动物营养学报 >

2022 , Vol. 34 >Issue 4: 2626 - 2635

DOI: https://doi.org/10.3969/j.issn.1006-267x.2022.04.056

研究论文 RESEARCH PAPER

乳酸锌和硫酸锌对猪肠上皮细胞锌转运载体和屏障功能影响的比较研究

  • 侯若鑫 ,
  • 龙静 ,
  • 彭灿 ,
  • 何流琴 ,
  • 李铁军 ,
  • 汤文杰 ,
  • 邝声耀 ,
  • 印遇龙
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  • 1. 中国科学院亚热带农业生态研究所, 动物营养生理与代谢过程湖南省重点实验室, 长沙 410125;
    2. 中国科学院 大学现代农业科学学院, 北京 100049;
    3. 湖南师范大学生命科学学院, 动物肠道功能调控湖南省重点实验室, 长沙 410081;
    4. 湖南农业大学动物科学技术学院, 长沙 410128;
    5. 四川省畜牧科学研究院, 动物遗传育种 四川省重点实验室, 畜禽生物制品四川省重点实验室, 四川省畜科饲料有限公司, 成都 610066
侯若鑫(1997-),男,湖北襄阳人,硕士研究生,动物营养与饲料科学专业。E-mail:919387379@qq.com

收稿日期: 2021-08-25

  网络出版日期: 2022-04-14

基金资助

广西桂林技术创新引导计划（2020010901）；广西重点研发计划（桂科AB192559012）；国家自然科学基金面上项目（31872371）；四川省应用基础研究（2018JY0641）；湖南省科技领军人才项目（2019RS3022）；广东省基础与应用基础研究基金项目（2019A1515110852）

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Comparative Study on Effects of Zinc Lactate and Zinc Sulfate on Zinc Transporter and Barrier Function in Pig Intestinal Epithelial Cells

  • HOU Ruoxin ,
  • LONG Jing ,
  • PENG Can ,
  • HE Liuqin ,
  • LI Tiejun ,
  • TANG Wenjie ,
  • KUANG Shengyao ,
  • YIN Yulong
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  • 1. Hunan Provincial Key Laboratory of Animal Nutritional Physiology and Metabolic Process, Institute of Subtropical Agriculture, Chinese Academy of Sciences, Changsha 410125, China;
    2. College of Advanced Agricultural Sciences, University of Chinese Academy of Sciences, Beijing 100049, China;
    3. Hunan Province Key Laboratory of Animal Intestinal Function and Regulation, College of Life Sciences, Hunan Normal University, Changsha 410081, China;
    4. School of Animal Science and Technology, Hunan Agricultural University, Changsha 410128, China;
    5. Sichuan Animal Feed Co., Ltd., Sichuan Key Laboratory of Livestock and Poultry Biological Products, Sichuan Key Laboratory of Animal Genetics and Breeding, Sichuan Animal Husbandry Research Institute, Chengdu 610066, China

Received date: 2021-08-25

  Online published: 2022-04-14

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摘要

本研究旨在比较乳酸锌和硫酸锌对仔猪肠上皮细胞IPEC-J2锌转运功能、物理屏障和免疫屏障的影响。将IPEC-J2细胞分别用0、1.0、5.0、7.5、10.0和20.0 mg/mL的乳酸锌或硫酸锌（以锌计）培养36 h，筛选出细胞增殖的最适浓度。根据筛选结果将IPEC-J2细胞随机分为对照组、乳酸锌组和硫酸锌组，每组3个重复。结果表明:1）与对照组相比，7.5 mg/mL乳酸锌和硫酸锌均可显著提高细胞活力（P<0.05），且同等浓度条件下乳酸锌的促进效果更显著。2）与对照组相比，乳酸锌组和硫酸锌组细胞内白细胞介素-1β（IL-1β）和白细胞介素-6（IL-6）含量均显著降低（P<0.05），且硫酸锌组细胞内肿瘤坏死因子-α（TNF-α）含量显著降低（P<0.05），乳酸锌组细胞内TNF-α mRNA相对表达量显著降低（P<0.05）。3）与对照组相比，乳酸锌组细胞内封闭蛋白-1（claudin-1）的蛋白相对表达量显著提高（P<0.05），细胞内E-钙黏蛋白（E-cadherin）的蛋白相对表达量无显著差异（P>0.05）。4）与对照组相比，硫酸锌组细胞内富含半胱氨酸肠蛋白1（CRIP1）和富含半胱氨酸肠蛋白2（CRIP2）含量显著提高（P<0.05），乳酸锌组细胞内CRIP1和CRIP2 mRNA相对表达量显著提高（P<0.05），乳酸锌和硫酸锌组细胞内锌转运蛋白-1（ZNT-1）mRNA相对表达量显著提高（P<0.05）。由此可见，乳酸锌和硫酸锌可抑制IPEC-J2细胞炎性细胞因子的分泌和表达，促进锌的转运，增强肠道紧密连接，从而提高肠道免疫功能和黏膜结构的完整性，改善肠道屏障功能，且乳酸锌效果优于硫酸锌。

关键词： 乳酸锌; IPEC-J2细胞; 屏障功能; 免疫; 锌转运载体

本文引用格式

侯若鑫 , 龙静 , 彭灿 , 何流琴 , 李铁军 , 汤文杰 , 邝声耀 , 印遇龙 . 乳酸锌和硫酸锌对猪肠上皮细胞锌转运载体和屏障功能影响的比较研究[J]. 动物营养学报, 2022 , 34(4) : 2626 -2635 . DOI: 10.3969/j.issn.1006-267x.2022.04.056

Abstract

The aim of this study was to compare the effects of zinc lactate and zinc sulfate on zinc transport function, physical barrier and immune barrier in intestinal epithelial cells IPEC-J2 of piglets. IPEC-J2 cells were cultured with 0, 1.0, 5.0, 7.5, 10.0 and 20.0 mg/mL zinc lactate or zinc sulfate (calculated by zinc) for 36 h, respectively, to screen out the optimal concentration of cell proliferation. Then the IPEC-J2 cells were randomly divided into control group, zinc lactate group and zinc sulfate group according to screening results, with 3 replicates in each group. The results showed as follows:1) compared with the control group, 7.5 mg/mL zinc lactate and zinc sulfate significantly increased the cell viability (P<0.05), and zinc lactate at the same concentration had a more significant promoting effect. 2) Compared with the control group, the contents of interleukin-1β (IL-1β) and interleukin-6 (IL-6) in cells in zinc lactate and zinc sulfate groups were significantly decreased (P<0.05), the tumor necrosis factor-α (TNF-α) content in cells in zinc sulfate group was significantly decreased (P<0.05), and the TNF-α mRNA relative expression level in cells in zinc lactate group was significantly decreased (P<0.05). 3) Compared with the control group, the occludin-1 protein relative expression level in cells in zinc lactate group was significantly increased (P<0.05), while the E-cadherin protein relative expression level in cells was not significantly different (P>0.05). 4) Compared with the control group, the contents of cysteine-rich intestinal protein-1 (CRIP1) and cysteine-rich intestinal protein 2 (CRIP2) in cells in zinc sulfate group were significantly increased (P<0.05), the mRNA relative expression levels of CRIP1 and CRIP2 in cells in zinc lactate group were significantly increased (P<0.05), and the mRNA relative expression level of zinc transporter 1 (ZNT-1) in cells in zinc lactate and zinc sulfate groups was significantly increased (P<0.05). In conclusion, zinc lactate and zinc sulfate can inhibit the secretion and expression of inflammatory cytokines in IPEC-J2 cells, promote the transport of zinc, enhance the intestinal tight junction, thus improve the intestinal immune function and mucosal structure integrity, improve the intestinal barrier function, and the effect of zinc lactate is better than that of zinc sulfate.

Key words： zinc lactate; IPEC-J2 cells; barrier function; immunity; zinc transporter

参考文献

[1] 张锦秀,李书伟,刁慧,等.有机锌对动物肠道健康的调控作用研究进展[J].中国畜牧杂志,2021,57(1):49-53. ZHANG J X,LI S W,DIAO H,et al.Research progress on regulation of organic zinc on animal intestinal health[J].Chinese Journal of Animal Science,2021,57(1):49-53.(in Chinese)
[2] 尹蓝梅.高蛋白饲粮对仔猪肠道免疫及屏障功能的影响研究[D].硕士学位论文.长沙:湖南师范大学,2019. YIN L M.The effects of dietary high protein on intestinal immunity and barrier function in piglets[D].Master’s Thesis.Changsha:Hunan Normal University,2019.(in Chinese)
[3] 叶晓琳,孙梅.肠上皮屏障在炎症性肠病发病机制中的作用[J].中国小儿急救医学,2018,25(9):698-702. YE X L,SUN M.The role of intestinal epithelial barrier in the pathogenesis of inflammatory bowel disease[J].Chinese Pediatric Emergency Medicine,2018,25(9):698-702.(in Chinese)
[4] 郭晓波.海藻多糖对仔猪小肠上皮细胞屏障功能的调控作用[D].硕士学位论文.南昌:江西农业大学,2019. GUO X B.Regulation of seaweed polysaccharide on the barrier function of intestinal porcine epithelial cells[D].Master’s Thesis.Nanchang:Jiangxi Agricultural University,2019.(in Chinese)
[5] IBS K H,RINK L.Zinc-altered immune function[J].The Journal of Nutrition,2003,133(5 Suppl 1):1452S-1456S.
[6] MCDONALD C M,MANJI K P,KISENGE R,et al.Daily zinc but not multivitamin supplementation reduces diarrhea and upper respiratory infections in Tanzanian infants:a randomized,double-blind,placebo-controlled clinical trial[J].The Journal of Nutrition,2015,145(9):2153-2160.  
[7] FINAMORE A,MASSIMI M,CONTI DEVIRGILIIS L,et al.Zinc deficiency induces membrane barrier damage and increases neutrophil transmigration in Caco-2 cells[J].The Journal of Nutrition,2008,138(9):1664-1670.  
[8] 刁慧,张锦秀,晏家友,等.锌对断奶仔猪肠道屏障功能的影响[J].饲料工业,2020,41(19):12-17. DIAO H,ZHANG J X,YAN J Y,et al.Effects of zinc on intestinal barrier function of weaned piglets[J].Feed Industry,2020,41(19):12-17.(in Chinese)
[9] SHI H N,SCOTT M E,STEVENSON M M,et al.Energy restriction and zinc deficiency impair the functions of murine T cells and antigen-presenting cells during gastrointestinal nematode infection[J].The Journal of Nutrition,1998,128(1):20-27.  
[10] BAO S Y,KNOELL D L.Zinc modulates cytokine-induced lung epithelial cell barrier permeability[J].American Journal of Physiology:Lung Cellular and Molecular Physiology,2006,291(6):L1132-L1141.
[11] 俞成浩,郭志强,雷岷,等.乳酸锌对断奶肉兔生长性能、肠道发育和血清生化指标的影响[J].动物营养学报,2021,33(2):1055-1062. YU C H,GUO Z Q,LEI M,et al.Effects of zinc lactate on growth performance,intestinal development and serum biochemical indexes of weaned rabbits[J].Chinese Journal of Animal Nutrition,2021,33(2):1055-1062.(in Chinese)
[12] 王荣蛟,信爱国,张春勇,等.乳酸锌对仔猪小肠形态学、小肠黏膜金属硫蛋白1和组织急性期蛋白mRNA表达的影响[J].动物营养学报,2014,26(4):1068-1076. WANG R J,XIN A G,ZHANG C Y,et al.Effects of zinc lactate on intestinal morphology,mRNA expressions of metallothioneins 1 in intestinal mucosa and acute phase proteins in tissues of weaner piglets[J].Chinese Journal of Animal Nutrition,2014,26(4):1068-1076.(in Chinese)
[13] 王彬,刘路杰,祝佳,等.纳米氧化锌对断奶仔猪生长性能、血清免疫和生化指标的影响[J].动物营养学报,2016,28(11):3626-3633. WANG B,LIU L,ZHU J,et al.Effects of zinc oxide nanoparticles on growth performance,serum immune and biochemical indices of weaned piglets[J].Chinese Journal of Animal Nutrition,2016,28(11):3626-3633.(in Chinese)
[14] 韩国全,余冰,陈代文,等.乳酸锌对猪空肠上皮细胞IPEC-J2增殖及相关调控基因mRNA表达的影响[J].畜牧兽医学报,2012,43(5):740-747. HAN G Q,YU B,CHEN D W,et al.Effect of zinc lactate on cell proliferation and related-regulatory genes mRNA expression in porcine jejunal epithelial cells IPEC-J2[J].Acta Veterinaria et Zootechnica Sinica,2012,43(5):740-747.(in Chinese)
[15] 洪树涛.在体外镉与DNA相互作用:镉结合的协同性和钙、镁、锌竞争的拮抗性[J].国外医学(卫生学分册),1986(1):44-45. HONG S T.Cadmium-DNA interactions in vitro:synergism of cadmium-binding and antagonism to calcium,magnesium,and zinc competition[J].Foreign Medical Sciences (Section Hygiene),1986(1):44-45.(in Chinese)
[16] INAI T,KOBAYASHI J,SHIBATA Y.Claudin-1 contributes to the epithelial barrier function in MDCK cells[J].European Journal of Cell Biology,1999,78(12):849-855.  
[17] MA T M,XU N,MA X D,et al.Moxibustion regulates inflammatory mediators and colonic mucosal barrier in ulcerative colitis rats[J].World Journal of Gastroenterology,2016,22(8):2566-2575.  
[18] 万妍,邵玉新,张炳坤.不同阶段日粮锌营养对肉鸡生长性能及肠黏膜屏障功能的影响[J].中国家禽,2020,42(9):54-61. WAN Y,SHAO Y X,ZHANG B K.Effects of dietary zinc nutrition on growth performance and intestinal mucosal barrier function of chickens at different phase[J].China Poultry,2020,42(9):54-61.(in Chinese)
[19] ZHANG B K,GUO Y M.Supplemental zinc reduced intestinal permeability by enhancing occludin and zonula occludens protein-1(ZO-1) expression in weaning piglets[J].British Journal of Nutrition,2009,102(5):687-693.  
[20] DAVIN R,张永刚.猪小肠上皮细胞作用的新发现[J].中国畜牧杂志,2015,51(8):57-60. DAVIN R,ZHANG Y G.Importance of intestinal cells forgut health-IPEC studies[J].Chinese Journal of Animal Science,2015,51(8):57-60.(in Chinese)
[21] 马茂涛.牡蛎多糖螯合锌的制备螯合及其对脂多糖刺激断奶仔猪小肠健康的影响[D].硕士学位论文.福州:福建农林大学,2017. MA M T.Preparation of zinc-oyster polysaccharide and the effect to lipid stimulation of small intestine health of weaned piglets[D].Master’s Thesis.Fuzhou:Fujian Agriculture and Forestry University,2017.(in Chinese)
[22] ZHAO S W,LI B,CHEN G T,et al.Preparation,characterization,and anti-inflammatory effect of the chelate of Flammulina velutipes polysaccharide with Zn[J].Food and Agricultural Immunology,2017,28(1):162-177.  
[23] THINGHOLM T E.RÖNNSTRAND L,ROSENBERG P A.Why and how to investigate the role of protein phosphorylation in ZIP and ZnT zinc transporter activity and regulation[J].Cellular and Molecular Life Sciences,2020,77(16):3085-3102.  
[24] SHUSTERMAN E,BEHARIER O,LEVY S,et al.Zinc transport and the inhibition of the L-type calcium channel are two separable functions of ZnT-1[J].Metallomics,2017,9(3):228-238.  
[25] ESCOBAR O,SANDOVAL M,VARGAS A,et al.Role of metallothionein and cysteine-rich intestinal protein in the regulation of zinc absorption by diabetic rats[J].Pediatric Research,1995,37(3):321-327.  
[26] 李明月.CRIP1与相关疾病关联性的研究进展[J].当代医药论丛,2019,17(23):83-84. LI M Y.Research progress of CRIP1 and related diseases[J].Contemporary Medicine Forum,2019,17(23):83-84.(in Chinese)
[27] DAVIS B A,BLANCHARD R K,LANNINGHAM-FOSTER L,et al.Structural characterization of the rat cysteine-rich intestinal protein gene and overexpression of this LIM-only protein in transgenic mice[J].DNA and Cell Biology,1998,17(12):1057-1064.  
[28] CAI H M,CHEN J W,LIU J,et al.CRIP1,a novel immune-related protein,activated by Enterococcus faecalis in porcine gastrointestinal epithelial cells[J].Gene,2017,598:84-96.
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