研究简报 Short Communications

营养、大肠杆菌和氨基酸对猪小肠上皮细胞抗菌肽和信号通路蛋白表达的影响

  • 任曼 ,
  • 宫碧霜 ,
  • 靳二辉 ,
  • 李升和 ,
  • 曾祥芳 ,
  • 谯仕彦
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  • 1. 安徽科技学院动物科学学院, 凤阳 233100;
    2. 中国农业大学动物科学学院, 北京 100193

收稿日期: 2015-12-25

  网络出版日期: 2016-05-24

基金资助

国家自然科学基金(31501968);安徽省高校自然科学研究项目(KJ2015A296);安徽省自然科学基金(1608085QC72);安徽科技学院高层次人才引进项目(ZRC2014453)

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

摘要

为了研究应激和氨基酸对上皮抗菌肽表达的作用和分子机制,试验选用猪小肠上皮细胞系IPEC-J2作为研究对象,饥饿和大肠杆菌感染分别作为营养应激和细菌应激,丙氨酸和异亮氨酸作为氨基酸处理,收集细胞mRNA,利用荧光定量PCR测定β防御素和相关信号通路蛋白表达水平。结果表明:与对照组(DMEM/F12培养基)相比,饥饿显著降低了小肠上皮细胞猪防御素2(pBD-2)、猪防御素3(pBD-3)和猪防御素EP2c(pEP2c)及沉默信息调节因子2同源蛋白1(Sirt1)、叉头转录因子1(FoxO1)和叉头转录因子4(FoxO4)的表达(P<0.05),大肠杆菌对β防御素表达无显著影响(P>0.05)。与对照组(饥饿培养基)相比,丙氨酸处理未显著影响pBD-2和pBD-3的表达(P>0.05),但显著提高了pEP2c表达水平(P<0.05);异亮氨酸处理使pBD-2、pBD-3和pEP2c表达水平显著升高(P<0.05)。与对照组相比,丙氨酸和异亮氨酸处理不同程度影响信号通路蛋白转录,丙氨酸处理显著提高了Sirt1和FoxO4的表达水平(P<0.05),异亮氨酸处理显著促进了Sirt1、FoxO1和FoxO4的表达(P<0.05)。由此得出,营养应激降低猪小肠上皮细胞中β防御素的表达,而氨基酸的补充可促进其表达,该调控作用可能与Sirt1和叉头转录因子(FoxO)信号通路蛋白有关。

本文引用格式

任曼 , 宫碧霜 , 靳二辉 , 李升和 , 曾祥芳 , 谯仕彦 . 营养、大肠杆菌和氨基酸对猪小肠上皮细胞抗菌肽和信号通路蛋白表达的影响[J]. 动物营养学报, 2016 , 28(5) : 1489 -1495 . DOI: 10.3969/j.issn.1006-267x.2016.05.025

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.

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