Sanguinarine Inhibits Inflammatory Response of Piglet Small Intestinal Mucosal Epithelial Cells Induced by Lipopolysaccharide through Nuclear Factor-&kappa;B Signaling Pathway

GAO Huan; JIANG Xiaoxu; YANG Yuting; RAN Jinming; SHEN Liyan; JIANG Hengxin; LI Xueyan; LIN Qiuye; JIANG Qingwen; CAO Zhenhui; PAN Hongbin

doi:10.3969/j.issn.1006-267x.2021.03.048

2021 , Vol. 33 >Issue 3: 1675 - 1686

DOI: https://doi.org/10.3969/j.issn.1006-267x.2021.03.048

Sanguinarine Inhibits Inflammatory Response of Piglet Small Intestinal Mucosal Epithelial Cells Induced by Lipopolysaccharide through Nuclear Factor-κB Signaling Pathway

GAO Huan ,
JIANG Xiaoxu ,
YANG Yuting ,
RAN Jinming ,
SHEN Liyan ,
JIANG Hengxin ,
LI Xueyan ,
LIN Qiuye ,
JIANG Qingwen ,
CAO Zhenhui ,
PAN Hongbin

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1. Yunnan Key Laboratory of Animal Nutrition and Feed, College of Animal Science and Technology, Yunnan Agricultural University, Kunming 650201, China;
2. Dazhou Vocational and Technical College, Dazhou 635000, China;
3. Institute of Spices Yunnan Agricultural University, Kunming 650201, China

Received date: 2020-08-22

Online published: 2021-03-18

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Abstract

This study was to elucidate the anti-inflammatory regulate mechanism of sanguinarine (SG) by nuclear factor-κB (NF-κB) signaling pathway in piglet small intestinal mucosal epithelial cells (IPEC-J2 cells). In this study, we investigated the appropriate concentration and time for SG to promote the growth of IPEC-J2 cells and the appropriate concentration and time for LPS to establish an inflammatory model of IPEC-J2 cells. The expression of the key genes and protein expression and product content in NF-κB signal pathway among the control group (no added), inflammation model cell group (LPS group, 5.0 μg/mL LPS treatment for 1 h) and appropriate concentration of SG+LPS-induced inflammation model cell group (SG+LPS group, 5.0 μg/mL LPS treatment for 1 h and 0.500 μg/mL SG treatment for 24 h) were detected. The results showed as follows: 1) the appropriate concentration and time for SG to promote the growth of IPEC-J2 cells were 0.500 μg/mL and 24 h, respectively, and the appropriate concentration and time for the construction of IPEC-J2 cell inflammatory model of LPS were 5.0 μg/mL and 1 h, respectively. 2) Compared with the control group, the mRNA relative expression levels of interleukin-6 (IL-6), interleukin-8 (IL-8), nuclear factor-κB subunit 1 (NF-κB1A), nuclear factor-κB2 (NF-κB2), tumor necrosis factor-α inducible protein 3 (TNFAIP3), nuclear factor-κB kinase inhibitors ε (IKKε), nuclear factor-κB1 (NF-κB1) and RelB in the LPS group were significantly increased (P<0.01); the contents of IL-6, IL-8 and TNF-α in cell supernatant were significantly increased (P<0.05 or P<0.01); the protein expression levels of nuclear factor-κB inhibitor protein α (IκBα) and p65 in cells total protein were significantly decreased (P<0.05), the nucleoprotein p65 protein expression level was significantly increased (P<0.01), and the cytoplasmic protein p65 protein expression level was significantly decreased (P<0.01). 3) Compared with the LPS group, the mRNA relative expression levels of IL-6, IL-8, NF-κB1A, NF-κB2, TNFAIP3, IKKε and NF-κB1 in the SG+LPS group were significantly decreased (P<0.05 or P<0.01); the contents of IL-6 and IL-8 in cell supernatant were significantly decreased (P<0.05 or P<0.01); the cells total protein IκBα protein expression level was significantly increased (P<0.01), the nucleoprotein p65 protein expression level was significantly decreased (P<0.01), and the cytoplasmic protein p65 protein expression level was significantly increased (P<0.01). In conclusion, the SG can reduce the inflammation of IPEC-J2 cells by inhibiting the related gene activation of NF-κB signaling pathway and p65 nuclear transfer.

Key words： sanguinarine; IPEC-J2 cells; NF-κB signaling pathway; anti-inflammatory

Cite this article

GAO Huan , JIANG Xiaoxu , YANG Yuting , RAN Jinming , SHEN Liyan , JIANG Hengxin , LI Xueyan , LIN Qiuye , JIANG Qingwen , CAO Zhenhui , PAN Hongbin . Sanguinarine Inhibits Inflammatory Response of Piglet Small Intestinal Mucosal Epithelial Cells Induced by Lipopolysaccharide through Nuclear Factor-κB Signaling Pathway[J]. Chinese Journal of Animal Nutrition, 2021 , 33(3) : 1675 -1686 . DOI: 10.3969/j.issn.1006-267x.2021.03.048

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