Objective To study the effect of flagellin on the adhesion of Clostridium butyricum to intestinal epithelial cells,and to provide an experimental basis for explaining the beneficial role of Clostridium butyricum as a probiotic.Methods The flagellin of Clostridium butyricum DSM 10702 was extracted and purified,and the protein was identified by mass spectrometry.The antiserum was obtained by immunizing mice with flagellin as an antigen,and the antiserum titer was determined by enzyme-linked immunosorbent assay(ELISA).Finally,human colon adenocarcinoma cell lines HT-29 and Caco-2 were used to construct an intestinal epithelial cell adhesion model,and flagellin was used to treat the cells and anti-flagellin and antiserum pretreatment bacteria.The t-test was used to compare the adhesion levels of the two treatment groups with that of the control group.Results The flagellin expressed by Clostridium butyricum DSM 10702 was encoded by FF104_14415 genes with a molecular weight of 29.56kDa,and the obtained antiserum titer was 1∶16 000.In the HT-29 cell adhesion model,compared with the control group,the adhesion of Clostridium butyricum in the flagellin pretreatment group and the anti-flagellin antiserum pretreatment group was significantly reduced,and the difference was statistically significant(t=2.870,6.135,P＜0.05).In the Caco-2 cell adhesion model,Clostridium butyricum adhesion was significantly reduced in the two treatment groups compared with the control group,and the difference was statistically significant(t=6.778,18.020,P＜0.05).Conclusion Flagellin contributes to Clostridium butyricum adhering to intestinal epithelial cells,and this study provides an experimental basis for further understanding of the adhesion mechanism of Clostridium butyricum.
Clostridium butyricumFlagellinAdhesionIntestinal epithelial cellAdhesion model
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