Effects of Nutrition, Escherichia coli and Amino Acids on Expression of Antimicrobial Peptide and Signaling Pathway Protein in Porcine Intestinal Epithelial Cells

REN Man; GONG Bishuang; JIN Erhui; LI Shenghe; ZENG Xiangfang; QIAO Shiyan

doi:10.3969/j.issn.1006-267x.2016.05.025

2016 , Vol. 28 >Issue 5: 1489 - 1495

DOI: https://doi.org/10.3969/j.issn.1006-267x.2016.05.025

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Effects of Nutrition, Escherichia coli and Amino Acids on Expression of Antimicrobial Peptide and Signaling Pathway Protein in Porcine Intestinal Epithelial Cells

REN Man ,
GONG Bishuang ,
JIN Erhui ,
LI Shenghe ,
ZENG Xiangfang ,
QIAO Shiyan

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1. College of Animal Science, Anhui Science and Technology University, Fengyang 233100, China;
2. College of Animal Science and Technology, China Agricultural University, Beijing 100193, China

Received date: 2015-12-25

Online published: 2016-05-24

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Abstract

To investigate the effects and molecular mechanism of stress and amino acids on epithelial antimicrobial peptide, porcine intestinal epithelial cell line IPEC-J2 was treated with starvation (as nutritional stress), Escherichia coli (as bacterial stress), alanine or isoleucine (both as amino acid treatments), and cell mRNA was collected. Then the expression levels of β-defensins and signaling pathway protein were tested using real-time quantitative PCR. From results, we found compared with the control group (with DMEM/F12 medium), starvation treatment significantly decreased the expression levels of porcine β-defensin 2(pBD-2), porcine β-defensin 2(pBD-3), porcine β-defensin EP2c (pEP2c), silent mating type information regulation 2 homolog 1(Sirt1), forkhead transcription factor 1(FoxO1) and forkhead transcription factor 4(FoxO4) (P<0.05), but Escherichia coli treatment had no significant effects on β-defensin expression (P>0.05). Compared with the control group (with starvation medium), alanine treatment significantly increased pEP2c expression level (P<0.05), but did not significantly change the expression of pBD-2 and pBD-3, and the expression levels of pBD-2, pBD-3 and pEP2c in isoleucine group were significantly increased (P<0.05). Signaling pathway protein transcription was different affected by alanine and isoleucine treatments. Compared with the control group, the expression levels of Sirt1 and FoxO4 in alanine group were significantly increased and isoleucine treatment significantly increased the expression of Sirt1, FoxO1 and FoxO4 (P<0.05). In conclusion, nutritional stress can increase β-defensin expression in intestinal epithelial cells, and amino acid supplementation can promote its expression. The mechanism of defensin regulation is related to Sirt1 and forkhead transcription factor (FoxO) signaling pathway protein.

Key words： porcine intestinal epithelial cell; antimicrobial peptide; defensin; signaling pathway; Sirt1; FoxO

Cite this article

REN Man , GONG Bishuang , JIN Erhui , LI Shenghe , ZENG Xiangfang , QIAO Shiyan . Effects of Nutrition, Escherichia coli and Amino Acids on Expression of Antimicrobial Peptide and Signaling Pathway Protein in Porcine Intestinal Epithelial Cells[J]. Chinese Journal of Animal Nutrition, 2016 , 28(5) : 1489 -1495 . DOI: 10.3969/j.issn.1006-267x.2016.05.025

References

[1] ZHU Y G,JOHNSON T A,SU J Q,et al.Diverse and abundant antibiotic resistance genes in Chinese swine farms[J].Proceeding of the National Academy of Science United States of America,2013,110(9):3435-3440.

[2] LOOFT T,JOHNSON T A,ALLEN H K,et al.In-feed antibiotic effects on the swine intestinal microbiome[J].Proceeding of the National Academy of Science United States of America,2012,109(5):1691-1696.

[3] VELDHUIZEN E J A,RIJNDERS M,CLAASSEN E A,et al.Porcine β-defensin 2 displays broad antimicrobial activity against pathogenic intestinal bacteria[J].Molecular Immunology,2008,45(2):386-394.

[4] GALLO R L,HOOPER L V.Epithelial antimicrobial defence of the skin and intestine[J].Nature Reviews Immunology,2012,12(7):503-516.

[5] 任曼,霍应峰,杨凤娟,等.仔猪断奶前后肠道形态和相关免疫蛋白基因表达的变化[J].动物营养学报,2014,26(3):614-619.
[6] DI NARDO A,BRAFF M H,TAYLOR K R,et al.Cathelicidin antimicrobial peptides block dendritic cell TLR4 activation and allergic contact sensitization[J].Journal of Immunology,2007,178(3):1829-1834.

[7] ZENG X F,SUNKARA L T,JIANG W Y,et al.Induction of porcine host defense peptide gene expression by short-chain fatty acids and their analogs[J].PLoS One,2013,8(8):E72922.
[8] REN M,ZHANG S H,ZENG X F,et al.Branched-chain amino acids are beneficial to maintain growth performance and intestinal immune-related function in weaned piglets fed protein restricted diet[J].Asian Australasian Journal of Animal Sciences,2015,28(12):1742-1750.

[9] ZASLOFF M.Antimicrobial peptides of multicellular organisms[J].Nature,2002,415(6870):389-395.

[10] SANTAOLALLA R,FUKATA M,ABREU M T.Innate immunity in the small intestine[J].Current Opinion in Gastroenterology,2011,27(2):125-131.

[11] MAO X B,QI S,YU B,et al.Dietary L-arginine supplementation enhances porcine β-defensins gene expression in some tissues of weaned pigs[J].Livestock Science,2012,148(1/2):103-108.
[12] MAO X B,QI S,YU B,et al.Zn²⁺ and L-isoleucine induce the expressions of porcine β-defensins in IPEC-J2 cells[J].Molecular Biology Reports,2013,40(2):1547-1552.

[13] MUTURI E J,KIM C H,ALTO B W,et al.Larval environmental stress alters Aedes aegypti competence for Sindbis virus[J].Tropical Medicine & International Health,2011,16(8):955-964.

[14] BECKER T,LOCH G,BEYER M,et al.FOXO-dependent regulation of innate immune homeostasis[J].Nature,2010,463(7279):369-373.

[15] AKODA K,VAN DEN BOSSCHE P,MARCOTTY T,et al.Nutritional stress affects the tsetse fly's immune gene expression[J].Medical and Veterinary Entomology,2009,23(3):195-201.

[16] FEHLBAUM P,RAO M,ZASLOFF M,et al.An essential amino acid induces epithelial β-defensin expression[J].Proceeding of the National Academy of Science United States of America,2000,97(23):12723-12728.

[17] SHERMAN H,CHAPNIK N,FROY O.Albumin and amino acids upregulate the expression of human beta-defensin 1[J].Molecular Immunology,2006,43(10):1617-1623.

[18] KONNO Y,ASHIDA T,INABA Y,et al.Isoleucine,an essential amino acid,induces the expression of human β defensin 2 through the activation of the G-protein coupled receptor-ERK pathway in the intestinal epithelia[J].Food and Nutrition Sciences,2012,3(4):548-555.

[19] WEHKAMP J,HARDER J,WEHKAMP K,et al.NF-κB- and AP-1-mediated induction of human beta defensin-2 in intestinal epithelial cells by Escherichia coli Nissle 1917:a novel effect of a probiotic bacterium[J].Infection and Immunity,2004,72(10):5750-5758.

[20] YONEDA J,ANDOU A,TAKEHANA K.Regulatory roles of amino acids in immune response[J].Current Rheumatology Reviews,2009,5(4):252-258.

[21] DE SIMONED R,VISSICCHIO F,MINGARELLI C,et al.Branched-chain amino acids influence the immune properties of microglial cells and their responsiveness to pro-inflammatory signals[J].Biochimica et Biophysica Acta:Molecular Basis of Disease,2013,1832(5):650-659.

[22] TANNO M,SAKAMOTO J,MIURA T,et al.Nucleocytoplasmic shuttling of the NAD⁺-dependent histone deacetylase SIRT1[J].Journal of Biological Chemistry,2006,282(9):6823-6832.

[23] NARALA S R,ALLSOPP R C,WELLS T B,et al.SIRT1 acts as a nutrient-sensitive growth suppressor and its loss is associated with increased AMPK and telomerase activity[J].Molecular Biology of the Cell,2008,19(3):1210-1219.
[24] D'ANTONA G,RAGNI M,CARDILE A,et al.Branched-chain amino acid supplementation promotes survival and supports cardiac and skeletal muscle mitochondrial biogenesis in middle-aged mice[J].Cell Metabolism,2010,12(4):362-372.

[25] JONSSON H,PENG S L.Forkhead transcription factors in immunology[J].Cellular and Molecular Life Sciences,2005,62(4):397-409.

[26] NEMOTO S,FERGUSSON M M,FINKEL T.Nutrient availability regulates SIRT1 through a forkhead-dependent pathway[J].Science,2004,306(5704):2105-2108.

[27] LEE S I,MIN K S,BAE W J,et al.Role of SIRT1 in heat stress- and lipopolysaccharide-induced immune and defense gene expression in human dental pulp cells[J].Journal of Endodontics,2011,37(11):1525-1530.

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