qinlian decoction on acute enteritis was evaluated by combining the scores of colonic injury and partial colonic dryness ratio as well as pathological section.2.2 Effect of Gegen Qinlian Decoction on Intestinal Microbial Diversity in Rats with Acute Enteritis The High-throughput sequencing of 16 S r RNA technology was used to study the effects of the whole decoction group and it's different compatibility groups on the overall gut microbiota composition and proportion of rats with acute enteritis.The different species in the level of phylum and genus were find between Gegen Qinlian decoction and it's different compatibility groups.2.3 Metabolic Differences of Main Components of Gegen Qinlian Decoction by Gut Microbiota The UHPLC-LTQ-Orbitrap-MS was used to analyze the metabolism of flavonoids,saponins and alkaloids in Gegen Qinlian decoction and its different compatibility groups after anaerobic incubation of gut microbiota in vitro.The effects of compatibility on intestinal bacterial metabolism of the main components of Gegen Qinlian decoction were studied combiened with the multivariate statistical analysis.3 Results 3.1 Therapeutic Effect of Gegen Qinlian Decoction on Model Rats with Acute Enteritis The model rats showed weight loss,fecal occult blood,colon edema,colon recess and epithelial structure destruction,which suggested that the model had the pathological characteristics of acute enteritis.The levels of TNF-?and IL-6 in the model group were significantly higher than those in the normal group,while the levels of IL-10 were significantly lower than that in the normal group.Compared with the model group,after the intervention of Gegen Qinlian decoction and its different compatibility the rats hair returned to normal,their weight increased and the injury of colon decreased.What's more,the pathological results showed that the intestinal glands of the therapy groups rats became longer and abundant,and the damage degree of crypt structure in the mucosa was alleviated,bleeding and edema were alleviated,As well as the contents of TNF-? and IL-6 decreased,while the contents of IL-10 increased compared with model rats.3.2 Effect of Gegen Qinlian Decoction on Intestinal Microbial Diversity on Rats with Acute Enteritis The Alpha diversity and Beta diversity showed that there were significant differences in gut microbiota composition of rats in each group.Except the Gegen Gancao group,other groups reduced the increase of abundance and diversity caused by acute inflammation.Bifidobacterium and other beneficial bacteria only appeared in the normal group.Compared with the normal group,Lactobacillus was significantly reduced,Bacteroides,Escherichia-shigella,Clostridium_sensu_stricto₁ were up-regulated in the model group.Compared with the model group,the different compatibility of Gegen Qinlian decoction reduced the abundance of Clostridium_sensu_stricto₁ which related to intestinal inflammatory diseases.The whole decoction group,Gancao-free group and Qinlian groups increased the abundance of beneficial bacteria significantly,such as Akkermansia.The whole decoction group went to the licorice caused the increase of Ruminococcaceae_UCG₀13.The harmful bacteria Desulfovibrio,beneficial bacteria Allobaculum and Bacteroidales_S24-7_ukn were enriched in the Gancao-free group.The Gegen-free group caused the up-regulation of Proteobacteria and Burkhonderiales and decreased the relative abundance of Allobaculum and Bacteroidales_S24-7_ukn.3.3 Metabolic differences of main components of Gegen Qinlian Decoction by Gut Microbiota After the gut microbiota metabolism,five flavonoid components,one saponin component,four alkaloid components and 14 metabolites were identified.The results of multivariate statistical analysis showed that Gegen Qinlian decoction and its composition of prescription were gathered into 5 categories.The ten different components of the classification were identified,and the rules of compatibility on the metabolism of components were preliminarily discussed.4 Conclusion Gegen Qinlian decoction plays therapeutic effects in dextran sodium sulfate-induced acute enteritis model rats by reducing the levels of IL-6 and TNF-?,increasing the levels of IL-10,inhibiting inflammation,alleviating mucosal injury and regulating the diversity of gut microbiota.According to the study of metabolism intestinal bacteria,it is concluded that there is a complex network relationship among diseases,bacteria and drugs,and different composition of prescription can cause abnormal changes of intestinal bacteria and differences in the level of composition metabolism:The anti-inflammatory and mucosal repairing effects were more obvious in the whole decoction group and the the Gancao-free group,Huanglian and Huangqin are the key factors in regulating intestinal bacterial balance in Gegen Qinlian decoction.Their effect on regulating gut microbiota will enhanced with Gegen and Gancao.The metabolism of baicalin and wogonoside in the whole decoction group were more than that in the different compatibility groups,which were beneficial to the exertion of their pharmacodynamics.Most of the alkaloids in Huanglian increased in the intestinal metabolism.Comparing the results of different compatibility groups,it was speculated that Gancao may play an important role in the metabolism of alkaloids.