Inhibitory Effect of Clostridium butyricum on Inflammatory Reaction of Porcine Intestinal Epithelial Cells Induced by Enterotoxigenic Escherichia coli

YANG Hua; SHI Xingfen; GUI Guohong; LYU Wentao; LI Na; XIAO Yingping

doi:10.3969/j.issn.1006-267x.2019.12.035

2019 , Vol. 31 >Issue 12: 5688 - 5695

DOI: https://doi.org/10.3969/j.issn.1006-267x.2019.12.035

Inhibitory Effect of Clostridium butyricum on Inflammatory Reaction of Porcine Intestinal Epithelial Cells Induced by Enterotoxigenic Escherichia coli

YANG Hua ,
SHI Xingfen ,
GUI Guohong ,
LYU Wentao ,
LI Na ,
XIAO Yingping

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1. Institute of Quality and Standard for Agricultural Products, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, China;
2. Zhejiang Institute of Veterinary Drug and Feed Control, Hangzhou 310018, China

Received date: 2019-05-13

Online published: 2019-12-13

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Abstract

The aim of this study was to investigate the effects of Clostridium butyrium (CB) on the expression of the adhesion and enterotoxin genes in entertoxigenic Escherichia coli (ETEC) and the inflammation-associated factors in porcine intestinal epithelial cells (IPEC-J2 cells) challenged by ETEC in a co-culture model of CB, ETEC and IPEC-J2 cells. The experiment included 5 groups, they were control group, ETEC group, ETEC+CB 10⁶ group, ETEC+CB 10⁷ group, ETEC+CB 10⁸ group, respectively. In the ETEC group, IPEC-J2 cells were treated with 1×10⁸ CFU/mL ETEC, while IPEC-J2 cells in ETEC+CB 10⁶, ETEC+CB 10⁷ and ETEC+CB 10⁸ groups were treated with 1×10⁸ CFU/mL ETEC 1×10⁶, 1×10⁷, and 1×10⁸ CFU/mL CB, respectively. Neither ETEC nor CB was added in the IPEC-J2 cells culture solution of the control group. Cell morphology was analyzed with microscope after incubation at 37℃ for 2 h, and then cells were collected. The mRNA relative expression levels of adhesion gene FaeG and enterotoxigenic genes estA, estB in ETEC and pro-inflammatory factors interleukin (IL)-1β, IL-2, IL-6, IL-8 and anti-inflammatory factor IL-10 in IPEC-J2 cells were measured by quantitative real-time PCR (RT-qPCR). The results showed that CB could effectively inhibit the damage of IPEC-J2 cells induced by ETEC. Compared with ETEC group, different concentrations of CB treatment significantly reduced the expression of adhesion gene FaeG and enterotoxigenic genes estA, estB in ETEC (P<0.05), and the inhibitory effect was most obvious when the concentration of CB was 1×10⁸ CFU/mL. Compared with the control group, the mRNA relative expression levels of pro-inflammatory factors IL-1β, IL-2, IL-6 and IL-8 were significantly increased (P<0.05), whereas the mRNA relative expression level of anti-inflammatory factor IL-10 was significantly decreased in IPEC-J2 cells treated by ETEC (P<0.05). However, the mRNA relative expression levels of IL-1β, IL-2, IL-6, and IL-8 were significantly decreased (P<0.05), but the mRNA relative expression level of IL-10 was significantly increased in IPEC-J2 cells by different concentrations of CB treatment compared with ETEC group (P<0.05). The inhibitory effects of CB on the expression of IL-1β, IL-6 and IL-8 were most effective when the concentration was 1×10⁸ CFU/mL, and the inhibitory effects of CB on the expression of IL-2 was most effective when the concentration was 1×10⁷ CFU/mL. These results suggest that CB can prevent the damage of porcine intestinal epithelial cells induced by ETEC through the inhibitory effects of the expression of adhesion and enterotoxigenic genes in ETEC and inflammatory reaction in porcine intestinal epithelial cells.

Key words： Clostridium butyrium; entertoxigenic Escherichia coli; porcine intestinal epithelial cells; inflammatory reaction

Cite this article

YANG Hua , SHI Xingfen , GUI Guohong , LYU Wentao , LI Na , XIAO Yingping . Inhibitory Effect of Clostridium butyricum on Inflammatory Reaction of Porcine Intestinal Epithelial Cells Induced by Enterotoxigenic Escherichia coli[J]. Chinese Journal of Animal Nutrition, 2019 , 31(12) : 5688 -5695 . DOI: 10.3969/j.issn.1006-267x.2019.12.035

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