| Objective: The changes of intestinal flora before and after treatment are detected using 16 s rDNA sequencing technology.The potential target and metabolic pathway of Scutellaria baicalensis in the treatment of inflammatory bowel disease are predicted using network pharmacology.The predicted results of network pharmacology are verified by modern biological techniques,and the molecular mechanism of Scutellaria baicalensis in the treatment of inflammatory bowel disease is discussed.Methods: A mouse model of inflammatory bowel disease was established using chemical reagent induction.After successful modeling,mice were randomly divided into six groups: blank,model,Mesalazine,HQ-L,HQ-M,HQ-H.Those mice were acclimated for 28 d,and intestinal contents of mice were collected before and after treatment.16 s V3V4 region was amplified using 16 s rDNA sequencing technology.The differences of intestinal flora in mice with inflammatory bowel disease before and after treatment with Scutellaria baicalensis were investigate by OTU clustering statistics,alpha diversity,beta diversity and community composition.The effective ingredients and corresponding potential targets of Scutellaria baicalensis were obtained by screening of TCMSP,TCM Database @ Taiwan and other databases.The potential targets for inflammatory bowel disease were obtained by searching OMIM,Drugbank and other databases.The potential targets and related signaling pathways of Scutellaria baicalensis for treating inflammatory bowel disease were revealed by integrating and topological analysis of Cystoscape3.6.0software.Finally,western blot was used to detect the expression of related protein factors in mouse colon tissue before and after treatment with Scutellaria baicalensis.Results: The intestinal flora test results showed that the abundances of Parasutterella,Parabacteroides,Citrobacter,etc.in the model group are reduced significantly,and the abundance of Akkermansia,Saccharibacteria,Acetatifactor,Turicibacter,etc.are increased significantly;The abundances of Saccharibacteria and Acetatifactor in the Scutellaria baicalensis group are decreased significantly,the abundances of Parasutterella andCitrobacter are on the rise,and the abundance of bacteria is proportional to the concentration of Scutellaria baicalensis.21 active components of Scutellaria baicalensis,157 targets for components and 320 targets for diseases are obtained by screening of Pharmacology related databases.29 potential targets for Scutellaria Baicalensis for inflammatory bowel disease are revealed by integrating component targets with disease targets and topological analysis,including RELA,TNF,JUN,IL6,MAPK14,CASP3,IKBKB,etc.148 signal pathways are obtained using KEGG pathway enrichment analysis,including NF-κB,TNF signaling pathways,etc.The expression levels of TNF,JUN,RELA and IL-6 protein in colon tissues were detected by Western blot.Comparing with the blank group,the protein expression in the model group is significantly up-regulated,which is with a significant statistical difference(P<0.01);Comparing with the model group,the expression levels of the Mesalazine and HQ-H are decreased significantly,which is with statistically significant differences(P<0.01);There was no statistical difference in the expression levels of JUN and RELA between the Mesalazine and the HQ-H(P>0.05),and there was a statistical difference in the expression levels of TNF and IL-6(P<0.05);There was a statistically significant difference in the expression levels of TNF,JUN and IL6 between the Mesalazine and the HQ-M(P<0.05),and there was a significant statistical difference in the expression level of RELA(P<0.01);There was a statistically significant difference in protein expression between the Mesalazine and the HQ-L(P<0.01).Conclusion: The effect of Scutellaria baicalensis on intestinal microflora in mice with inflammatory bowel disease and its molecular mechanism was systematically investigated using 16 s rDNA sequencing,network pharmacology and Western blot technology.The results confirm that Scutellaria baicalensis may increase the abundance of beneficial bacteria and reduce the harmful bacteria by adjusting the structure of intestinal microorganisms,thereby relieving inflammation.At the same time,it inhibits the expression levels of RELA,TNF,JUN,and IL6 protein factors in the inflammatory signaling pathway,and finally plays a role in treating inflammatory bowel disease. |
