绵羊胃肠道游离氨基酸与小肽分布及其转运载体相关基因表达的研究

doi:10.11843/j.issn.0366-6964.2017.03.009

摘要/Abstract

摘要：

旨在研究绵羊胃肠道内游离氨基酸（FAA）和小肽（PAA）的数量分布和碱性氨基酸转运载体CAT1、酸性氨基酸转运载体EAAT3、中性氨基酸转运载体y⁺LAT2和ASCT2、小肽转运载体PepT1、氨基肽酶APN、二肽酶DPEP2 mRNA在胃肠道的表达规律。试验选取18月龄健康小尾寒羊6只，屠宰后收集瘤胃、十二指肠、空肠、回肠和盲肠的食糜及对应肠道组织，对食糜上清液的小肽和游离氨基酸进行测定，并用实时定量PCR对基因表达量进行测定。结果显示：1）绵羊食糜多数游离氨基酸浓度从高到低的分布为空肠、十二指肠、回肠、瘤胃和盲肠，前二者显著高于其他部位（P<0.05）；不同部位游离氨基酸组成比例不同；多数小肽浓度从高到低的分布为十二指肠、空肠、回肠、盲肠和瘤胃，除瘤胃和盲肠外各部位间差异显著（P<0.05）；不同部位小肽的组成比例不同；小肽占总氨基酸（TAA）的比例在空肠显著低于其他部位（P<0.05）。2）CAT1 mRNA在空肠表达量最高，但不同部位间差异不显著；EAAT3、y⁺LAT2、PepT1、APN mRNA表达规律相似，均在回肠表达量最高；ASCT2 mRNA在十二指肠表达量最高，DPEP2 mRNA在盲肠表达量最高。结果提示，胃肠道不同区段游离氨基酸和小肽的浓度和组成不同；游离氨基酸的主要释放部位是空肠，主要吸收部位是回肠；小肽的主要释放部位是十二指肠，主要消化部位是空肠，在肠系膜系统的主要吸收部位是回肠；瘤胃和小肠各区段吸收碱性氨基酸的能力基本相同；酸性和多数中性氨基酸主要吸收部位是回肠，少数中性氨基酸主要吸收部位是空肠。本研究为绵羊小肠氨基酸营养和蛋白质消化吸收规律的研究提供了理论依据。

关键词: 绵羊, 小肽, 游离氨基酸, 转运载体, mRNA表达

Abstract:

This study aimed to clarify quantitative distributions of free amino acids (FAA) and small peptides (PAA) in gastrointestinal tract of sheep and the mRNA expression of CAT1 (cationic amino acid transporter), EAAT3 (acidic amino acid transporter), y⁺LAT2, ASCT2(neutral amino acid transporters), PepT1 (small peptide transporter), APN (aminopeptidase) and DPEP2 (dipeptidase) in gastrointestinal tract of sheep. A total of 6 healthy Small-tail Han sheep at the age of 18 months were slaughtered to collect chyme and tissues of rumen, duodenum, jejunum, ileum and cecum. Then concentrations of the most of FAA and PAA in supernatant of chyme were detected and the mRNA expression of related genes were quantified by RT-PCR. The results showed that: 1) The concentrations of the most of FAA from high to low was jejunum, duodenum, ileum, rumen, cecum. The concentrations of FAA in jejunum and duodenum were significant higher than that in other parts(P<0.05). Proportions of FAA in these parts were different. The concentrations of the most of PAA from high to low was duodenum, jejunum, ileum, cecum, rumen. Except for cecum and rumen, they were significant different from each other(P<0.05). Proportions of PAA in these parts were different. The proportion of PAA in total amino acids was the significantly lowest in jejunum(P<0.05). 2) CAT1 mRNA in jejunum had the highest abundance, but there were no significant difference among 5 parts. EAAT3, y⁺LAT2, PepT1 and APN mRNA had a similar expression pattern that they all had the highest abundance in ileum. ASCT2 mRNA had the highest abundance in duodenum, and DPEP2 mRNA had the highest abundance in cecum. Results indicate that concentrations and proportions of FAA in these parts are different, the same as PAA. The major secretion site of FAA is jejunum. The major absorption site of FAA is ileum. The major production site of PAA is duodenum. The major digestion site of FAA is jejunum. The major absorption site of FAA in mesenteric system is ileum. The absorbility of cationic amino acids in 5 parts are similar. Acidic amino acids and the most of neutral amino acids are absorbed major in ileum, but some of neutral amino acids are absorbed major in jejunum. This study provides the theory basis for amino acid nutrition in small intestine of sheep and for digestion and absorption regular pattern of protein of sheep.

Key words: sheep, small peptides, free amino acids, transporter, mRNA expression

中图分类号:

S826
S815

孙雅君，倪志鹤，陈玉林，张恩平，王永军. 绵羊胃肠道游离氨基酸与小肽分布及其转运载体相关基因表达的研究[J]. 畜牧兽医学报, 2017, 48(3): 462-473.

SUN Ya-jun, NI Zhi-he, CHEN Yu-lin, ZHANG En-ping, WANG Yong-jun. Study on Distribution of Free Amino Acid and Small Peptide and the Expression of Related Transporter Genes in Gastrointestinal Tract of Sheep[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2017, 48(3): 462-473.

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