分子与细胞营养 MOLECULAR AND CELLULAR NUTRITION

基于转录组测序分析研究枸芪多糖对育肥猪肠黏膜免疫功能的作用机理

  • 霍金金 ,
  • 褚耀诚 ,
  • 金娜 ,
  • 李祯 ,
  • 郝壮 ,
  • 陈长增 ,
  • 李建东 ,
  • 刘凤华
展开
  • 1. 北京农学院动物科学技术学院, 北京 102206;
    2. 河北北方学院动物科技学院, 张家口 075000
霍金金(1992-),女,河北邯郸人,硕士研究生,从事中兽医药研究。E-mail:1075719601@qq.com

收稿日期: 2020-08-04

  网络出版日期: 2021-02-04

基金资助

北京市科技计划课题(D171100002117002)

Mechanism of Gouqi Polysaccharides on Intestinal Mucosal Immune Function of Finishing Pigs Based on Transcriptome Sequencing

  • HUO Jinjin ,
  • CHU Yaocheng ,
  • JIN Na ,
  • LI Zhen ,
  • HAO Zhuang ,
  • CHEN Changzeng ,
  • LI Jiandong ,
  • LIU Fenghua
Expand
  • 1. College of Animal Science and Technology, Beijing University of Agriculture, Beijing 102206, China;
    2. College of Animal Science and Technology, Hebei North University, Zhangjiakou 075000, China

Received date: 2020-08-04

  Online published: 2021-02-04

Supported by

 

摘要

本试验旨在研究枸芪多糖对育肥猪肠黏膜免疫调控的关键基因,阐明其对肠黏膜免疫功能的可能作用机理。选用80~90日龄的健康育肥母猪180头,随机分为2组,每组6个重复,每个重复15头猪。对照组饲喂基础饲粮,试验组在基础饲粮中添加0.1%的枸芪多糖,试验期90 d。采集育肥猪的空肠肠段,提取RNA后进行转录组测序分析差异免疫基因的表达。结果显示:1)与对照组相比,饲粮中添加枸芪多糖差异表达的免疫基因有75个,其中有26个上调基因和49个下调基因。2)基因本体(GO)富集分析显示差异表达基因主要参与免疫调节、免疫应答和应激反应,信号通路分析显示差异表达基因主要富集在以白细胞介素-17(IL-17)信号通路、NOD样受体信号通路为主的免疫信号通路。3)通过对肠黏膜差异表达的关键基因原癌基因Jun (JUN)和原癌基因Fos (FOS)的探究可见,与对照组相比,试验组白细胞介素-18(IL-18)、补体5(C5)、JUNFOS的基因表达受到抑制,Jun和Fos的蛋白表达显著降低(P<0.05)。综上所述,在饲粮中添加枸芪多糖可提高肥育猪的肠黏膜免疫功能。枸芪多糖通过调节JUNFOSIL-18等免疫相关基因的表达,影响IL-17和NOD样受体信号通路,以此来抑制炎症反应。

本文引用格式

霍金金 , 褚耀诚 , 金娜 , 李祯 , 郝壮 , 陈长增 , 李建东 , 刘凤华 . 基于转录组测序分析研究枸芪多糖对育肥猪肠黏膜免疫功能的作用机理[J]. 动物营养学报, 2021 , 33(2) : 1100 -1110 . DOI: 10.3969/j.issn.1006-267x.2021.02.050

Abstract

This experiment aimed to study the key genes of Gouqi polysaccharide on intestinal mucosal immune regulation of finishing pigs and clarify functional mechanism of Gouqi polysaccharide on intestinal mucosal immune. One hundred and eighty healthy female finishing pigs with 80 to 90 days of age were randomly divided into 2 groups with 6 replicates per group and 15 pigs per replicate. Pigs in control group were fed a basal diet, and the others in experimental group were fed the basal diet supplemented with 0.1% Gouqi polysaccharide. The experiment lasted for 90 days. RNA was extracted from jejunum of finishing pigs and then differential immune genes expression were analyzed by transcriptome sequencing. The results showed as follows: 1) compared with control group, there are 75 differentially expressed immune genes by dietary Gouqi polysaccharide including 26 up-regulated genes and 49 down-regulated genes. 2) Gene ontology (GO) enrichment analysis showed that differentially expressed genes were mainly involved in immune regulation, immune response and stress response. Signaling pathway analysis showed that differentially expressed genes were mainly enriched in immune signaling pathway mainly composed of interleukin-17 (IL-17) signaling pathway and nucleotide binding oligomerization domain (NOD) like receptors signaling pathway. 3) Exploring key genes of differential expression in intestinal mucosa JUN and FOS showed that, compared with control group, genes expression of interleukin-18 (IL-18), complement 5 (C5), JUN and FOS in experimental group was inhabited, and protein expression of Jun and Fos were significantly decreased (P<0.05). In conclusion, dietary Gouqi polysaccharide can enhance intestinal mucosal immune function of finishing pigs. Gouqi polysaccharide affect IL-17 and NOD-like receptor signaling pathways by regulating expressions of immune-related genes such as JUN, FOS and IL-18, thereby inhibiting inflammation.

参考文献

[1] VANCAMELBEKE M,VERMEIRE S.The intestinal barrier:A fundamental role in health and disease[J].Expert Review of Gastroenterology & Hepatology,2017,11(9):821-834.  
[2] MORETÓ M,PÉREZ-BOSQUE A.Dietary plasma proteins,the intestinal immune system,and the barrier functions of the intestinal mucosa[J].Journal of Animal Science,2009,87(Suppl.14):E92-E100.
[3] 高倩,刘凤华,史雅然,等.一种基于酶标仪检测天然植物复方多糖中多糖含量的方法[J].动物营养学报,2019,31(2):922-929. GAO Q,LIU F H,SHI Y R.et al. Determination method of polysaccharide content in natural plant polysaccharide compound based on microplate reader[J].Chinese Journal of Animal Nutrition,2019,31(2):922-929.(in Chinese)
[4] WANG Y X,CHEN Y,DU H X,et al.Comparison of the anti-duck hepatitis a virus activities of phosphorylated and sulfated Astragalus polysaccharides[J].Experimental Biology and Medicine,2017,242(3):344-353.  
[5] FU J,WANG Z H,HUANG L F,et al.Review of the botanical characteristics,phytochemistry,and pharmacology of Astragalus membranaceus (Huangqi)[J].Phytotherapy Research,2014,28(9):1275-1283.  
[6] LI X X,QU L,DONG Y Z,et al.A review of recent research progress on the Astragalus genus[J].Molecules,2014,19(11):18850-18880.  
[7] 娜日苏,孔维雪.枸杞多糖的纯化与生理功能的研究进展[J].中药材,2019(3):704-707. NA R S,KONG W X.Research progress on purification and physiological function of Lycium barbarum polysaccharide[J].Journal of Chinese Medicinal Materials,2019(3):704-707.(in Chinese)
[8] DESOUSA L M S,NASCIMENTO W M,COSTA D D A F,et al.Evaluate the action of the aqueous extract of the medicinal plant Goji Berry (Lyciumbarbarum) on biochemical and hematological parameters in Wistar rats[J].IOSR Journal of Pharmacy and Biological Sciences,2016,11(5):48-52.
[9] 孙文丽,SHAHRAJABIAN M H,程奇.枸杞化学成分及药用价值国外研究现状[J].中医药信息,2020,37(3):116-120. SUN W L,SHAHRAJABIAN M H,CHEN Q.Overseas research status of chemical components and phamaceutical value of Lycium barbarum[J].Information on Traditional Chinese Medicine,2020,37(3):116-120.(in Chinese)
[10] 陶艳华,金福源,陆晓健,等.黄芪多糖对仔猪免疫力以及生长性能的影响[J].养殖与饲料,2019(1):40-42. TAO Y H,JIN F Y,LU X J,et al.Effects of Astragalus polysaccharide on immunity and growth performance of piglets[J]Breeding and Feed,2019(1):40-42.(in Chinese)
[11] 王思月.枸杞多糖对肉鸡肠道微生物区系、免疫功能及相关基因表达的影响[D].硕士学位论文.长春:吉林农业大学,2019. WANG S Y.Effects of Lycium barbarum polysaccharides on intestinal microflora,immune function and expression of related genes in broilers[D].Master's Thesis.Changchun:Jilin Agricultural University,2019.(in Chinese)
[12] 孙晓雨,崔子寅,张明亮,等.枸杞多糖和茯苓多糖对免疫抑制小鼠免疫增强及对肠道黏膜的免疫调节作用[J].中国兽医学报,2015,35(3):450-455. SUN X Y,CUI Z Y,ZHANG M L,et al.Effects of Lycium barbarum polysaccharides and pachymaran on immune function and intestinal mucosal system in immunosuppressive mice[J].Chinese Journal of Veterinary Science,2015,35(3):450-455.(in Chinese)
[13] 张乐萃,王金宝,孙月平,等.复方中药多糖对鸡免疫器官形态学的影响[J].中国兽医科学,1998,28(8):26-28. ZHANG L C,WANG J B,SUN Y P,et al.Effects of compound Chinese medicine polysaccharide on morphology of chicken immune organs[J].Chinese Veterinary Science,1998,28(8):26-28.(in Chinese)
[14] 刘保光,吴华,徐利纳,等.黄芪多糖的药理学功效及在兽医临床的应用[J].广东饲料,2010,19(6):30-32. LIU B G,WU H,XU L N,et al.The pharmacological effect of Astragalus polysaccharide and its clinical application in veterinary medicine[J].Guangdong Feed,2010,19(6):30-32.(in Chinese)
[15] 孙永涛.黄芪多糖对鸡新城疫疫苗免疫后T淋巴细胞的影响[J].当代畜牧,2018(8):63-64. SUN Y T,Effects of Astragalus polysaccharides on T lymphocytes of chickens immunized with Newcastle disease vaccine.[J].Contemporary Animal Husbandry,2018(8):63-64.(in Chinese)
[16] CHEN Z S,TAN B K H,CHAN S H.Activation of T lymphocytes by polysaccharide-protein complex from Lycium barbarum L.[J].International Immunopharmacology,2008,8(12):1663-1671.  
[17] 金娜,邹闻书,高倩,等.枸芪多糖对育肥猪生长性能、胴体性状和肠道组织形态的影响[J].北京农学院学报,2020,35(1):55-59. JIN N,ZOU W S,GAO Q,et al.Effects of GouQi polysaccharide on growth performance, carcass traits and intestinal tissue morphology of growing-finishing pigs[J].Journal of Beijing University of Agriculture,2020,35(1):55-59.(in Chinese)
[18] 金娜,邹闻书,高倩,等.枸芪多糖对育肥猪肠道菌群多样性及组成的影响[J].动物营养学报,2019,31(9):4033-4043. JIN N,ZOU W S,GAO Q,et al.Effects of Gou-Qi polysaccharide on intestinal microflora diversity and composition of growing-finishing pigs[J].Chinese Journal of Animal Nutrition,2019,31(9):4033-4043.(in Chinese)
[19] ABREU-MARTIN M T,TARGAN S R.Regulation of immune responses of the intestinal mucosa[J].Critical Reviews in Immunology,1996,16(3):227-309.
[20] JING L,HOU Y,WU H,et al.Transcriptome analysis of mRNA and miRNA in skeletal muscle indicates an important network for differential residual feed intake in pigs[J].Scientific Reports,2015,5:11953.
[21] ZHAO Y X,HOU Y,LIU F,et al.Transcriptome analysis reveals that vitamin a metabolism in the liver affects feed efficiency in pigs[J].Genes,Genomes,Genetics,2016,6(11):3615-3624.
[22] VIERBUCHEN T,LING E,COWLEY C J,et al.AP-1 transcription factors and the BAF complex mediate signal-dependent enhancer selection[J].Molecular Cell,2017,68(6):1067-1082.e12.  
[23] MAKINO H,SEKI S,YAHARA Y,et al.A selective inhibition of c-Fos/activator protein-1 as a potential therapeutic target for intervertebral disc degeneration and associated pain[J].Scientific Reports,2017,7(1):16983.
[24] HIROI N,NESTLER E J.Nuclear memory:gene transcription and behavior[J].Advances in Pharmacology,1997,42:1037-1041.
[25] OKAMURA H,NAGATA K,KOMASTU T,et al.A novel costimulatory factor for gamma interferon induction found in the livers of mice causes endotoxic shock[J].Infection and Immunity,1995,63(10):3966-3972.  
[26] 胡厚祥,李保胜,刘利,等.过敏毒素C5a通过Calpain途径致VEC凋亡损伤[J].免疫学杂志,2009,25(4):424-427. HU H X,LI B S,LIU L,et al.rhC5a induces apoptosis by stimulating and regulating the expression of calpain in endothelial cells[J].Immunological Journal,2009,25(4):424-427.(in Chinese)
文章导航

/