Molecular Nutrition

Effects of Bacillus amyloliquefaciens SC06 and Escherichia coli K88 on Expression of Genes Related to Transporters, Tight Junctions, Apoptosis and Immunity in IPEC-1 Cells

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  • Key Laboratory of Molecular Animal Nutrition of Ministry of Education, Institute of Feed Science, College of Animal Science, Zhejiang University, Hangzhou 310058, China

Received date: 2018-10-22

  Online published: 2019-05-15

Abstract

This experiment was undertaken to evaluate the effects of Bacillus amyloliquefaciens SC06 (hereinafter referred to as SC06) and Escherichia coli K88 (hereinafter referred to as K88) on the expression of genes related to intestinal epithelial barrier function by material of IPEC-1 cells. The IPEC-1 cells were divided into four groups with different treatments: cells in group CK were not treated and used as blank control, while cells in group SC06 and group SC06+K88 were pretreated with DMEM/F12 medium containing 108 CFU/mL SC06 for 6 h, and then cells in group K88 and group SC06+K88 were treated with DMEM/F12 medium containing 108 CFU/mL K88 for 3 h. For the determination of dehydrogenase (LDH) activity, cells in group Trinton were treated with DMEM/F12 medium containing 1% Trinton X-100 and used as positive control. Cells in each group were cultured for 9 h. The results showed as follows: 1) compared with group CK, the relative expression levels of glucose transporter 2 (GLUT2) and small peptide transporter 1 (PepT1) genes were significantly decreased by K88 and SC06 alone treatment (P<0.05). SC06 alone treatment significantly increased the relative expression levels of alanine/serine/cysteine/threonine transporter 2 (ASCT2) and excitatory amino acid transporter 1 (EAAC1) genes (P<0.05). Compared with group, SC06 pretreatment significantly suppressed the higher LDH activity as well as the relative expression level of EAAC1 gene induced by K88 (P<0.05). 2) Compared with group CK, K88 alone treatment significantly upregulated the expression of zonula occludens protein-1 (ZO-1) and occludin genes (P<0.05), and significantly downregulated the expression of claudin-3, claudin-4 and mucin 1 (MUC1) genes (P<0.05). SC06 alone treatment significantly upregulated the expression of ZO-1 and claudin-4 genes (P<0.05). Both K88 and SC06 alone treatment significantly increased the expression of caspase-3, factor associated suicide (Fas) and B-cell lymphoma-2 (Bcl-2) genes (P<0.05), and significantly inhibited the expression of caspase-9 and Bcl-2 associated X protein (Bax) genes (P<0.05). Moreover, K88 alone treatment also significantly decreased the expression of caspase-8 gene (P<0.05). Compared with group K88, the increase expression of ZO-1, caspase-3 and Bcl-2 genes and the decrease expression of claudin-3, claudin-4, MUC1, caspase-9 and Bax genes induced by ETEC K88 were significantly prevented by SC06 pretreatment (P<0.05). 3) Compared with group CK, K88 alone treatment significantly upregulated the expression of tumor necrosis factor α (TNF-α), interleukin-6 (IL-6), interleukin-8 (IL-8) and transforming growth factor β (TGF-β) genes, and significantly upregulated the expression of nuclear factor κB-p50 (NF-κB-p50), tumor necrosis factor receptor-associated factor-6 (TRAF-6), myeloid differentiation factor 88 (MyD88), nucleotide binding oligomerization domain containing protein-1 (NOD-1) and Toll like receptor 4 (TLR4) genes (P<0.05). SC06 alone treatment significantly reduced the expression of IL-6, NOD-1 and Toll like receptor 6 (TLR6) genes (P<0.05), and significantly upregulated the expression of TGF-β and MyD88 (P<0.05). Compared with group K88, the increase expression of MyD88, NOD-1 and TLR4 induced by K88 were significantly prevented by SC06 pretreatment (P<0.05). In conclusion, K88 induces IPEC-1 cells inflammatory response and impairs integrity of epithelia, which is alleviated by SC06 to some extent.

Cite this article

XU Han, CAO Xuefang, LIU Rongrong, ZENG Zhonghua, TANG Li, LI Weifen . Effects of Bacillus amyloliquefaciens SC06 and Escherichia coli K88 on Expression of Genes Related to Transporters, Tight Junctions, Apoptosis and Immunity in IPEC-1 Cells[J]. Chinese Journal of Animal Nutrition, 2019 , 31(5) : 2267 -2277 . DOI: 10.3969/j.issn.1006-267x.2019.05.033

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