动物营养学报 >

2021 , Vol. 33 >Issue 7: 3745 - 3758

DOI: https://doi.org/10.3969/j.issn.1006-267x.2021.07.017

猪营养与饲料 SWINE NUTRITION AND FEED

早期粪菌移植对仔猪肠道发育、肠道菌群组成和肠道激素分泌的影响

  • 张卓 ,
  • 黄金秀 ,
  • 杨飞云 ,
  • 兰云贤 ,
  • 朱思源 ,
  • 刘静波 ,
  • 刘作华 ,
  • 齐仁立
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  • 1. 西南大学动物科学技术学院, 重庆 402460;
    2. 重庆市畜牧科学院, 重庆 402460;
    3. 农业农村部养猪科学重点实验室, 重庆 402460;
    4. 西南科技大学生命科学与工程学院, 绵阳 621010
张卓(1995-),男,山东冠县人,硕士研究生,从事单胃动物营养研究。E-mail:zzhuo1995@163.com

收稿日期: 2020-12-13

  网络出版日期: 2021-07-06

基金资助

国家重点研发计划项目（2017YFD0500501）；重庆市财政专项资金项目（19513）；重庆荣昌农牧高新技术专项项目（20206）

收起

Effects of Early Fecal Microbiota Transplantation on Intestinal Development, Intestinal Microbiota Composition and Intestinal Hormone Secretion of Piglets

  • ZHANG Zhuo ,
  • HUANG Jinxiu ,
  • YANG Feiyun ,
  • LAN Yunxian ,
  • ZHU Siyuan ,
  • LIU Jingbo ,
  • LIU Zuohua ,
  • QI Renli
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  • 1. College of Animal Science and Technology, Southwest University, Chongqing 402460, China;
    2. Chongqing Academy of Animal Science, Chongqing 402460, China;
    3. Key Laboratory of Pig Industry Sciences, Ministry of Agriculture, Chongqing 402460, China;
    4. School of Life Science and Engineering, Southwest University of Science and Technology, Mianyang 621010, China

Received date: 2020-12-13

  Online published: 2021-07-06

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摘要

本试验旨在研究早期粪菌移植（FMT）对仔猪肠道发育、肠道菌群组成、肠道激素分泌的影响。选取相同出生日期的新生"长×荣"二杂仔猪6窝，分为2组，每组3窝。在出生后3~7 d，FMT组仔猪每天灌喂1 mL健康成年猪的粪便菌悬液，对照组仔猪灌喂等量无菌生理盐水。28日龄时，每组挑选接近平均体重的6头健康仔猪采集静脉血后屠宰，收集相应肠道及内容物样品用于组织形态、激素水平、基因表达和肠道菌群组成的检测。结果表明：1）与对照组仔猪相比，FMT组仔猪平均日增重（ADG）提高了约20%（P<0.01），十二指肠绒毛高度极显著降低（P<0.01），而隐窝深度极显著增加（P<0.01）。2）与对照组仔猪相比，FMT组仔猪血清中胆囊收缩素（CCK）、胰高血糖素样肽-1（GLP-1）和饥饿素（ghrelin）这3种肠道激素的水平不同程度增加，其中GLP-1和ghrelin的水平达到显著水平（P<0.05）；FMT组仔猪回肠中CCK和ghrelin表达水平显著或极显著增加（P<0.05或P<0.01），CCK的受体胆囊收缩素A受体（CCKAR）和胆囊收缩素B受体（CCKBR）的mRNA相对表达量同样显著或极显著增加（P<0.05或P<0.01），而ghrelin的受体生长激素促分泌素受体（GHSR）的mRNA相对表达量无显著变化（P>0.05）；FMT组仔猪结肠中GLP-1的表达水平极显著升高（P<0.01），其受体胰高血糖素样肽-1受体（GLP-1R）的mRNA相对表达量有上升趋势（P>0.05）。3）FMT明显改变了受体仔猪回肠中的菌群结构，增加了结肠菌群的alpha多样性。在门水平上，与对照组仔猪相比，FMT组仔猪回肠中厚壁菌门（Firmicutes）的相对丰度增加，而变形菌门（Proteobacteria）的相对丰度减少；在属水平上，与对照组仔猪相比，FMT组仔猪回肠中乳杆菌属（Lactobacillus）的相对丰度减少，SMB53、Sarcina和棒状杆菌属（Corynebacterium）的相对丰度增加。2组仔猪的结肠菌群组成差异不明显。4）将肠道中相对丰度排在前3位的菌群与肠道激素表达水平进行Spearman相关性分析，发现回肠中韦荣球菌属（Veillonella）和嗜血杆菌属（Haemophilus）的相对丰度与GLP-1表达水平呈显著负相关（P<0.05）；结肠中颤螺菌属（Oscillospira）的相对丰度与GLP-1表达水平呈极显著正相关（P<0.01），密螺旋体属（Treponema）的相对丰度与CCK表达水平呈显著正相关（P<0.05），而瘤胃球菌属（Ruminococcus）和拟杆菌属（Bacteroides）的相对丰度与CCK表达水平呈显著负相关（P<0.05）。这些结果表明早期FMT能够加速受体仔猪的生长和肠道发育，并通过干预肠道菌群组成的变化影响肠道激素的分泌和传递。

关键词： 粪菌移植; 仔猪; 肠道激素; 胆囊收缩素; 胰高血糖素样肽-1; 饥饿素

本文引用格式

张卓 , 黄金秀 , 杨飞云 , 兰云贤 , 朱思源 , 刘静波 , 刘作华 , 齐仁立 . 早期粪菌移植对仔猪肠道发育、肠道菌群组成和肠道激素分泌的影响[J]. 动物营养学报, 2021 , 33(7) : 3745 -3758 . DOI: 10.3969/j.issn.1006-267x.2021.07.017

Abstract

The aim of this present study was to investigate the effects of early fecal microbiota transplantation (FMT) on intestinal development, intestinal microbiota composition and intestinal hormone secretion of piglets. Six litters of newborn "Landrace×Rongchang" newborn piglets with the same birth date were selected and divided into two groups with three litters in each group. The piglets in FMT group were given 1 mL of fecal microbiota suspension of healthy adult pigs every day at 3 to 7 days after birth, while the piglets in control group were given an equal volume of sterile saline. At 28 days of age, six healthy piglets with average body weight were selected and sacrificed. Their blood, intestine tissue and intestinal digesta were collected for the assay of intestinal morphology, hormone levels, gene expression and intestinal microbiota composition. The results showed as follows:1) compared with the piglets in control group, the average daily gain (ADG) of piglets in FMT group was increased by about 20% (P<0.01), the duodenal villi height was extremely significantly decreased (P<0.01), while the crypt depth was extremely significantly increased (P<0.01). 2) Compared with the piglets in control group, the levels of three intestinal hormones, cholecystokinin (CCK), glucagon-like peptide-1 (GLP-1) and ghrelin in serum of piglets in FMT group were increased at different degrees, and the GLP-1 and ghrelin reached significant level (P<0.05); the expression levels of CCK and ghrelin in ileum of piglets in FMT group were significantly or extremely significantly increased (P<0.05 or P<0.01), and the mRNA relative expression levels of CCK receptors-cholecystokinin A receptor (CCKAR) and cholecystokinin B receptor (CCKBR) were also significantly or extremely significantly up-regulated (P<0.05 or P<0.01), while ghrelin receptor-growth hormone secretagogue receptor (GHSR) showed a down-regulated trend (P>0.05); the expression level of GLP-1 in colon of piglets in FMT group was extremely significantly increased (P<0.01), however, the mRNA relative expression level of its receptor-glucagon-like peptide-1 receptor (GLP-1R) showed an upward trend (P>0.05). 3) FMT obviously changed intestinal microbial community in the recipient piglets and especially increased the alpha diversity of colonic microbiota. At phylum level, the relative abundance of Firmicutes in ileum of piglets in FMT group was increased, while the relative abundance of Proteobacteria was decreased compared with the piglets in control group. At genus level, the relative abundance of Lactobacillus in ileum of piglets in FMT group was decreased, while that of SMB53, Sarcina and Corynebacterium were increased compared with the piglets in control group. No obvious difference in colonic microbial composition between the two groups of piglets. 4) The Spearman correlation analysis was conducted between the relative abundances of top intestinal microbiota and intestinal hormone expression levels, and it was found that the relative abundances of Veillonella and Haemophilus in ileum were significantly negatively correlated with the expression level of GLP-1 (P<0.05); the relative abundance of Oscillospira in colon was extremely significantly positively correlated with GLP-1 expression level (P<0.01), and the relative abundance of Treponema in colon was significantly positively correlated with CCK expression level (P<0.05), while the relative abundances of Ruminococcus and Bacteroidetes in colon were significantly negatively correlated with CCK expression level (P<0.05). These results reveal that the early FMT can promote the growth and intestinal development of recipient piglets, and can affect the secretions and transmission of intestinal hormones by intervening intestinal microbiota composition.

Key words： fecal microbiota transplantation; piglet; intestinal hormones; cholecystokinin; glucagon-like peptide-1; ghrelin

参考文献

[1] BEUMER J,PUSCHHOF J,BAUZÁ-MARTINEZ J,et al.High-resolution mRNA and secretome atlas of human enteroendocrine cells[J].Cell,2020,181(6):1291-1306.  
[2] 朱翠,杨俊,蒋宗勇,等.仔猪胃肠道内分泌细胞分布与化学感应作用[J].动物营养学报,2018,30(11):4286-4294. ZHU C,YANG J,JIANG Z,et al.Distribution and chemosensing role of enteroendocrine cells in piglets[J].Chinese Journal of Animal Nutrition,2018,30(11):4286-4294.(in Chinese)
[3] 孙晓宇,郭汝涛,张洪涛,等.胃肠激素与肥胖关系的研究进展[J].现代生物医学进展,2015,15(14):2786-2789. SUN X Y,GUO R T,ZHANG H T,et al.Research on correlations of gastrointestinal hormones and calorigenic nutrients for obesity[J].Progress in Modern Biomedicine,2015,15(14):2786-2789.(in Chinese)
[4] 杜莉,卢顺娇,王英超,等.胰高血糖素样肽-1生物学功能的研究进展[J].生物化工,2017,3(6):97-98,104. DU L,LU S J,WANG Y C,et al.Research progress of glucagon like peptide-1 and its biological functions[J].Biochemical Engineering,2017,3(6):97-98,104.(in Chinese)
[5] 孙睿,杨文艳.胃饥饿素分泌调控机制的研究进展[J].动物营养学报,2021,33(1):132-140. SUN R,YANG W Y.Advances on regulation mechanism of ghrelin secretion[J].Chinese Journal of Animal Nutrition,2021,33(1):132-140.(in Chinese)
[6] SHEN J,OBIN M S,ZHAO L P.The gut microbiota,obesity and insulin resistance[J].Molecular Aspects of Medicine,2013,34(1):39-58.  
[7] 王后福,李鹏飞,矣国,等.微生物营养物质代谢与微生物-肠-脑轴互作研究进展[J].动物营养学报,2020,32(1):28-35. WANG H F,LI P F,YI G,et al.Progress in microbial nutrient metabolism and microbiota-gut-brain axis interaction[J].Chinese Journal of Animal Nutrition,2020,32(1):28-35.(in Chinese)
[8] SAMPSON T R,MAZMANIAN S K.Control of brain development,function,and behavior by the microbiome[J].Cell Host & Microbe,2015,17(5):565-576.  
[9] 张亚南,余凯凡,朱伟云.肠道微生物调控宿主食欲的研究进展[J].微生物学报,2017,57(7):951-960. ZHANG Y N,YU K F,ZHU W Y.Role of gut microbiota in host appetite control[J].Acta Microbiologica Sinica,2017,57(7):951-960.(in Chinese)
[10] URIBE A,ALAM M,JOHANSSON O,et al.Microflora modulates endocrine cells in the gastrointestinal mucosa of the rat[J].Gastroenterology,1994,107(5):1259-1269.  
[11] GENSOLLEN T,IYER S S,KASPER D L,et al.How colonization by microbiota in early life shapes the immune system[J].Science,2016,352(6285):539-544.  
[12] NGUYEN Q N,HIMES J E,MARTINEZ D R,et al.The impact of the gut microbiota on humoral immunity to pathogens and vaccination in early infancy[J].PLoS Pathogens,2016,12(12):e1005997.
[13] KORPELA K,DE VOS W M.Early life colonization of the human gut:microbes matter everywhere[J].Current Opinion in Microbiology,2018,44:70-78.
[14] HO J T,CHAN G C,LI J C.Systemic effects of gut microbiota and its relationship with disease and modulation[J].BMC Immunology,2015,16:21.
[15] GENG S J,CHENG S S,LI Y,et al.Faecal microbiota transplantation reduces susceptibility to epithelial injury and modulates tryptophan metabolism of the microbial community in a piglet model[J].Journal of Crohn's and Colitis,2018,12(11):1359-1374.
[16] BRUNSE A,MARTIN L,RASMUSSEN T S,et al.Effect of fecal microbiota transplantation route of administration on gut colonization and host response in preterm pigs[J].The ISME Journal,2019,13(3):720-733.  
[17] NIEDERWERDER M C,CONSTANCE L A,ROWLAND R R R,et al.Fecal microbiota transplantation is associated with reduced morbidity and mortality in porcine circovirus associated disease[J].Frontiers in Microbiology,2018,9:1631.
[18] CHENG C S,WEI H K,WANG P,et al.Early intervention with faecal microbiota transplantation:an effective means to improve growth performance and the intestinal development of suckling piglets[J].Animal,2019,13(3):533-541.  
[19] DIAO H,YAN H L,XIAO Y,et al.Modulation of intestine development by fecal microbiota transplantation in suckling pigs[J].RSC Advances,2018,8(16):8709-8720.  
[20] HU L S,GENG S J,LI Y,et al.Exogenous fecal microbiota transplantation from local adult pigs to crossbred newborn piglets[J].Frontiers in Microbiology,2018,8:2663.
[21] 刘璐.乳源性复合益生菌乳对db/db小鼠抗糖尿病作用及GLP-1的影响[D].硕士学位论文.乌鲁木齐:新疆医科大学,2018. LIU L.Effects of milk-borne composit probiotic on glucagon-like peptide-1 in db/db diabetic mice[D].Master's Thesis.Urumchi:Xinjiang Medical University,2018.(in Chinese)
[22] DUCA F A,SWARTZ T D,SAKAR Y,et al.Increased oral detection,but decreased intestinal signaling for fats in mice lacking gut microbiota[J].PLoS One,2012,7(6):e39748.
[23] PERRY R J,PENG L,BARRY N A,et al.Acetate mediates a microbiome-brain-β-cell axis to promote metabolic syndrome[J].Nature,2016,534(7606):213-217.  
[24] WYNNE K,STANLEY S,MCGOWAN B,et al.Appetite control[J].Journal of Endocrinology,2005,184(2):291-318.  
[25] AHMED M,AHMED S.Functional,diagnostic and therapeutic aspects of gastrointestinal hormones[J].Gastroenterology Research,2019,12(5):233-244.  
[26] BLANCO A M,BERTUCCI J I,VALENCIANO A I,et al.Ghrelin suppresses cholecystokinin (CCK),peptide YY (PYY) and glucagon-like peptide-1(GLP-1) in the intestine,and attenuates the anorectic effects of CCK,PYY and GLP-1 in goldfish (Carassius auratus)[J].Hormones and Behavior,2017,93:62-71.
[27] TREMAROLI V,BÄCKHED F.Functional interactions between the gut microbiota and host metabolism[J].Nature,2012,489(7415):242-249.  
[28] ZELKAS L,RAGHUPATHI R,LUMSDEN A L,et al.Serotonin-secreting enteroendocrine cells respond via diverse mechanisms to acute and chronic changes in glucose availability[J].Nutrition & Metabolism,2015,12:55.
[29] 张贺,徐荣莹,苏勇,等.单胃动物肠道微生物研究进展[J].动物营养学报,2020,32(10):4674-4685. ZHANG H,XU R Y,SU Y,et al.A review:gut microbiota in monogastric animals[J].Chinese Journal of Animal Nutrition,2020,32(10):4674-4685.(in Chinese)
[30] JIANG T T,SHAO T Y,ANG W X G,et al.Commensal fungi recapitulate the protective benefits of intestinal bacteria[J].Cell Host & Microbe,2017,22(6):809-816.  
[31] KAOUTARI A E,ARMOUGOM F,GORDON J I,et al.The abundance and variety of carbohydrate-active enzymes in the human gut microbiota[J].Nature Reviews Microbiology,2013,11(7):497-504.  
[32] GOPHNA U,KONIKOFF T,NIELSEN H B.Oscillospira and related bacteria-from metagenomic species to metabolic features[J].Environmental Microbiology Reports,2017,19(3):835-841.  
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