过量赖氨酸对断奶仔猪及其肠道上皮细胞的影响

doi:10.11843/j.issn.0366-6964.2021.07.014

摘要/Abstract

摘要： 旨在从体内与体外探究过量赖氨酸对断奶仔猪的影响。本试验选用144头杜×长×大三元杂交断奶仔猪，随机分为4组，每组6个重复，每个重复6头仔猪（3头阉公猪和3头母猪），各组在日粮中分别添加1.3%、2.6%、3.9%和5.2%的回肠标准可消化赖氨酸（standardized ileal digestibility lysine，SID Lys），观察其对于仔猪生长性能、器官指数以及生理生化等指标的影响。以IPEC-J2为体外模型，根据培养基与营养需要中氨基酸的种类与比例，添加赖氨酸及其他氨基酸，测定IPEC-J2细胞的增殖情况，并试图通过平衡氨基酸手段降低因过量添加赖氨酸造成的不利影响。结果显示，随着日粮SID Lys添加量的增加，断奶仔猪的生长性能显著下降（P<0.05），血浆平均红细胞血红蛋白含量（mean corpuscular hemoglobin，MCH）、部分血清游离氨基酸含量和肠道形态受到影响。当在培养基中添加2.0 mmol·L^-1赖氨酸时，IPEC-J2细胞活力显著下降（P<0.05），培养基中原赖氨酸浓度（0.5 mmol·L^-1）是IPEC-J2生长的最适浓度。当按照培养基中氨基酸平衡比例补充其他必需氨基酸时，细胞活力有一定程度的改善；当按照猪碱性氨基酸的最适比例补充相应浓度的精氨酸与组氨酸时，细胞活力随处理时间的变化而变化。综上所述，单独添加过多的赖氨酸对断奶仔猪的生长会产生负面作用，在生产实际应用中应充分考虑氨基酸之间的平衡。

关键词: 赖氨酸, 断奶仔猪, IPEC-J2

Abstract: The aim of this study was to investigate the effects of excess lysine on weaned piglets in vivo and in vitro. A total of 144 weaned piglets (Duroc×Landrace×Yorkshire) were randomly divided into 4 groups, 6 replicates for each group, 6 piglets for each replicate (3 barrows and 3 sows). The pigs in different groups were fed with excessive standardized ileal digestibility lysine (SID Lys) of 1.3%, 2.6%, 3.9% and 5.2%, respectively in the diet to observe its effects on growth performance, visceral indexes and physiological and biochemical indicators. Using IPEC-J2 as an in vitro model, the proliferation of IPEC-J2 cells was measured by adding lysine and other amino acids according to the type and ratio of amino acids in the culture medium and nutritional requirements, and attempting to reduce the adverse effects of excessive addition of lysine by balancing other amino acids. The results showed that, with the increase of the amount of SID Lys in the diets, the growth performance of weaned piglets significantly decreased (P<0.05), the mean corpuscular hemoglobin (MCH), some serum free amino acids and intestinal morphology were affected. When lysine was 2.0 mmol·L^-1in culture medium, the viability of IPEC-J2 cells significantly decreased(P<0.05), with the original lysine concentration in the medium (0.5 mmol·L^-1) being the optimum concentration for IPEC-J2 growth. Cell viability was improved to some extent when other essential amino acids were supplemented according to the balanced ratio of amino acids in the medium; Cell viability varied with treatment time when the corresponding concentrations of arginine and histidine were supplemented according to the optimum ratio of porcine basic amino acids. The study demonstrated that the addition of excessive lysine has negative effect on the growth of weaned piglets. The balance of amino acids should be fully considered in practical applications.

Key words: lysine, weaned piglets, IPEC-J2

中图分类号:

S816
S828.5

贾红敏, 马永航, 贺平丽, 谯仕彦. 过量赖氨酸对断奶仔猪及其肠道上皮细胞的影响[J]. 畜牧兽医学报, 2021, 52(7): 1912-1926.

JIA Hongmin, MA Yonghang, HE Pingli, QIAO Shiyan. Effects of Excessive Lysine on Weaned Piglets and Their Intestinal Epithelial Cells[J]. Acta Veterinaria et Zootechnica Sinica, 2021, 52(7): 1912-1926.

参考文献 43

[1]	ROY N,LAPIERRE H,BERNIER J F.Whole-body protein metabolism and plasma profiles of amino acids and hormones in growing barrows fed diets adequate or deficient in Lysine[J].Can J Anim Sci,2000,80(4):585-595.
[2]	REN J B,ZHAO G Y,LI Y X,et al.Influence of dietary Lysine level on whole-body protein turnover,plasma IGF-I,GH and insulin concentration in growing pigs[J].Livest Sci,2007,110(1-2):126-132.
[3]	罗钧秋,陈代文.赖氨酸对蛋白质代谢的影响及其可能调控机制[J].饲料工业,2006,27(16):40-43.LUO J Q,CHEN D W.Effect of lysine on protein metabolize and its possibility mechanism of regulation and control[J].Feed Industry,2006,27(16):40-43.(in Chinese)
[4]	BAKER D H.Advances in protein-amino acid nutrition of poultry[J].Amino Acids,2009,37(1):29-41.
[5]	FAURE M,MOËNNOZ D,MONTIGON F,et al.Dietary threonine restriction specifically reduces intestinal mucin synthesis in rats[J].J Nutr,2005,135(3):486-491.
[6]	RHOADS J M,WU G Y.Glutamine,arginine,and leucine signaling in the intestine[J].Amino Acids,2009,37(1):111-122.
[7]	TAN B,YIN Y L,KONG X F,et al.L-arginine stimulates proliferation and prevents endotoxin-induced death of intestinal cells[J].Amino Acids,2010,38(4):1227-1235.
[8]	WU G Y.Functional amino acids in growth,reproduction,and health[J].Adv Nutr,2010,1(1):31-37.
[9]	TORRAS-LLORT M,SORIANO-GARCÍA J F,FERRER R,et al.Effect of a Lysine-enriched diet on L-Lysine transport by the brush-border membrane of the chicken jejunum[J].Am J Physiol,1998,274(1):R69-R75.
[10]	宋娇.葡萄糖和氨基酸通过mTORC1信号通路影响小鼠皮肤成纤维细胞生长[D].重庆:重庆医科大学,2013.SONG J.Glucose and leucine through the mTORC1 signal pathway affect the fibroblasts growth[D]. Chongqing:Chongqing Medical University,2013.(in Chinese)
[11]	夏添.不同锌源调控断奶仔猪腹泻机理的研究[D].北京:中国农业大学,2017.XIA T.Study on different zinc sources in modulating mechanisms of reducing incidence of diarrhea of weaned piglets[D].Beijing:China Agricultural University,2017.(in Chinese)
[12]	中华人民共和国国家质量监督检验检疫总局,中国国家标准化管理委员会.GB/T 6435-2014饲料中水分的测定[S].北京:中国标准出版社,2015.General Administration of Quality Supervision,Inspection and Quarantine of the People's Republic of China, Standardization Administration.GB/T 6435-2014 Determination of moisture in feedstuffs[S].Beijing:Standards Press of China,2015.(in Chinese)
[13]	国家市场监督管理总局,国家标准化管理委员会.GB/T 6432-2018饲料中粗蛋白的测定凯氏定氮法[S].北京:中国标准出版社,2018.State Administration for Market Regulation,Standardization Administration.GB/T 6432-2018.Determination of crude protein in feeds-Kjeldahl method[S].Beijing:Standards Press of China,2018.(in Chinese)
[14]	国家市场监督管理总局,国家标准化管理委员会.GB/T 6437-2018饲料中总磷的测定分光光度法[S].北京:中国标准出版社,2018.State Administration for Market Regulation,Standardization Administration.GB/T 6437-2018 Determination of phosphorus in feeds-Spectrophotometry[S].Beijing:Standards Press of China,2018.(in Chinese)
[15]	国家市场监督管理总局,中国国家标准化管理委员会.GB/T 6436-2018饲料中钙的测定[S].北京:中国标准出版社,2018.State Administration for Market Regulation,Standardization Administration.GB/T 6436-2018 Determination of calcium in feeds[S].Beijing:Standards Press of China,2018.(in Chinese)
[16]	中华人民共和国国家市场监督管理总局,中国国家标准化管理委员会.GB/T 18246-2019饲料中氨基酸的测定[S].北京:中国标准出版社,2019.State Administration for Market Regulation,Standardization Administration.GB/T 18246-2019 Determination of amino acids in feeds[S].Beijing:Standards Press of China,2019.(in Chinese)
[17]	国家市场监督管理总局,国家标准化管理委员会.GB/T 15399-2018饲料中含硫氨基酸的测定离子交换色谱法[S].北京:中国标准出版社,2018.State Administration for Market Regulation,Standardization Administration.GB/T15399-2018 Determination of sulfur amino acids in feeds-Ion exchange chromatography[S].Beijing:Standards Press of China,2018.(in Chinese)
[18]	方晨,郭飞,胡瑞举,等.杂交组合断奶仔猪腹泻与FUT1基因遗传变异的关联分析[J].畜牧兽医学报,2021,52(3):610-619.FANG C,GUO F,HU R J,et al.Analysis of correlation between diarrhea and genetic variation of FUT1 gene in weaned piglets of hybrid combination[J].Acta Veterinaria et Zootechnica Sinica,2021,52(3):610-619.(in Chinese)
[19]	张振宇,梁春年,姚喜喜,等.日粮不同营养水平对牦牛生产性能、屠宰指标和血清生化指标的影响[J].畜牧兽医学报,2021,52(1):135-143.ZHANG Z Y,LIANG C N,YAO X X,et al.Effects of different nutrition levels of diets on production performance, slaughter indexes and serum biochemical indexes of yak[J].Acta Veterinaria et Zootechnica Sinica, 2021, 52(1):135-143.(in Chinese)
[20]	HAHN J D,BAKER D H.Optimum ratio to lysine of threonine,tryptophan,and sulfur amino acids for finishing swine[J].J Anim Sci,1995,73(2):482-489.
[21]	李德发,顾宏伟,谯仕彦,等.日粮赖氨酸水平对生长肥育猪氮代谢的影响[J].中国饲料,1992(5):15-17.LI D F,GU H W,QIAO S Y,et al.Effects of dietary lysine level on nitrogen metabolism of growing finishing pigs[J].China Feed,1992(5):15-17.(in Chinese)
[22]	谯仕彦,李德发,张国龙,等.增加日粮赖氨酸对断奶仔猪生长、氮存留和血浆游离氨基酸含量的影响[J].中国畜牧杂志,1996,32(3):3-6.QIAO S Y,LI D F,ZHANG G L,et al.Effect of increased dietary lysine level on performance N retention and plasma amino acid profiles in weanling pigs[J].Chinese Journal of Animal Science,1996,32(3):3-6.(in Chinese)
[23]	谢建兵,LEE H C,段红伟,等.饲粮不同赖氨酸水平对早期断奶仔猪生产性能的影响[J].饲料工业,2010,31(20):5-7.XIE J B,LEE H C,DUAN H W,et al.Effects of different dietary lysine levels on performance of early-weaned piglets[J].Feed Industry,2010,31(20):5-7.(in Chinese)
[24]	EDMONDS M S,BAKER D H.Failure of excess dietary lysine to antagonize arginine in young pigs[J].J Nutr,1987, 117(8):1396-1401.
[25]	马文锋.猪肥育后期低氮日粮限制性氨基酸平衡模式的研究[D].北京:中国农业大学,2015.MA W F.Study on the limiting amino acid pattern for late finishing pigs fed protein reduced diet[D].Beijing:China Agricultural University,2015.(in Chinese)
[26]	袁中彪,李俊波,杨飞来,等.不同赖氨酸水平饲粮对断奶仔猪生长性能和营养物质消化率的影响[J].养猪,2008(6):3-4.YUAN Z B,LI J B,YANG F L,et al. Effect of the level of lysine on growth performances and nutrients digestibility in weaned piglets[J].Swine Production,2008(6):3-4.(in Chinese)
[27]	封飞飞,方伟,王淑楠,等.茶树油对育肥猪生长性能、器官指数、胴体性状和肉品质的影响[J].动物营养学报, 2017, 29(10):3620-3626.FENG F F,FANG W,WANG S N,et al.Effects of tea tree oil on growth performance,organ indexes,carcass characteristics and meat quality of finishing pigs[J].Chinese Journal of Animal Nutrition,2017,29(10):3620-3626.(in Chinese)
[28]	CHEN C Y,YANG B,ZENG Z J,et al.Genetic dissection of blood lipid traits by integrating genome-wide association study and gene expression profiling in a porcine model[J].BMC Genomics,2013,14(1):848.
[29]	ADARAMOYE O A,ADESANOYE O A,ADEWUMI O M,et al.Studies on the toxicological effect of nevirapine,an antiretroviral drug,on the liver,kidney and testis of male Wistar rats[J].Hum Exp Toxicol,2012,31(7):676-685.
[30]	SOUTHERN L L,BAKER D H.Performance and concentration of amino acids in plasma and urine of young pigs fed diets with excesses of either arginine or lysine[J].J Anim Sci,1982,55(4):857-866.
[31]	伍国耀,武振龙,戴兆来,等.猪对"非必需氨基酸"的营养需要[J].饲料工业,2013,34(16):60-64.WU G Y,WU Z L,DAI Z L,et al.Nutritional requirements for "non-essential amino acids" in pigs[J].Feed Industry,2013,34(16):60-64.(in Chinese)
[32]	KELLY D,SMYTH J A,MCCRACKEN K J.Digestive development of the early-weaned pig:1.Effect of continuous nutrient supply on the development of the digestive tract and on changes in digestive enzyme activity during the first week post-weaning[J].Br J Nutr,1991,65(2):169-180.
[33]	PLUSKE J R,THOMPSON M J,ATWOOD C S,et al.Maintenance of villus height and crypt depth,and enhancement of disaccharide digestion and monosaccharide absorption,in piglets fed on cows' whole milk after weaning[J].Br J Nutr,1996,76(3):409-422.
[34]	HE L Q,YANG H S,HOU Y Q,et al.Effects of dietary L-lysine intake on the intestinal mucosa and expression of CAT genes in weaned piglets[J].Amino Acids,2013,45(2):383-391.
[35]	施寿荣,刘勇强,张珊,等.赖氨酸和其他必需氨基酸对1~18日龄中速型黄羽肉鸡生长性能、血清生化指标、器官指数和肠道组织形态的影响[J].动物营养学报,2021,33(2):779-791.SHI S R,LIU Y Q,ZHANG S,et al.Effects of lysine and other essential amino acids on growth performance,serum biochemical indexes,organ indexes and intestinal morphology of medium-speed yellow-feather broilers during 1 to 18 days of age[J].Chinese Journal of Animal Nutrition,2021,33(2):779-791.(in Chinese)
[36]	SCHIERACK P,NORDHOFF M,POLLMANN M,et al.Characterization of a porcine intestinal epithelial cell line for in vitro studies of microbial pathogenesis in swine[J].Histochem Cell Biol,2006,125(3):293-305.
[37]	张海环.猪小肠赖氨酸转运体对氮源的响应规律及mTOR信号通路的调控研究[D].长春:吉林农业大学,2016.ZHANG H H.Study on lysine transports response law by different types of protein in the small intestines of weaned piglets and mTOR signaling pathway regulation in IEPC cells[D].Changchun:Jilin Agricultural University,2016.(in Chinese)
[38]	姜廷波.葡萄糖和赖氨酸水平对仔猪小肠上皮细胞赖氨酸吸收、转运的影响[D].长春:吉林农业大学,2017.JIANG Y B.Effects of glucose and lysine levels on the lysine absorption and transportation in intestinal epithelial cells of piglets[D].Changchun:Jilin Agricultural University,2017.(in Chinese)
[39]	王芳.赖氨酸蛋氨酸配比模式和葡萄糖水平影响酪蛋白合成关键基因表达[D].北京:中国农业科学院,2015.WANG F.Lysine/methionine pattern and glucose level regulate the expression of genes related to casein synthesis[D].Beijing:Chinese Academy of Agricultural Sciences,2015.(in Chinese)
[40]	WANG X,QIAO S Y,YIN Y L,et al.A deficiency or excess of dietary threonine reduces protein synthesis in jejunum and skeletal muscle of young pigs[J].J Nutr,2007,137(6):1442-1446.
[41]	WANG H,JI Y,WU G Y,et al.L-tryptophan activates mammalian target of rapamycin and enhances expression of tight junction proteins in intestinal porcine epithelial cells[J].J Nutr,2015,145(6):1156-1162.
[42]	高海娜,郑楠,胡菡,等.必需氨基酸通过哺乳动物雷帕霉素靶蛋白信号通路调控乳蛋白合成的研究进展[J].动物营养学报,2014,26(9):2451-2456.GAO H N,ZHENG N,HU H,et al.Research progress on the regulation of milk protein synthesis by essential amino acids through mammalian target of rapamycin signaling pathway[J].Chinese Journal of Animal Nutrition,2014,26(9):2451-2456.(in Chinese)
[43]	JI Y,WU Z L,DAI Z L,et al.Excessive L-cysteine induces vacuole-like cell death by activating endoplasmic reticulum stress and mitogen-activated protein kinase signaling in intestinal porcine epithelial cells[J].Amino Acids,2016,48(1):149-156.