IDDF2021-ABS-0196 The effect and immune cell analysis of clostridium butyricum on dextran sulphate sodium induced colitis in mice pretreated with antibiotic cocktail

Background

Inflammatory bowel disease is a chronic inflammatory disease characterized by recurrent abdominal pain and diarrhea, and gut microbiota is one of its causes. Probiotics, as one of the intervention methods of microbiota, play an important role in disease treatment. This study aims to observe the effect and immune mechanism of Clostridium butyricum MIYAIRI 588 (CBM) and it's supernatant on colitis mice which pretreated with antibiotic cocktail (ABx).

Methods

Twenty four Balb/c mice aged 6-8 weeks were randomly divided into four groups, and CBM and its supernatant was used to treat colitis mice which pretreated with ABx. The body weight, disease activity index (DAI) and colon length of mice were observed respectively. The relative expression of inflammatory cytokines in colon tissue was detected by qPCR. And the number of neutrophils, macrophages, Th1 and Th17 cells in lamina propria of mucosa was detected by flow cytometry to observe the changes of pro-inflammatory immune cells in mouse mucosa.

Results

The DAI of mice were decreased, the shortened colon length was improved after CBM and its supernatant intervention (IDDF2021-ABS-0196 Figure1A. Clostridium butyricum and its supernatant ameliorated colitis and regulated immune cells; DAI scores of mice after C. Butyricum gavaging with antibiotic cocktail pretreatment at the end of experiment)( IDDF2021-ABS-0196 Figure1B. Clostridium butyricum and its supernatant ameliorated colitis and regulated immune cells; The macroscopic images of colon)( IDDF2021-ABS-0196 Figure 1C. Clostridium butyricum and its supernatant ameliorated colitis and regulated immune cells; The statistical analysis of colon length). The results of qPCR showed that the relative expression of pro-inflammatory cytokines TNF-α, IL-6, IL-1β, IL-17a were down-regulated after CBM and its supernatant treatment. And the relative expressions of anti-inflammatory cytokines TGF-β, IL-10, IL-4, IL-5, IL-13 were up-regulated (IDDF2021-ABS-0196 Figure1F. Clostridium butyricum and its supernatant ameliorated colitis and regulated immune cells; The amount of Th1 cells based on flow cytometry, and its statistical analysis). Flow cytometry showed that the amount of neutrophils, macrophage, Th1, Th17 cells were down-regulated which were the pro-inflammatory immune cells in intestinal mucosa lamina propria after CBM and its supernatant treatment (IDDF2021-ABS-0196 Figure1D. Clostridium butyricum and its supernatant ameliorated colitis and regulated immune cells; The amount of neutrophil based on flow cytometry, and its statistical analysis) (IDDF2021-ABS-0196 Figure1E. Clostridium butyricum and its supernatant ameliorated colitis and regulated immune cells; The amount of macrophage based on flow cytometry, and its statistical analysis) (IDDF2021-ABS-0196 Figure1F. Clostridium butyricum and its supernatant ameliorated colitis and regulated immune cells; The amount of Th1 cells based on flow cytometry, and its statistical analysis) (IDDF2021-ABS-0196 Figure1G. Clostridium butyricum and its supernatant ameliorated colitis and regulated immune cells; The amount of Th17 cells based on flow cytometry, and its statistical analysis) (IDDF2021-ABS-0196 Figure1H. Clostridium butyricum and its supernatant ameliorated colitis and regulated immune cells; The relative expression of inflammatory cytokines)

Conclusions

CBM can alleviate the symptoms and colon shortening of mice with colitis after pre-treatment with ABx, improve the expression of inflammatory factors, and might play an anti-inflammatory role by down-regulating the infiltration of inflammatory immune cells in intestinal mucosa lamina propria.