丁酸梭菌对2种遗传背景仔猪肠道屏障功能的影响及其分子机制

李海花; 李玉鹏; 王柳懿; 王文杰; 乔家运

doi:10.3969/j.issn.1006-267x.2019.10.029

动物营养学报 >

2019 , Vol. 31 >Issue 10: 4647 - 4658

DOI: https://doi.org/10.3969/j.issn.1006-267x.2019.10.029

分子营养 Molecular Nutrition

丁酸梭菌对2种遗传背景仔猪肠道屏障功能的影响及其分子机制

李海花 ,
李玉鹏 ,
王柳懿 ,
王文杰 ,
乔家运

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1. 天津农学院动物科学与动物医学学院, 天津 300384;
2. 天津市畜牧兽医研究所, 天津 300381;
3. 天津师范大学生命科学学院, 天津 300387

李海花(1981-),女,河南周口人,副研究员,博士,研究方向为动物营养与免疫。E-mail:lihaihuaok@126.com

收稿日期: 2019-03-05

网络出版日期: 2019-10-17

基金资助

国家自然科学基金项目（31702147）；天津市农业科学院青年科研人员创新研究与实验项目（编号：2018008）；天津市""131"创新型人才团队（20180338）

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Effects of Clostridium butyricum on Intestinal Barrier Function of Piglets with Two Genetic Backgrounds and Its Molecular Mechanism

LI Haihua ,
LI Yupeng ,
WANG Liuyi ,
WANG Wenjie ,
QIAO Jiayun

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1. College of Animal Science and Animal Medicine, Tianjin Agricultural University, Tianjin 300384, China;
2. Tianjin Institute of Animal Husbandry and Veterinary, Tianjin 300381, China;
3. Tianjin Normal University, Tianjin 300387, China

Received date: 2019-03-05

Online published: 2019-10-17

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摘要

本试验旨在研究丁酸梭菌对2种遗传背景断奶仔猪健康状况、肠道发育、肠道屏障功能、血清细胞因子含量和黏膜免疫关键蛋白表达的影响。试验采用2×2因子设计，选择（21±2）日龄、体重相近的"东北民猪×巴克夏"二元杂交黑猪12头，随机分成2个组（对照组和试验组）；另选择（21±2）日龄、体重相近的"杜×长×大"三元杂交白猪12头，随机分成2个组（对照组和试验组）；每个组均6个重复，单栏饲养。对照组均饲喂基础饲粮，试验组均饲喂基础饲粮+5×10¹¹ CFU/kg丁酸梭菌制剂。试验预试期3 d，正试期14 d。结果表明：1）与相应的对照组相比，2个试验组仔猪血小板数目均显著增加（P<0.05），空肠和回肠的绒毛高度/隐窝深度均显著提高（P<0.05），血清中D-乳酸含量均显著降低（P<0.05）；二元杂交试验组仔猪空肠封闭蛋白1（claudin-1）和回肠claudin-1、紧密连接蛋白1（ZO-1）、闭合蛋白（occludin）mRNA相对表达水平均显著提高（P<0.05），三元杂交试验组仔猪空肠ZO-1和occludin mRNA相对表达水平均显著提高（P<0.05）；二元杂交试验组仔猪空肠黏膜中Toll样受体2（TLR2）、Toll样受体4（TLR4）、髓样分化因子88（MyD88）和回肠黏膜中TLR2、TLR4、B细胞淋巴瘤蛋白3（Bcl3）mRNA相对表达水平均显著提高（P<0.05），三元杂交试验组仔猪空肠黏膜中TLR2和回肠黏膜中TLR2、TLR4、MyD88、Toll作用蛋白（Tollip）、Bcl3 mRNA相对表达水平均显著提高（P<0.05）。2）二元杂交试验组仔猪空肠claudin-1、TLR4和MyD88和回肠claudin-1、ZO-1、occludin mRNA相对表达水平显著高于三元杂交试验组仔猪（P<0.05），但是二元杂交试验组仔猪回肠黏膜中TLR2、TLR4、Tollip和Bcl3 mRNA相对表达水平显著低于三元杂交试验组仔猪（P<0.05）。综上所述，丁酸梭菌能够促进仔猪肠道发育，激活2种遗传背景仔猪肠黏膜TLR2/4信号通路，并促进其通路关键负反馈子的表达，提高紧密连接蛋白的表达，降低肠黏膜通透性，提高肠道屏障功能，并且对2种遗传背景仔猪肠道屏障功能的调节存在差异。

关键词： 丁酸梭菌; 二元杂交; 三元杂交; 断奶仔猪; Toll样受体2; Toll样受体4

本文引用格式

李海花 , 李玉鹏 , 王柳懿 , 王文杰 , 乔家运 . 丁酸梭菌对2种遗传背景仔猪肠道屏障功能的影响及其分子机制[J]. 动物营养学报, 2019 , 31(10) : 4647 -4658 . DOI: 10.3969/j.issn.1006-267x.2019.10.029

Abstract

This study was aimed to investigate the effects of Clostridium butyricum on the health status, intestinal development, intestinal barrier function, serum cytokine contents and mucosal immunity key protein expression of weaned piglets with two genetic backgrounds. A 2×2 factors design was used in this experiment, 12 "Northeastern indigenous×Berkshire" black piglets with similar body weight and (21±2) days of age were selected and divided into 2 groups (control group and experimental group), randomly, and 12 "Duroc×Landrace×Yorkshire" white piglets with similar body weight and (21±2) days of age were selected and divided into 2 groups (control group and experimental group), randomly. Each group was divided into 6 replicates and each pig was fed in single column. Weaned piglets in control groups were fed basal diets, and those in experimental groups were fed the basal diets supplemented with 5×10¹¹ CFU/kg Clostridium butyricum preparation for 14 days after 3 days adaption. The results showed as follows:1) compared with the each control group, the platelet counts of piglets in the two experimental groups were both increased significantly (P<0.05), the villus length to crypt depth ratios in both jejunum and ileum of the two kinds of piglets were increased significantly (P<0.05), the serum D-lactic contents of the two kinds of weaned piglets were decreased significantly (P<0.05), the claudin-1 mRNA relative expression level in jejunum, and the mRNA relative expression levels of claudin-1, zonula occludens-1 (ZO-1) and occludin in ileum of piglets in binary hybridization experimental group were all increased significantly (P<0.05), the mRNA relative expression levels of ZO-1 and occludin in jejnum of piglets in ternary hybridization experimental group were both increased significantly (P<0.05), the mRNA relative expression levels of Toll-like receptor 2 (TLR2), Toll-like receptor 4 (TLR4) and myeloid differentiation factor 88 (MyD88) in jejunum mucosa and TLR2, TLR4 and B-cell lymphoma 3 protein (Bcl3) in ileum mucosa of piglets in binary hybridization experimental group were increased significantly (P<0.05), and the mRNA relative expression levels of TLR2 in jejunum mucosa and TLR2, TLR4, MyD88, Toll-interacting protein (Tollip) and Bcl3 in ileum mucosa of piglets in ternary hybridization experimental group were increased significantly (P<0.05). 2) Compared with the ternary hybridization experimental group, the mRNA relative expression levels of claudin-1, TLR4 and MyD88 in jejunum and claudin-1, ZO-1 and occludin in ileum of piglets in binary hybridization experimental group were increased significantly (P<0.05), but the mRNA relative expression levels of TLR2, TLR4, Tollip and Bcl3 in ileum mucosa of piglets in binary across experimental group were decreased significantly (P<0.05). In conclusion, Clostridium butyricum can improve intestinal development of piglets, activate TLR2/4 signaling pathway in intestinal mucosa of two genetic background piglets, promote the expression of key negative feedback factors in the pathway, increase the expression of tight junction proteins, decrease the intestinal mucosal permeability, improve intestinal barrier function, and there are certain differences in the regulation of intestinal barrier function of the two genetic background weaned piglets.

Key words： Clostridium butyricum; binary hybridization; ternary hybridization; weaned piglets; Toll-like receptor 2; Toll-like receptor 4

参考文献

[1] 王腾浩.新型丁酸梭菌筛选及其对断奶仔猪生长性能和肠道功能影响的研究[D].博士学位论文.杭州:浙江大学,2015.
[2] LI H H,LI Y P,ZHU Q,et al.Dietary supplementation with Clostridium butyricum helps to improve the intestinal barrier function of weaned piglets challenged with enterotoxigenic Escherichia coli K88[J].Journal of Applied Microbiology,2018,125(4):964-975.

[3] 李玉鹏,李海花,王柳懿,等.丁酸梭菌对断奶仔猪生长性能、肠道屏障功能和血清细胞因子含量的影响[J].动物营养学报,2017,29(8):2961-2968.
[4] 田松军,魏小红,顾丽菊,等.宗地花猪二元和三元杂交猪生产性能的研究[J].黑龙江畜牧兽医,2017(2):63-65.
[5] FUENTES V,VENTANAS S,VENTANAS J,et al.The genetic background affects composition,oxidative stability and quality traits of Iberian dry-cured hams:purebred Iberian versus reciprocal Iberian×Duroc crossbred pigs[J].Meat Science,2014,96:737-743.
[6] XU X F,XU P P,MA C,et al.Gut microbiota,host health,and polysaccharides[J].Biotechnology Advances,2013,31(2):318-337.

[7] 陈秀扬.不同杂交组合的肉猪生产性能及经济效益对比试验[J].广西畜牧兽医,2014(3):133-135.
[8] 孙静泉.杜长大三元杂交猪和长大二元杂交猪生产性能及胴体性状的测定[J].当代畜牧,2010(7):42-43.
[9] LI H H,HANG L,CHEN L B,et al.Lactobacillus acidophilus alleviates the inflammatory response to enterotoxigenic Escherichia coli K88 via inhibition of the NF-κB and P38 mitogen-activated protein kinase signaling pathways in piglets[J].BMC Microbiology,2016,16:273.
[10] 郭军蕊,董晓芳,佟建明.枯草芽孢杆菌联合苜蓿多糖对蛋鸡生产性能、蛋品质、血液指标及粪和肠道微生物区系的影响[J].动物营养学报,2017,29(5):1687-1703.
[11] TAMAGAWA-MINEOKA R,KATOH N.Platelets[M]//KABASHIMA K.Immunology of the skin.Tokyo:Springer,2016:213-226.
[12] ZONG X,WANG T H,LU Z Q,et al.Effects of Clostridium butyricum or in combination with Bacillus licheniformis on the growth performance,blood indexes,and intestinal barrier function of weanling piglets[J].Livestock Science,2019,220:137-142.
[13] YANG K M,JIANG Z Y,ZHENG C T,et al.Effect of Lactobacillus plantarum on diarrhea and intestinal barrier function of young piglets challenged with enterotoxigenic Escherichia coli K88[J].Journal of Animal Science,2014,92(4):1496-1503.

[14] ZHAO L,LUO L,JIA W K,et al.Serum diamine oxidase as a hemorrhagic shock biomarker in a rabbit model[J].PLoS One,2014,9(8):e102285.
[15] ROBINSON K,DENG Z,HOU Y Q,et al.Regulation of the intestinal barrier function by host defense peptides[J].Frontiers in Veterinary Science,2015,2:57.
[16] YANG C M,CAO G T,FERKET P R,et al.Effects of probiotic,Clostridium butyricum,on growth performance,immune function,and cecal microflora in broiler chickens[J].Poultry Science,2012,91(9):2121-2129.

[17] ULLUWISHEWA D,ANDERSON R C,MCNABB W C,et al.Regulation of tight junction permeability by intestinal bacteria and dietary components[J].The Journal of Nutrition,2011,141(5):769-776.

[18] MAO X B,GU C S,HU H Y,et al.Dietary Lactobacillus rhamnosus GG supplementation improves the mucosal barrier function in the intestine of weaned piglets challenged by porcine rotavirus[J].PLoS One,2016,11(1):e0146312.
[19] YANG F J,WANG A N,ZENG X F,et al.Lactobacillus reuteri I5007 modulates tight junction protein expression in IPEC-J2 cells with LPS stimulation and in newborn piglets under normal conditions[J].BMC Microbiology,2015,15:32.
[20] WU Y P,ZHU C,CHEN Z,et al.Protective effects of Lactobacillus plantarum on epithelial barrier disruption caused by enterotoxigenic Escherichia coli in intestinal porcine epithelial cells[J].Veterinary Immunology and Immunopathology,2016,172:55-63.
[21] LI H H,ZHANG L,CHEN L B,et al.Lactobacillus acidophilus alleviates the inflammatory response to enterotoxigenic Escherichia coli K88 via inhibition of the NF-κB and p38 mitogen-activated protein kinase signaling pathways in piglets[J].BMC Microbiology,2016,16:273.
[22] ENSIGN S P F,ROOS A,MATHEWS I T,et al.SGEF is regulated via TWEAK/Fn14/NF-κB signaling and promotes survival by modulation of the DNA repair response to temozolomide[J].Molecular Cancer Research,2016,14(3):302-312.

[23] HUMBERT-CLAUDE M,DUC D,DWIR D,et al.Tollip,an early regulator of the acute inflammatory response in the substantia nigra[J].Journal of Neuroinflammation,2016,13:303.
[24] POVEDA J,SANZ A B,CARRASCO S,et al.Bcl3:a regulator of NF-κB inducible by TWEAK in acute kidney injury with anti-inflammatory and antiapoptotic properties in tubular cells[J].Experimental & Molecular Medicine,2017,49(7):e352.
[25] ROSTAMI K,NEJAD M R,ASADZADEH H,et al.PTU-199 proinflammatory cytokine (IL-8) in microscopic enteritis[J].Gut,2013,62(Suppl.1):A130.

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