Mechanism of Zinc Relieves Inflammatory Response Mediated by Zinc Finger Protein A20 in Chicken Intestinal Epithelial Cells

GAO Jinxin, LI Changwu, ZHANG Beibei, GUO Shuangshuang, YUAN Jianmin, NIE Wei, GUO Yuming

doi:10.3969/j.issn.1006-267x.2019.05.032

2019 , Vol. 31 >Issue 5: 2254 - 2266

DOI: https://doi.org/10.3969/j.issn.1006-267x.2019.05.032

Molecular Nutrition

Mechanism of Zinc Relieves Inflammatory Response Mediated by Zinc Finger Protein A20 in Chicken Intestinal Epithelial Cells

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State Key Laboratory of Animal Nutrition, College of Animal Science and Technology, China Agricultural University, Beijing 100193, China

Received date: 2018-10-22

Online published: 2019-05-15

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Abstract

The research aimed to study the role of zinc and A20 gene in relieving lipopolysaccharide(LPS)-induced inflammatory response in chicken intestinal epithelial cells by infecting chicken intestinal epithelial cells with zinc finger protein A20 gene siRNA virus and A20 gene adenovirus. The results showed that LPS significantly increased the gene expression levels of pro-inflammatory cytokines such as interleukin (IL)-1β and IL-8 in chicken intestinal epithelial cells (P<0.05), and the optimal dose and time for LPS administration were 10 ng/mL and 6 h, respectively. Addition of 25 μmol/L zinc in vitro significantly alleviated LPS-induced higher gene expression levels and protein production of IL-8, IL-1β and tumor necrosis factor-α (TNF-α) in chicken intestinal epithelial cells, as well as the transcriptional activation of toll-like receptor 4 (TLR4) (P<0.05). After intestinal epithelial cells were infected with the siRNA virus of A20 gene, the protein expression of A20 was significantly down-regulated (P<0.05), the gene expression levels of IL-1, IL-8 and TNF-α and their protein production were significantly increased (P<0.05), and the LPS-induced up-regulated gene expression levels of IL-8 and IL-1β and protein production of IL-1β and TNF-α were also significantly enhanced (P<0.05). Compared with the control group, zinc addition was beneficial to the protein expression of A20 and nuclear factor-κB (NF-κB) p65, but inhibited protein expression of phospho-NF-κB p65. However, no effects of zinc on the down-regulated protein expression of proteins A20 and NF-κB p65 and up-regulated protein expression of phospho-NF-κB p65 and phospho-nuclear factor-κB inhibitory protein α (IκBα) in cytoplasm, and up-regulated protein expression of phospho-NF-κB p65 in nucleus under A20 gene silence were observed. Compared with the blank adenoviral group, infection with A20 gene adenovirus extremely significantly increased the gene expression level of A20 (P<0.01), extremely significantly decreased the gene expression levels of IL-1β and IL-8 (P<0.01), and the increasing gene expression levels of IL-1β, IL-8 and TLR4 induced by LPS were extremely significantly alleviated (P<0.01). Zinc addition significantly reduced the gene expression level of IL-1β and significantly increased the gene expression level of NF-κB p65(P<0.05). Therefore, A20-NF-κB p65 pathway plays an important role in the alleviation of LPS-induced inflammation by zinc.

Key words： broiler; primary intestine epithelial cells; zinc; A20; siRNA; adenovirus

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GAO Jinxin, LI Changwu, ZHANG Beibei, GUO Shuangshuang, YUAN Jianmin, NIE Wei, GUO Yuming . Mechanism of Zinc Relieves Inflammatory Response Mediated by Zinc Finger Protein A20 in Chicken Intestinal Epithelial Cells[J]. Chinese Journal of Animal Nutrition, 2019 , 31(5) : 2254 -2266 . DOI: 10.3969/j.issn.1006-267x.2019.05.032

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