Abstract O157: H7, an enterohemorrhagic Escherichia coli (EHEC) strain, is a foodborne pathogen that can cause disease in livestock, poultry and humans (Homo sapiens). Magainins is a cationic peptide with antibacterial and antitumor properties, and its antibacterial activity against Gram-negative and Gram-positive bacteria has been reported in previous studies. The purpose of this study was to explore the effect of frog (Xenopus laevis) derived Magainin Ⅱ on intestinal inflammation and microflora in mice (Mus musculus) induced by EHEC O157: H7. Forty C57/BL6 male mice with similar weigh were randomly divided into 4 groups with 10 replicates in each group, CK group, O157 group, Mag Ⅱ group, O157+Mag Ⅱ group, respectively. All the groups received the same basal diet. The O157 group and O157+MagⅡgroup were orally administered 0.1 mL O157 bacterial solution with a concentration of 106 CFU/mL on 1, 5 and 9 d; CK group and MagⅡgroup were orally administered 0.1 mL PBS in the same time. After 1 h, MagⅡ and O157+MagⅡ group were intraperitoneally injected with 0.25 mL Magainin Ⅱ solution according to 5 mg/kg body weight, while CK and O157 group were intraperitoneally injected 0.25 mL sterilized saline. On the 10 d, mice were treatmented by euthanasia, and corresponding tissue samples were collected for testing. The results showed that Magainin Ⅱ treatment alone had no effect on the serum inflammatory cytokine concentration and the expression of jejunal inflammatory cytokines and barrier function related genes. EHEC O157: H7 treatment resulted in significant increase in the jejunum gene expression levels of interleukin-6 (IL-6), IL-1β, and tumor necrosis factor-α (TNF-α) and serum concentration of IL-1β, IL-6, TNF-α in mice (P<0.05). The serum IL-10 levels significantly decreased (P<0.05), as well as relative gene expression of jejunal tight junction proteins claudin-1, mucin-1 (MUC-1), and MUC-2 (P<0.05); Magainin Ⅱ treatment significantly improved EHEC O157: H7-induced relative gene expression of IL-1β, IL-6, TNF- α, claudin-1, MUC-1, MUC-2 in mouse jejunum and abnormal serum concentration of IL-1β, IL-6, IL-10 (P<0.05). EHEC O157: H7 infection could cause changes in intestinal microorganisms. The structure of intestinal microorganisms could be analyzed by 16S rDNA sequencing, and then the changes of intestinal microorganisms before and after Magainin Ⅱ intervention were analyzed. Sequencing results showed that at the genus level, compared with CK group, the relative abundance of Prevotella and Bacteroides in O157 group significantly increased (P<0.05). After MagaininⅡtreatment, the relative abundance of Bacteroides in O157+MagⅡ group decreased (P<0.05). The results showed that the abundance of Prevotella and Bacteroides related to the development of inflammation significantly increased after EHEC O157:H7 challenge, while the abundance of the unfavorable bacteria genus Bacteroides decreased significantly in the Magainin Ⅱ treatment group. This study showed that Magainin Ⅱ could maintain the intestinal barrier and alleviate the level of inflammation, thus playing a protective role in the intestinal injury induced by EHEC O157:H7 in mice, and the change of intestinal flora might be the related factor for Magainin Ⅱ to protect the intestinal injury. In conclusion, Magainin Ⅱ could be used as a potential drug for the treatment of intestinal diseases caused by EHEC O157:H7.
NONG Ke-Yi,FANG Xin,LIU You-Ming, et al. Effect of MagaininⅡ on Intestinal Barrier Injury and Inflammatory State Induced by EHEC O157:H7 in Mice (Mus musculus)[J]. 农业生物技术学报, 2023, 31(4): 812-823.
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