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当前位置: 首页 > 文章 > 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 动物营养学报 2016,28 (5) 1489-1495
Position: Home > Articles > Effects of Nutrition,Escherichia coli and Amino Acids on Expression of Antimicrobial Peptide and Signaling Pathway Protein in Porcine Intestinal Epithelial Cells Chinese Journal of Animal Nutrition 2016,28 (5) 1489-1495

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

作  者：
任曼;宫碧霜;靳二辉;李升和;曾祥芳;谯仕彦
单  位：
安徽科技学院动物科学学院;中国农业大学动物科学学院
关键词：
猪小肠上皮细胞;抗菌肽;防御素;信号通路;Sirt1;Fox O
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(p BD-2)、猪防御素3(p BD-3)和猪防御素EP2c(p EP2c)及沉默信息调节因子2同源蛋白1(Sirt1)、叉头转录因子1(Fox O1)和叉头转录因子4(Fox O4)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p BD-2和p BD-3的表达(P>0.05),但显著提高了p EP2c表达水平(P<0.05);异亮氨酸处理使p BD-2、p BD-3和p EP2c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Fox O4的表达水平(P<0.05),异亮氨酸处理显著促进了Sirt1、Fox O1和Fox O4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(Fox O)信号通路蛋白有关。
译  名：
Effects of Nutrition,Escherichia coli and Amino Acids on Expression of Antimicrobial Peptide and Signaling Pathway Protein in Porcine Intestinal Epithelial Cells
作  者：
REN Man;GONG Bishuang;JIN Erhui;LI Shenghe;ZENG Xiangfang;QIAO Shiyan;College of Animal Science,Anhui Science and Technology University;College of Animal Science and Technology,China Agricultural University;
关键词：
porcine intestinal epithelial cell;;antimicrobial peptide;;defensin;;signaling pathway;;Sirt1;;FoxO
摘  要：
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 realtime quantitative PCR.From results,we found compared with the control group(with DM EM/F12 medium),starvation treatment significantly decreased the expression levels of porcine β-defensin 2(p BD-2),porcine β-defensin 2(p BD-3),porcine β-defensin EP2c(p EP2c),silent mating type information regulation 2 homolog1(Sirt1),forkhead transcription factor 1(Fox O1) and forkhead transcription factor 4(Fox O4)(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 p EP2 c expression level(P<0.05),but did not significantly change the expression of p BD-2 and p BD-3,and the expression levels of p BD-2,p BD-3 and p EP2 c 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 Fox O4 in alanine group were significantly increased and isoleucine treatment significantly increased the expression of Sirt1,Fox O1 and Fox O4(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(Fox O)signaling pathway protein.

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