Study On The Effect Of Berberidis Radix On Mice With Ulcerative Colitis Based On Metabolomics And Intestinal Microflora

Objective: To observe the effect of Berberidis radix on DSS-induced ulcerative colitis in mice and preliminarily explore its mechanism from the perspective of regulating endogenous metabolites and intestinal flora diversity.Methods:(1)60 c57BL/6J mice were randomly divided into control group,model group,mesalazine group as well as other kinds of groups with low,medium and high dose of Berberidis radix,with 10 in each.The control group drank water freely,and the remaining groups drank 2.5% DSS solution continuously for 7 days to replicate the UC model related to mice.From the first day of modeling,each group was given corresponding drug intervention according to 0.2ml/10 g gavage volume,and the control group and model group were treated with equal volume of distilled water gavage.Body weight and disease activity index scores of mice were observed and recorded daily.After collecting the serum,caecum contents and colon of mice on the11 th day,the colon length of each group of mice was recorded.Furthermore,the pathological changes of colon were observed by hematoxylin-eosin staining.The content of TNF-α,IL-6 and IL-10 in mouse tissues were detected by ELISA.(2)The level of endogenous metabolites in serum and feces of mice was detected by ultra-high performance liquid chromatography-time-of-flight mass spectrometry technology.After noise reduction,alignment,matching and normalization of the obtained mass spectrometry data,the differential metabolites detected were characterized by principal component analysis and orthogonal partial least squares discriminant analysis.According to the reference indicators including VIP>1,FC>1.5 or FC < 0.67,and P < 0.05,potential differential metabolites and possible metabolic pathways were screened out. (3)The total DNA of intestinal flora was amplified by PCR with 16 S r RNA encoding gene V3-V4 region as the target sequence.In terms of the community structure,alpha diversity,and beta diversity,the sequencing data obtained from the Illumina platform were utilized to analyze the effects of Berberidis radix on the composition and diversity of intestinal microflora in UC mice.Results:(1)Compared with the model group,the weight of mice in the mesalazine group and three types of Berberidis radix groups increased significantly(P<0.05).It can be seen from HE staining results that the intestinal mucosa of control mice was intact and goblet cells differentiated normally.In the model group,the colonic tissue structure basically disappeared,with congestion and edema in the mucosa and inflammatory cells diffusely infiltrated,and goblet cells were significantly deformed and decreased.Through comparison with the model group,the structure of the mesalazine group and the middle and high dose groups of Berberidis radix was more complete,with less inflammatory cell infiltration and more complete goblet cell structure.(2)The results of metabonomics PCA manifested that each group was well separated under both positive and negative ion modes.Specifically,the serum metabonomics showed that 33 differential metabolites and 23 differential metabolites were identified under the positive and negative ion modes,respectively.Additionally,the results of stool metabolomics revealed that 8 differential metabolites and 35 differential metabolites were determined in the positive and negative ion modes,respectively.The above 99 differential metabolites were introduced into Metabo Analyst 5.0 for pathway enrichment analysis.It was found that the most influential factors were the biosynthesis of phenylalanine,tyrosine and tryptophan,linoleic acid metabolism,phenylalanine metabolism and glycerol phospholipid metabolism.(3)The 16 S r RNA gene sequencing results suggested that the intestinal flora diversity of mice in the model group decreased,along with the disordered flora structure by contrast with the control group.After the intervention of Berberidis radix,the recovery of the diversity and structure of intestinal flora in mice was promoted.Compared with the model group,the relative abundance of Campylobacter,Firmicutes was significantly increased(P<0.05).At the genus level,the main performances were a significant increase in the abundance of Helicobacter(P < 0.05),and an obvious decrease in the relative abundance of Odoribacter,Alistipes,and Rikenellaceae_RC9_gut_group(P<0.05).Conclusion: Berberidis radix can significantly improve the symptoms of mice with ulcerative colitis induced by DSS,and one of its mechanisms may be the regulation of the diversity and structure of endogenous metabolites and intestinal flora.