复合氨基酸对断奶仔猪肝脏能量状态、游离氨基酸含量及其相关基因表达的影响

doi:10.16431/j.cnki.1671-7236.2020.08.012

摘要/Abstract

摘要： 为进一步探明复合功能性氨基酸的应用效果与机理，在前期研究的基础上，本试验研究了日粮中添加复合氨基酸对断奶仔猪肝脏能量状态、游离氨基酸及相关基因表达的影响。将16头体重相近的（21±3）日龄断奶仔猪随机分为对照组和复合氨基酸组，每组8个重复。分别饲喂添加0.99%丙氨酸（与试验组等氮）和1.00%复合氨基酸（谷氨酸：谷氨酰胺：精氨酸：甘氨酸：N-乙酰半胱氨酸=5：2：2：1：0.5）的基础日粮。于试验第21天前腔静脉采血，屠宰取肝脏样品，待测。结果表明，与对照组相比，复合氨基酸显著降低了仔猪血浆ALT和AST活性（P<0.05）；显著提高了仔猪肝脏总蛋白含量、RNA/DNA和总蛋白/DNA（P<0.05），显著降低了仔猪肝脏MDA和H₂O₂含量（P<0.05）；显著提高了肝脏精氨酸和二磷酸腺苷（ADP）含量（P<0.05）；显著上调了肝脏氨基酸转运载体b^0，+AT和y⁺LAT2基因、谷胱甘肽硫转移酶ω2基因（GSTO2）、干扰素γ基因（IFN-γ）、脂蛋白酯酶基因（LPL）的表达（P<0.05），显著下调了脂肪酸合成酶基因（FASN）的表达（P<0.05）。以上结果表明，日粮中添加1.00%复合氨基酸可以改善肝功能，提高抗氧化能力，可以调控仔猪肝脏能量状态及游离氨基酸代谢，并可能影响肝脏的脂肪代谢和免疫功能。

关键词: 复合氨基酸; 断奶仔猪; 肝脏; 能量状态; 游离氨基酸; 基因

Abstract: In order to further explore the application effects and mechanism of an amino acid blend (AAB),this study investigated the effects of dietary supplementation with AAB on liver energy state,free amino acids and relative expression of related genes in weaned piglets following our previous studies.Sixteen (21±3) days old weaned piglets with similar body weight were randomly assigned to control group and AAB group,with 8 replicates per group.The diet of the control group was basal diet supplemented with 0.99% alanine (isonitrogenous control),the diet of AAB group was basal diet supplemented with 1.00% AAB (glutamate:glutamine:glycine:arginine:N-acetylcysteine=5:2:2:1:0.5).On day 21 of the trial,the blood and liver samples were obtained from all piglets.The results indicated that:Compared to the control group,AAB group decreased plasma ALT and AST activity in piglets (P<0.05),increased the total protein content,RNA/DNA and total protein/DNA in the liver of piglets(P<0.05),reduced the MDA and H₂O₂ contents in the liver of piglets (P<0.05),increased the liver arginine and adenosine diphosphate (ADP) levels (P<0.05),increased mRNA levels of amino acid transporter b^0,+ AT and y⁺ LAT2 genes,glutathione S-transferase ω2 gene (GSTO2),interferon-γ gene (IFN-γ),lipoprotein lipase gene (LPL) (P<0.05),decreased mRNA levels of fatty acid synthase gene (FASN) (P<0.05).The above results showed that the addition of 1.00% AAB could improve the liver function,improve the antioxidant capacity,regulate the energy state and free amino acid metabolism of piglet liver,and might affect the fat metabolism and immune function of liver.

Key words: amino acid blend; weaned piglets; liver; energy state; free amino acids; gene

中图分类号:

S828.5

董毅, 王倩, 孟禹璇, 朱哲坤, 田俊杰, 王蕾, 赵迪, 侯永清. 复合氨基酸对断奶仔猪肝脏能量状态、游离氨基酸含量及其相关基因表达的影响[J]. 中国畜牧兽医, 2020, 47(8): 2445-2453.

DONG Yi, WANG Qian, MENG Yuxuan, ZHU Zhekun, TIAN Junjie, WANG Lei, ZHAO Di, HOU Yongqing. Effects of Amino Acid Blend on Liver Energy State,Free Amino Acids and Relative Expression of Related Genes in Weaned Piglets[J]. China Animal Husbandry and Veterinary Medicine, 2020, 47(8): 2445-2453.

参考文献

[1] 杨震国,张伟,侯永清,等.N-乙酰半胱氨酸对脂多糖刺激仔猪免疫应激和能量状况的影响[J].畜牧兽医学报,2012,43(4):564-571. YANG Z G,ZHANG W,HOU Y Q,et al.Effects of N-acetylcysteine on immunological stress and energy status of piglets chronically challenged with lipopolysaccharide[J].Acta Veterinaria et Zootechnica Sinica,2012,43(4):564-571.(in Chinese)
[2] 汪龙梅,王秀英,吴欢听,等.谷氨酸对仔猪肝脏能量代谢及关键酶和相关调节因子mRNA表达的影响[J].中国畜牧杂志,2016,52(23):38-43. WANG L M,WANG X Y,WU H T,et al.Effect of glutamate on energy metabolism and the expression of mRNA of key enzymes and related regulatory factors in the liver of piglets challenged with lipopolysaccharide[J].Chinese Journal of Animal Science,2016,52(23):38-43.(in Chinese)
[3] WU G Y,BAZER F W,DAI Z,et al.Amino acid nutrition in animals:Protein synthesis and beyond[J].Annual Review of Animal Biosciences,2014,2(1):387-417.
[4] WU G Y.Functional amino acids in growth,reproduction,and health[J].Advances in Nutrition:An International Review Journal,2010,1(1):31-37.
[5] HOU Y Q,WANG L,YI D,et al.N-acetylcysteine and intestinal health:A focus on mechanisms of its actions[J].Frontiers in Bioscience (Landmark Ed),2015,20(5):872-891.
[6] JIAO N,WU Z,JI Y,et al.L-glutamate enhances barrier and antioxidative functions in intestinal porcine epithelial cells[J].Journal of Nutrition,2015,145(10):2258-2264.
[7] WANG W,WU Z,LIN G,et al.Glycine stimulates protein synthesis and inhibits oxidative stress in pig small intestinal epithelial cells[J].Journal of Nutrition,2014,144(10):1540-1548.
[8] 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.
[9] YI D,HOU Y Q,WANG L,et al.L-glutamine enhances enterocyte growth via activation of the mTOR signaling pathway independently of AMPK[J].Amino Acids,2015,47(1):65-78.
[10] YI D,HOU Y Q,WANG L,et al.N-acetylcysteine stimulates protein synthesis in enterocytes independently of glutathione synthesis[J].Amino Acids,2016,48(2):523-533.
[11] DUAN J,YIN J,WU M,et al.Dietary glutamate supplementation ameliorates mycotoxin-induced abnormalities in the intestinal structure and expression of amino acid transporters in young pigs[J].PLoS One,2013,9(11):e112357.
[12] HU C J,JIANG Q Y,ZHANG T,et al.Dietary supplementation with arginine and glutamic acid modifes growth performance,carcass traits,and meat quality in growing-fnishing pigs[J].Journal of Animal Science,2017,95(6):1680-2689.
[13] YI D,LI B C,HOU Y Q,et al.Dietary supplementation with an amino acid blend enhances intestinal function in piglets[J].Amino Acids,2018,50(8):1089-1100.
[14] JOHNSON L R,CHANDLER A M.RNA and DNA of gastric and duodenal mucosa in antrectomized and gastrin-treated rats[J].American Journal of Physiology-Legacy Content,1973,224(4):937-940.
[15] FU W J,STROMBERG A J,VIELE K,et al.Statistics and bioinformatics in nutritional sciences:Analysis of complex data in the era of systems biology[J].Journal of Nutritional Biochemistry,2009,21(7):561-572.
[16] CHEN F,LIU Y L,ZHU H L,et al.Fish oil attenuates liver injury caused by LPS in weaned pigs associated with inhibition of TLR4 and nucleotide-binding oligomerization domain protein signaling pathways[J].Innate Immunity,2013,19(5):504-515.
[17] LI Q,LIU Y,CHE Z,et al.Dietary L-arginine supplementation alleviates liver injury caused by Escherichia coli LPS in weaned pigs[J].Innate Immunity,2012,18(6):804-814.
[18] NAKAO A,TAKI S,YASUI M,et al.The fate of intravenously injected endotoxin in normal rats and in rats with liver failure[J].Hepatology,1994,19(5):1251-1256.
[19] HE P,NODA Y,SUGIYAMA K.Green tea suppresses lipopolysaccharide-induced liver injury in D-galactosamine sensitized rats[J].Journal of Nutrition,2001,131(5):1560-1567.
[20] ZHAO D,WU T,YI D,et al.Dietary supplementation with lactobacillus casei alleviates lipopolysaccharide-induced liver injury in a porcine model[J].International Journal of Molecular Sciences,2017,18(12):2535-2546.
[21] YI D,HOU Y Q,WANG L,et al.Dietary N-acetylcysteine supplementation alleviates liver injury in lipopolysaccharide-challenged piglets[J].British Journal of Nutrition,2014,111(1):46-54.
[22] WU L,WANG W,YAO K,et al.Effects of dietary arginine and glutamine on alleviating the impairment induced by deoxynivalenol stress and immune relevant cytokines in growing pigs[J].PLoS One,2013,8(7):e69502.
[23] LIU Y,CHEN F,ODLE J,et al.Fish oil increases muscle protein mass and modulates Akt/FOXO,TLR4,and NOD signaling in weanling piglets after lipopolysaccharide challenge[J].Journal of Nutrition,2013,143(8):1331-1339.
[24] 付大波,王友炜,侯永清,等.α-酮戊二酸对脂多糖刺激断奶仔猪肌肉能量代谢的影响[J].中国农业科学,2011,44(4):814-822. FU D B,WANG Y W,HOU Y Q,et al.Effects of α-ketoglutarate on muscle energy metabolism in weanling pigs chronically challenged with lipopolysaccharide[J].Scientia Agricultura Sinence,2011,44(4):814-822.(in Chinese)
[25] PIA D,LIUA Y,SHIA H,et al.Dietary supplementation of aspartate enhances intestinal integrity and energy status in weanling piglets after lipopolysaccharide challenge[J].The Journal of Nutritional Biochemistry,2014,25(4):456-462.
[26] KANG P,LIU Y L,ZHU H L,et al.The effect of aspartate on the energy metabolism in the liver of weanling pigs challenged with lipopolysaccharide[J].European Journal of Nutrition,2014,54(4):581-588.
[27] 张琳,王秀英,吴欢听,等.甘氨酸对脂多糖刺激仔猪肝脏能量代谢及相关基因表达的调控作用[J].动物营养学报,2016,28(12):4006-4013. ZHANG L,WANG X Y,WU H T,et al.Regulatory role of glycine on energy metabolism and its related genes' expression in liver of piglets after lipopolysaccharide challenge[J].Chinese Journal of Animal Nutrition,2016,28(12):4006-4013.(in Chinese)
[28] STRANGE R C,SPITERI M A,RAMACHANDRAN S,et al.Glutathione-S-transferase family of enzymes[J].Mutation Research,2001,482(1-2):21-26.
[29] 杜纪坤,黄青阳.脂蛋白脂酶基因的研究进展[J].遗传,2007,29(1):8-16. DU J K,HUANG Q Y.Research progress of lipoprotein lipase gene[J].Hereditas,2007,29(1):8-16.(in Chinese)
[30] 周国利,金海国.脂肪酸合成酶基因(FASN)的研究进展[J].安徽农业科学,2008,36(9):3559-3560. ZHOU G L,JIN H G.Research progress in the fatty acid synthase gene[J].Journal of Anhui Agricultural Science,2008,36(9):3559-3560.(in Chinese)
[31] 黄思琪,曲红焱,黄大鹏,等.L-精氨酸对冷应激仔猪生长性能、免疫功能及肝脏、肾脏中肿瘤坏死因子-α、干扰素-γ基因表达量的影响[J].动物营养学报,2019,31(1):131-139. HUANG S Q,QU H Y,HUANG D P,et al.Effects of L-arginine on growth performance,immune function and genes expression levels of tumor necrosis factor-α and interferon-γ in liver and kidney of cold-stressed piglets[J].Chinese Journal of Animal Nutrition,2019,31(1):131-139.(in Chinese)
[32] WU G Y,BAZER F W,DAVIS T A,et al.Arginine metabolism and nutrition in growth,health and disease[J].Amino Acids,2009,37(1):153-168.