家兔小肠不同区段营养物质转运载体相关基因的分布规律
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摘要
旨在研究家兔小肠不同区段营养物质转运载体相关基因的分布规律。本研究选取110日龄体重相近的健康白色獭兔10只,屠宰后采集十二指肠、空肠、回肠样品,采用real-time PCR研究家兔不同肠段小肽转运载体Pep T1,氨基酸转运载体CAT1、B~0AT、EAAT3、rBAT,葡萄糖转运载体SGLT1、GLUT2、GLUT5,以及脂肪酸转运载体FATP4的mRNA表达丰度。结果显示,小肽转运载体Pep T1 mRNA在十二指肠表达量最高,空肠略低;碱性氨基酸转运载体CAT1、兼性氨基酸转运载体rBAT和中性氨基酸转运载体B~0AT mRNA的表达量均在回肠最高,空肠次之;酸性氨基酸转运载体EAAT3 mRNA的表达量在空肠和回肠均较高;葡萄糖转运载体SGLT1和GLUT5 mRNA的表达量在十二指肠和空肠均较高;葡萄糖转运载体GLUT2和脂肪酸转运载体FATP4 mRNA的表达量则是空肠最高,十二指肠次之。结果表明,家兔肠道转运吸收小肽、葡萄糖和脂肪酸的主要部位是小肠前半段,转运吸收氨基酸的主要部位是小肠后半段。
The objective of this study was to investigate the distribution of nutrient transporter related genes in different segments of small intestine of rabbit. Ten healthy white rex rabbits at 110 days old with similar weight were chose to slaughter. The intestinal segment samples were collected from duodenum, jejunum, ileum for detecting the mRNA abundance of oligopeptide transporter Pep Tl, amino acid transporter CAT1, B~0AT, EAAT3, rBAT, glucose transporter SGLT1, GLUT2, GLUT5 and fatty acid transporter FATP4 by real-time PCR. The results showed that the abundance of oligopeptide transporter Pep T1 mRNA was the highest in duodenum, and lower in jejunum. The abundance of basic amino acid transporter CAT1, facultative amino acid transporter rBAT, and neutral amino acid transporter B~0AT mRNA were the highest in ileum, and lower in jejunum. The acidic amino acid transporter EAAT3 mRNA expression was higher in jejunum and ileum. The expression of glucose transporter SGLT1 and GLUT5 mRNA was higher in duodenum and jejunum. The abundance of glucose transporter GLUT2 and fatty acid transporter FATP4 mRNA were the highest in jejunum, and lower in duodenum. The result indicate that the main part in intestine for transporting and absorbing oligopeptide, glucose and fatty acid is the first half of the small intestine, the main part for transporting and absorbing amino acid is the posterior segment of the small intestine.
The objective of this study was to investigate the distribution of nutrient transporter related genes in different segments of small intestine of rabbit. Ten healthy white rex rabbits at 110 days old with similar weight were chose to slaughter. The intestinal segment samples were collected from duodenum, jejunum, ileum for detecting the mRNA abundance of oligopeptide transporter Pep Tl, amino acid transporter CAT1, B~0AT, EAAT3, rBAT, glucose transporter SGLT1, GLUT2, GLUT5 and fatty acid transporter FATP4 by real-time PCR. The results showed that the abundance of oligopeptide transporter Pep T1 mRNA was the highest in duodenum, and lower in jejunum. The abundance of basic amino acid transporter CAT1, facultative amino acid transporter rBAT, and neutral amino acid transporter B~0AT mRNA were the highest in ileum, and lower in jejunum. The acidic amino acid transporter EAAT3 mRNA expression was higher in jejunum and ileum. The expression of glucose transporter SGLT1 and GLUT5 mRNA was higher in duodenum and jejunum. The abundance of glucose transporter GLUT2 and fatty acid transporter FATP4 mRNA were the highest in jejunum, and lower in duodenum. The result indicate that the main part in intestine for transporting and absorbing oligopeptide, glucose and fatty acid is the first half of the small intestine, the main part for transporting and absorbing amino acid is the posterior segment of the small intestine.
引文
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[3] MISKA K B,FETTERER R H.Expression of amino acid and sugar transporters,aminopeptidase,and the di- and tri-peptide transporter PepT1;differences between modern fast growing broilers and broilers not selected for rapid growth[J].Poult Sci,2019,98(5):2272-2280.
[4] PONCET N,TAYLOR P M.The role of amino acid transporters in nutrition[J].Curr Opin Clin Nutr Metab Care,2013,16(1):57-65.
[5] SCALISE M,GALLUCCIO M,CONSOLE L,et al.The human SLC7A5 (LAT1):the intriguing histidine/large neutral amino acid transporter and its relevance to human health[J].Front Chem,2018,6:243.
[6] BR?ER S.Amino acid transport across mammalian intestinal and renal epithelia[J].Physiol Rev,2008,88(1):249-286.
[7] BYERS M S,HOWARD C,WANG X F.Avian and mammalian facilitative glucose transporters[J].Microarrays,2017,6(2):7.
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[9] KISHIDA K,MARTINEZ G,IIDA T,et al.D-allulose is a substrate of glucose transporter type 5 (GLUT5) in the small intestine[J].Food Chem,2019,277:604-608.
[10] LIN M H,MINER J H.Fatty acid transport protein 1 can compensate for fatty acid transport protein 4 in the developing mouse epidermis[J].J Invest Dermatol,2015,135(2):462-470.
[11] LENZ L S,MARX J,CHAMULITRAT W,et al.Adipocyte-specific inactivation of acyl-CoA synthetase fatty acid transport protein 4 (Fatp4) in mice causes adipose hypertrophy and alterations in metabolism of complex lipids under high fat diet[J].J Biol Chem,2011,286(41):35578-35587.
[12] 张爱华.肉仔鸡肠道主要营养素转运载体mRNA表达发育规律的研究[D].北京:中国农业科学院,2014.ZHANG A H.Ontogenetic regulation of nutrient transporters in the small intestine of broilers[D].Beijing:Chinese Academy of Agricultural Sciences,2014.(in Chinese)
[13] SPEIER J S,YADGARY L,UNI Z,et al.Gene expression of nutrient transporters and digestive enzymes in the yolk sac membrane and small intestine of the developing embryonic chick[J].Poult Sci,2012,91(8):1941-1949.
[14] 邹仕庚,冯定远,黄志毅,等.猪肠道寡肽转运载体1(PepT1)mRNA表达的肠段特异性和发育性变化[J].农业生物技术学报,2009,17(2):229-236.ZOU S G,FENG D Y,HUANG Z Y,et al.Segmental distribution and ontogenetic regulation of peptide transporter 1 (PepT1) mRNA expression in intestine of pigs[J].Journal of Agricultural Biotechnology,2009,17(2):229-236.(in Chinese)
[15] GILBERT E R,LI H,EMMERSON D A,et al.Developmental regulation of nutrient transporter and enzyme mRNA abundance in the small intestine of broilers[J].Poult Sci,2007,86(8):1739-1753.
[16] 吴丽娜.α-菠菜甾醇对小鼠小肠P糖蛋白和寡肽转运体PepT1药物转运体的影响研究[D].雅安:四川农业大学,2016.WU L N.The influence of α-spinasterol on the expression of P-glycoprotein and PepT1 on the small intestine of mice[D].Ya’an:Sichuan Agricultural University,2016.(in Chinese)
[17] 孙雅君,倪志鹤,陈玉林,等.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 Y J,NI Z H,CHEN Y L,et al.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.(in Chinese)
[18] FREEMAN T C,BENTSEN B S,THWAITES D T,et al.H+/Di-tripeptide transporter (PepT1) expression in the rabbit intestine[J].Pflügers Arch,1995,430(3):394-400.
[19] MUNCK L K,MUNCK B G.Variation in amino acid transport along the rabbit small intestine.Mutual jejunal carriers of leucine and lysine[J].Biochim Biophys Acta,1992,1116(2):83-90.
[20] 周响艳,左建军,职爱民,等.猪肠道碱性氨基酸转运载体(CAT1)mRNA表达的组织特异性和发育性变化[J].畜牧兽医学报,2008,39(2):170-175.ZHOU X Y,ZUO J J,ZHI A M,et al.Segmental distribution and ontogenetic regulation of cationic amino acid transporter mRNA expression in the small intestine of pigs[J].Acta Veterinaria et Zootechnica Sinica,2008,39(2):170-175.(in Chinese)
[21] 周英昊.陕北白绒山羊碱性氨基酸转运载体基因cDNA 克隆及时空表达分析[D].杨凌:西北农林科技大学,2015.ZHOU Y H.cDNA cloning and space-time expression analysis of cationic amino acid transporters gene in Shanbei White Cashmere goat[D].Yangling:Northwest A&F University,2015.(in Chinese)
[22] DAVE M H,SCHULZ N,ZECEVIC M,et al.Expression of heteromeric amino acid transporters along the murine intestine[J].J Physiol,2004,558(2):597-610.
[23] ROMEO E,DAVE M H,BACIC D,et al.Luminal kidney and intestine SLC6 amino acid transporters of B0AT-cluster and their tissue distribution in Mus musculus[J].Am J Physiol Renal,2006,290(2):376-383.
[24] 王修启,谭会泽,束刚,等.鸡肠道SGLT1和GLUT2 mRNA表达的组织特异性研究[J].畜牧兽医学报,2006,37(1):12-17.WANG X Q,TAN H Z,SHU G,et al.Study on tissue-specific expression of SGLT1 and GLUT2 mRNA in chicken intestine[J].Acta Veterinaria et Zootechnica Sinica,2006,37(1):12-17.(in Chinese)
[25] 董信阳.鸽早期小肠发育及碳水化合物对其调控的研究[D].杭州:浙江大学,2013.DONG X Y.Research on early development of the pigeon small intestine and its regulation by carbohydrates[D].Hangzhou:Zhejiang University,2013.(in Chinese)
[2] 许庆彪.奶牛瓣胃上皮细胞对小肽的吸收机制及小肽转运载体1的特征研究[D].杭州:浙江大学,2015.XU Q B.The mechanism of small peptide absorption in bovine omasal epithelial cells and characterization of bovine peptide transporter 1[D].Hangzhou:Zhejiang University,2015.(in Chinese)
[3] MISKA K B,FETTERER R H.Expression of amino acid and sugar transporters,aminopeptidase,and the di- and tri-peptide transporter PepT1;differences between modern fast growing broilers and broilers not selected for rapid growth[J].Poult Sci,2019,98(5):2272-2280.
[4] PONCET N,TAYLOR P M.The role of amino acid transporters in nutrition[J].Curr Opin Clin Nutr Metab Care,2013,16(1):57-65.
[5] SCALISE M,GALLUCCIO M,CONSOLE L,et al.The human SLC7A5 (LAT1):the intriguing histidine/large neutral amino acid transporter and its relevance to human health[J].Front Chem,2018,6:243.
[6] BR?ER S.Amino acid transport across mammalian intestinal and renal epithelia[J].Physiol Rev,2008,88(1):249-286.
[7] BYERS M S,HOWARD C,WANG X F.Avian and mammalian facilitative glucose transporters[J].Microarrays,2017,6(2):7.
[8] HUMMEL C S,LU C,LOO D D F,et al.Glucose transport by human renal Na+/D-glucose cotransporters SGLT1 and SGLT2[J].Am J Physiol Cell Physiol,2011,300(1):C14-C21.
[9] KISHIDA K,MARTINEZ G,IIDA T,et al.D-allulose is a substrate of glucose transporter type 5 (GLUT5) in the small intestine[J].Food Chem,2019,277:604-608.
[10] LIN M H,MINER J H.Fatty acid transport protein 1 can compensate for fatty acid transport protein 4 in the developing mouse epidermis[J].J Invest Dermatol,2015,135(2):462-470.
[11] LENZ L S,MARX J,CHAMULITRAT W,et al.Adipocyte-specific inactivation of acyl-CoA synthetase fatty acid transport protein 4 (Fatp4) in mice causes adipose hypertrophy and alterations in metabolism of complex lipids under high fat diet[J].J Biol Chem,2011,286(41):35578-35587.
[12] 张爱华.肉仔鸡肠道主要营养素转运载体mRNA表达发育规律的研究[D].北京:中国农业科学院,2014.ZHANG A H.Ontogenetic regulation of nutrient transporters in the small intestine of broilers[D].Beijing:Chinese Academy of Agricultural Sciences,2014.(in Chinese)
[13] SPEIER J S,YADGARY L,UNI Z,et al.Gene expression of nutrient transporters and digestive enzymes in the yolk sac membrane and small intestine of the developing embryonic chick[J].Poult Sci,2012,91(8):1941-1949.
[14] 邹仕庚,冯定远,黄志毅,等.猪肠道寡肽转运载体1(PepT1)mRNA表达的肠段特异性和发育性变化[J].农业生物技术学报,2009,17(2):229-236.ZOU S G,FENG D Y,HUANG Z Y,et al.Segmental distribution and ontogenetic regulation of peptide transporter 1 (PepT1) mRNA expression in intestine of pigs[J].Journal of Agricultural Biotechnology,2009,17(2):229-236.(in Chinese)
[15] GILBERT E R,LI H,EMMERSON D A,et al.Developmental regulation of nutrient transporter and enzyme mRNA abundance in the small intestine of broilers[J].Poult Sci,2007,86(8):1739-1753.
[16] 吴丽娜.α-菠菜甾醇对小鼠小肠P糖蛋白和寡肽转运体PepT1药物转运体的影响研究[D].雅安:四川农业大学,2016.WU L N.The influence of α-spinasterol on the expression of P-glycoprotein and PepT1 on the small intestine of mice[D].Ya’an:Sichuan Agricultural University,2016.(in Chinese)
[17] 孙雅君,倪志鹤,陈玉林,等.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 Y J,NI Z H,CHEN Y L,et al.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.(in Chinese)
[18] FREEMAN T C,BENTSEN B S,THWAITES D T,et al.H+/Di-tripeptide transporter (PepT1) expression in the rabbit intestine[J].Pflügers Arch,1995,430(3):394-400.
[19] MUNCK L K,MUNCK B G.Variation in amino acid transport along the rabbit small intestine.Mutual jejunal carriers of leucine and lysine[J].Biochim Biophys Acta,1992,1116(2):83-90.
[20] 周响艳,左建军,职爱民,等.猪肠道碱性氨基酸转运载体(CAT1)mRNA表达的组织特异性和发育性变化[J].畜牧兽医学报,2008,39(2):170-175.ZHOU X Y,ZUO J J,ZHI A M,et al.Segmental distribution and ontogenetic regulation of cationic amino acid transporter mRNA expression in the small intestine of pigs[J].Acta Veterinaria et Zootechnica Sinica,2008,39(2):170-175.(in Chinese)
[21] 周英昊.陕北白绒山羊碱性氨基酸转运载体基因cDNA 克隆及时空表达分析[D].杨凌:西北农林科技大学,2015.ZHOU Y H.cDNA cloning and space-time expression analysis of cationic amino acid transporters gene in Shanbei White Cashmere goat[D].Yangling:Northwest A&F University,2015.(in Chinese)
[22] DAVE M H,SCHULZ N,ZECEVIC M,et al.Expression of heteromeric amino acid transporters along the murine intestine[J].J Physiol,2004,558(2):597-610.
[23] ROMEO E,DAVE M H,BACIC D,et al.Luminal kidney and intestine SLC6 amino acid transporters of B0AT-cluster and their tissue distribution in Mus musculus[J].Am J Physiol Renal,2006,290(2):376-383.
[24] 王修启,谭会泽,束刚,等.鸡肠道SGLT1和GLUT2 mRNA表达的组织特异性研究[J].畜牧兽医学报,2006,37(1):12-17.WANG X Q,TAN H Z,SHU G,et al.Study on tissue-specific expression of SGLT1 and GLUT2 mRNA in chicken intestine[J].Acta Veterinaria et Zootechnica Sinica,2006,37(1):12-17.(in Chinese)
[25] 董信阳.鸽早期小肠发育及碳水化合物对其调控的研究[D].杭州:浙江大学,2013.DONG X Y.Research on early development of the pigeon small intestine and its regulation by carbohydrates[D].Hangzhou:Zhejiang University,2013.(in Chinese)
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