研究论文 RESEARCH PAPER

屎肠球菌对仔猪小肠紧密连接蛋白、细胞因子应答以及Toll样受体基因表达的影响

  • 崔百磊 ,
  • 王丽 ,
  • 郭乾鹏 ,
  • 黄怡 ,
  • 胡胜兰
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  • 1. 广西大学动物科学技术学院, 南宁 530004;
    2. 广东省农业科学院动物科学研究所, 畜禽育种国家重点实验室, 农业农村部华南动物营养与饲料重点实验室, 广东省畜禽育种与营养研究重点实验室, 岭南现代农业 科学与技术广东省实验室茂名分中心, 广州 510640
崔百磊(1997—),男,江苏如皋人,硕士研究生,研究方向为猪营养与饲料。E-mail:1308573974@qq.com

收稿日期: 2022-05-11

  网络出版日期: 2022-12-15

基金资助

国家自然科学基金项目(31460607);广东省农业科学院十四五杰出团队项目(202106TD)

Effects of Enterococcus faecium on Expression of Intestinal Tight Junction Proteins, Cytokine Responses and Toll-Like Receptors Genes in Piglets

  • CUI Bailei ,
  • WANG Li ,
  • GUO Qianpeng ,
  • HUANG Yi ,
  • HU Shenglan
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  • 1. College of Animal Science and Technology, Guangxi University, Nanning 530004, China;
    2. Maoming Branch, Guangdong Laboratory for Lingnan Modern Agriculture, Guangdong Key Laboratory of Animal Breeding and Nutrition, Key Laboratory of Animal Nutrition and Feed Science in South China Ministry of Agriculture, State Key Laboratory of Livestock and Poultry Breeding, Institute of Animal Science, Guangdong Academy of Agricultural Sciences, Guangzhou 510640, China

Received date: 2022-05-11

  Online published: 2022-12-15

摘要

本研究旨在比较屎肠球菌活菌和灭活菌对仔猪小肠紧密连接蛋白、细胞因子应答以及Toll样受体(TLR)基因表达的影响。试验选用健康的杜×长×大新生仔猪9窝,窝产仔数10~11头,随机分为对照组(CON组)、屎肠球菌活菌组(LB组)和屎肠球菌灭活菌组(HB组),每组3窝仔猪。从出生开始至6日龄,CON组、LB组和HB组仔猪分别经口灌服无菌脱脂乳、含屎肠球菌活菌(1×108 CFU/mL)的脱脂乳和含屎肠球菌灭活菌(1×108 CFU/mL)的脱脂乳,口服剂量为10 mL/(头·d)。仔猪于7日龄开始饲喂教槽料,21日龄断奶,并分别于这2个日龄从每窝中选出1头接近平均体重的仔猪屠宰取样,检测小肠黏膜紧密连接蛋白和细胞因子应答基因表达及分泌量,以及Toll样受体基因的表达。结果表明:1)与CON组相比,LB组7日龄仔猪空肠黏膜闭锁小带蛋白-1(ZO-1)基因相对表达量有提高趋势(P=0.08),而回肠黏膜ZO-1基因相对表达量则显著提高(P<0.05);LB组和HB组21日龄仔猪空肠和回肠黏膜ZO-1基因相对表达量均无显著变化(P>0.05)。2)与CON组相比,仔猪7日龄时,LB组空肠黏膜白细胞介素-8(IL-8)基因相对表达量有降低趋势(P=0.052),白细胞介素-10(IL-10)基因相对表达量呈提高趋势(P=0.063),但肿瘤坏死因子-α(TNF-α)和转化生长因子-β(TGF-β)基因相对表达量均无显著变化(P>0.05);HB组空肠黏膜IL-8基因相对表达量显著降低(P<0.05),但其他细胞因子基因相对表达量均无显著变化(P>0.05);LB组和HB组回肠黏膜IL-8、IL-10、TNF-αTGF-β基因相对表达量均无显著变化(P>0.05)。与CON组相比,仔猪21日龄时,LB组空肠黏膜IL-8基因相对表达量显著降低(P<0.05),回肠黏膜IL-8基因相对表达量也显著降低(P<0.05),且IL-10和TGF-β基因相对表达量显著提高(P<0.05);HB组空肠黏膜IL-8基因相对表达量显著降低(P<0.05),TGF-β基因相对表达量显著提高(P<0.05),但回肠黏膜的4种细胞因子基因相对表达量无显著差异(P>0.05)。3)与CON组相比,LB组7日龄仔猪空肠黏膜TNF-α分泌量有降低趋势(P=0.087),而回肠黏膜IL-10分泌量则显著提高(P<0.05),且21日龄仔猪空肠黏膜IL-8分泌量显著减少(P<0.05),对TGF-β的分泌量无显著影响(P>0.05);HB组7日龄仔猪空肠和回肠黏膜细胞因子分泌量均无显著变化(P>0.05)。4)与CON组相比,LB组和HB组7日龄仔猪空肠和回肠黏膜TLR-2、TLR-4、TLR-9基因相对表达量均无显著变化(P>0.05);LB组和HB组21日龄仔猪空肠黏膜TLR-2、TLR-4、TLR-9基因相对表达量也无显著变化(P>0.05),但HB组回肠黏膜TLR-4基因相对表达量显著提高(P<0.05)。综上所述,屎肠球菌活菌比灭活菌更有利于增强小肠黏膜紧密连接蛋白基因表达,调节细胞因子基因表达和分泌,并稳定小肠黏膜TLR基因表达,从而改善仔猪肠道健康。

本文引用格式

崔百磊 , 王丽 , 郭乾鹏 , 黄怡 , 胡胜兰 . 屎肠球菌对仔猪小肠紧密连接蛋白、细胞因子应答以及Toll样受体基因表达的影响[J]. 动物营养学报, 2022 , 34(12) : 7616 -7627 . DOI: 10.3969/j.issn.1006-267x.2022.12.015

Abstract

The objective of this study was to compare the effects of live and heat-inactivated bacteria of Enterococcus faecium on expression of small intestinal tight junction proteins, cytokine responses, and Toll-like receptor (TLR) genes of piglets. Nice litters (10 to 11 piglets per litter) of healthy neonatal piglets (Duroc×Landrace×Yorkshire) were selected and randomly divided into three groups:control group (CON group), E. faecium group (LB group) and heat-inactivated E. faecium group (HB group), with 3 litters in each group. From birth to 6 days of age, piglets from 3 groups were orally fed sterile skimmed milk, skimmed milk containing E. faecium, and skimmed milk containing heat-inactivated E. faecium (concentration 1×108 CFU/mL), 10 mL/(head·d), respectively. Creep feed was added for piglets from 7 days of age to 21 days of age (weaned day). On 7 and 21 days of age, one piglet from each litter was selected for slaughter sampling to detect small intestine mucosa tight junction protein gene expression, cytokine gene expression and secretion, and gene expression of TLR. The results showed as follows:1) compared with the CON group, the relative expression level of zonula occludens-1 (ZO-1) gene in jejunal mucosa of 7-day-old piglets in LB group had a trend to improve (P=0.08), while the ZO-1 gene relative expression level in ileal mucosa was significantly increased (P<0.05); There were no significant changes in relative expression level of ZO-1 gene in the jejunal and ileum of 21-day-old piglets in the LB and HB groups (P>0.05). 2) At 7 days of age, the jejunal mucosal cytokine interleukin (IL)-8 gene relative expression level had a trend to decrease (P=0.052), and the IL-10 gene relative expression level had a trend to increase (P=0.063), however, neither tumor necrosis factor-alpha (TNF-α) nor transforming growth factor-beta (TGF-β) changed significantly (P>0.05). The relative expression level of IL-8 gene in the jejunal mucosa was significantly decreased in the HB group (P<0.05), however, there was no significant change in the relative expression of any other cytokines (P>0.05). There were no significant changes in IL-8、IL-10、TNF-α and TGF-β gene relative expression levels in the ileal mucosa in both LB and HB groups (P>0.05). At 21 days of age, the jejunal and ileal mucosa IL-8 gene relative expression level was significantly decreased (P<0.05), and the ileal mucosa cytokine IL-10 and TGF-β gene relative expression levels were significantly increased (P<0.05). The relative expression level of IL-8 gene in jejunal mucosa in HB group decreased significantly (P<0.05), the relative expression level of TGF-β gene was significantly increased (P<0.05), and the relative expression level of 4 cytokines gene in the ileal mucosa was not significantly changed (P>0.05). 3) The secretion of TNF-α in the jejunum mucosa of 7-day-old piglets in LB group had a trend to decrease (P=0.087), while the secretion of IL-10 in the ileal mucosa was significantly increased (P<0.05), and the content of IL-8 in the jejunal mucosa of 21-day-old piglets was significantly reduced (P<0.05), which had no significant effect on the content of TGF-β (P>0.05). There were no significant changes in mucosal cytokine secretion in the jejunum and ileum of 7-day-old piglets in the HB group (P>0.05). 4) The relative expression levels of TLR-2, TLR-4 and TLR-9 genes were not significantly changed in jejunal of 7-day-old piglets in LB and HB group (P>0.05), and those of TLR-2, TLR-4 and TLR-9 genes were not significantly changed in jejunum of 21-day-old piglets in LB and HB group (P>0.05), however, the relative expression level of TLR-4 gene was significantly increased in HB group (P<0.05). All the above-mentioned results indicate that compared with heat-inactivated E. faecium, live bacteria are more efficient to regulate the intestinal health of piglets by enhancing the expression of tight junction, mucosal cytokines, and stable TLR expression.

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