呼伦贝尔羊重要脂肪相关基因mRNA差异表达分析

doi:10.11843/j.issn.0366-6964.2019.09.022

摘要/Abstract

摘要： 旨在基于前期尾部极端大小呼伦贝尔羊的尾部脂肪转录组测序研究结果，选择10个重要的脂肪相关基因，研究其在不同组织中的表达情况，进一步验证这些基因与绵羊脂肪代谢的关系。本研究利用实时荧光定量方法检测CFD、PLIN4、THY1、IL-18、PTPN11、LPL、IRX3、HOXA10、MID1IP1、UGCG基因在6月龄呼伦贝尔羊大尾和小尾品系7种组织（尾部脂肪、臀部脂肪、皮下脂肪、肾周脂肪、网膜脂肪、肝、肌肉）中mRNA的表达差异。研究结果表明：1）10个基因除了在肌肉中存在痕量表达外，在其他各个组织均有表达，而且在不同品系间的一个或多个组织中的表达量存在显著差异。2）IRX3和UGCG基因在大尾羊品系尾部脂肪组织中的表达量均极显著低于小尾羊品系（P<0.01），表明这两个基因与绵羊尾部脂肪代谢有直接关系。3）PLIN4和PTPN11基因在各个组织的表达趋势相近，且在大尾羊品系的皮下及网膜脂肪组织的表达量极显著高于小尾羊品系（P<0.01），而THY1基因与它们的表达趋势正好相反，在大尾品系皮下及网膜脂肪的表达量显著低于小尾羊品系（分别为P<0.01，P<0.05）。4）LPL基因只在小尾羊的臀部脂肪组织的表达量极显著高于大尾羊（P<0.01）；仅有CFD基因在肝中高表达，并在大尾羊中的表达量极显著高于小尾羊（P<0.01），而其他基因在肝中低表达或者痕量表达；HOXA10基因仅在肾周脂肪组织中高表达，并且在大尾羊中的表达量极显著高于小尾羊（P<0.01）。研究结果可为阐明绵羊脂肪代谢的分子调控机理提供参考。

Abstract: Based on the transcriptome sequencing results of tail adipose tissue between the big-tailed and small-tailed Hulun Buir sheep, 10 important genes related to fat were selected to explore their expression in different tissues and investigate the relationship between genes and fat metabolism in sheep. The mRNA expression difference of candidate genes, including CFD, PLIN4, THY1, IL-18, PTPN11, LPL, IRX3, HOXA10, MID1IP1 and UGCG, were detected in 7 tissues (tail fat, hip fat, subcutaneous fat, perirenal fat, omental fat, liver and muscle) between big-tailed and small-tailed Hulun Buir sheep of 6-month-old by real-time fluorescence quantitative method. The results showed that:1) Ten genes were expressed in other tissues except for trace expression in muscle, and there was a significant difference at the expression levels in one or more tissues between different lines. 2) The expression levels of IRX3 and UGCG genes in the tail adipose tissue of the big-tailed sheep were significantly lower than those in the small-tailed sheep (P<0.01), indicating that these 2 genes were directly related to fat metabolism in the tail of sheep. 3) The expression trends of PLIN4 and PTPN11 genes in different tissues were similar, and the expression levels in subcutaneous and omental adipose tissues in the big-tailed sheep were higher than those in the small-tailed sheep (P<0.01), while the expression trend of THY1 gene was opposite, the expression in subcutaneous and omental fat tissues in the big-tailed sheep was lower than that in the small-tailed sheep (P<0.01, P<0.05). 4) The expression of LPL gene in the hip adipose tissue of the small-tailed sheep was significantly higher than that of the big-tailed sheep (P<0.01); only the CFD gene was highly expressed in the liver and higher in the big-tailed sheep than that in the small-tailed sheep (P<0.01), while other genes were lowly or tracely expressed in the liver; HOXA10 gene was highly expressed only in perirenal adipose tissue, and the expression level in big-tailed sheep was higher than that in small-tailed sheep (P<0.01). The results will provide a reference for further elucidate the molecular mechanism of fat metabolism in sheep.

中图分类号:

S826.2

邓天宇, 樊红樱, 杜立新, 赵福平, 王立贤. 呼伦贝尔羊重要脂肪相关基因mRNA差异表达分析[J]. 畜牧兽医学报, 2019, 50(9): 1926-1935.

DENG Tianyu, FAN Hongying, DU Lixin, ZHAO Fuping, WANG Lixian. Differential Expression Analysis of Important Candidate Genes mRNA Related to Fat in Hulun Buir Sheep[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2019, 50(9): 1926-1935.

0
/ / 推荐

导出引用管理器 EndNote|Reference Manager|ProCite|BibTeX|RefWorks

链接本文: https://www.xmsyxb.com/CN/10.11843/j.issn.0366-6964.2019.09.022

https://www.xmsyxb.com/CN/Y2019/V50/I9/1926

参考文献

[1] 司智陟.基于营养目标的我国肉类供需分析[D].北京:中国农业科学院,2012.
SI Z Z.Study on meat supply and demand with the goal of nutrition in China[D].Beijing:Chinese Academy of Agricultural Sciences,2012.(in Chinese)
[2] FARAHANI A H K,SHAHRBABAK H M,SHAHRBABAK M M,et al.Relationship of fat-tail and body measurements with some economic important traits in fat-tail Makoei breed of Iranian sheep[J].Afr J Biotechnol,2010,9(36):5989-5992.
[3] KHALDARI M,KASHAN N E J,AFZALZADEH A,et al.Growth and carcass characteristics of crossbred progeny from lean-tailed and fat-tailed sheep breeds[J].South Aft J Anim Sci,2007,37(1):51-56.
[4] MORADI M H,NEJATI-JAVAREMI A,MORADI-SHAHRBABAK M,et al.Genomic scan of selective sweeps in thin and fat tail sheep breeds for identifying of candidate regions associated with fat deposition[J].BMC Genet,2012,13:10.
[5] 甘尚权,沈敏,李欢,等.X染色体60149273位点在脂尾(臀)和瘦尾绵羊品种中的多态性及其基因定位[J].中国农业科学,2013,46(22):4791-4799.
GAN S Q,SHEN M,LI H,et al.Polymorphism of the 60149273th Loci on X chromosome among fat tail and thin tail breeds and its gene mapping[J].Scientia Agricultura Sinica,2013,46(22):4791-4799.(in Chinese)
[6] 董淑霞,康静,王永军,等.草原优良新品种——呼伦贝尔羊简介[J].畜牧兽医科技信息,2010(3):104-105.
DONG S X,KANG J,WANG Y J,et al.A new variety of grassland——introduction to Hulunbeier sheep[J].Chinese Journal of Animal Husbandry and Veterinary Medicine,2010(3):104-105.(in Chinese)
[7] 樊红樱.呼伦贝尔绵羊尾部脂肪组织的转录组差异表达分析[D].兰州:甘肃农业大学,2016.
FAN H Y.Transcriptomic difference analysis for tail adipose tissue of Hulun Buir sheep[D].Lanzhou:Gansu Agricultural University, 2016.(in Chinese)
[8] SMEMO S,TENA J J,KIM K H,et al.Obesity-associated variants within FTO form long-range functional connections with IRX3[J]. Nature,2014,507(7492):371-375.
[9] CLAUSSNITZER M,DANKEL S N,KIM K H,et al.FTO obesity variant circuitry and adipocyte browning in humans[J].N Engl J Med,2015,373(10):895-907.
[10] KANG D J,ZHOU G X,ZHOU S W,et al.Comparative transcriptome analysis reveals potentially novel roles of Homeobox genes in adipose deposition in fat-tailed sheep[J].Sci Rep,2017,7(1):14491.
[11] 宁越,吴森,张乐,等.IRX3基因在秦川牛不同组织中的表达分析[J].西北农林科技大学学报:自然科学版, 2018, 46(6):15-19,47.
NING Y,WU S,ZHANG L,et al.Expression of IRX3 gene in different tissues of Qinchuan cattle[J].Journal of Northwest A&F University:Natural Science Edition,2018,46(6):15-19,47.(in Chinese)
[12] NOMURA K H,MURATA D,HAYASHI Y,et al.Ceramide glucosyltransferase of the nematode Caenorhabditis elegans is involved in oocyte formation and in early embryonic cell division[J].Glycobiology,2011,21(6):834-848.
[13] WEGNER M S,GRUBER L,MATTJUS P,et al.The UDP-glucose ceramide glycosyltransferase (UGCG) and the link to multidrug resistance protein 1(MDR1)[J].BMC Cancer,2018,18(1):153.
[14] HAYASHI Y,NEMOTO-SASAKI Y,MATSUMOTO N,et al.Complex formation of sphingomyelin synthase 1 with glucosylceramide synthase increases sphingomyelin and decreases glucosylceramide levels[J].J Biol Chem,2018, 293(45):17505-17522.
[15] NORDSTRÖM V,WILLERSHÄUSER M,HERZER S,et al.Neuronal expression of glucosylceramide synthase in central nervous system regulates body weight and energy homeostasis[J].PLoS Biol,2013,11(3):e1001506.
[16] 韦璇,徐小春,杨雨鑫,等.CCAAT/增强子结合蛋白α(C/EBPα)与脂蛋白酯酶(LPL)基因在不同尾型绵羊品种尾部脂肪组织中的表达[J].农业生物技术学报,2014,22(5):598-606.
WEI X,XU X C,YANG Y X,et al.Differential expression of CCAAT/enhancer binding protein α(C/EBPα) and Lipoprotein Lipase(LPL) genes in tail adipose tissues of sheep (Ovis aries) with different types of tail[J].Journal of Agricultural Biotechnology, 2014, 22(5):598-606.(in Chinese)
[17] REN M Q,WEGNER J,BELLMANN O,et al.Comparing mRNA levels of genes encoding leptin,leptin receptor,and lipoprotein lipase between dairy and beef cattle[J].Domest Anim Endocrinol,2002,23(3):371-381.
[18] BARBER M C,WARD R J,RICHARDS S E,et al.Ovine adipose tissue monounsaturated fat content is correlated to depot-specific expression of the stearoyl-CoA desaturase gene[J].J Anim Sci,2000,78(1):62-68.
[19] ROPKA-MOLIK K,KNAPIK J,PIESZKA M,et al.Nutritional modification of SCD,ACACA and LPL gene expressions in different ovine tissues[J].Arch Anim Breed,2017,60(3):243-250.
[20] COLEMAN D N,MURPHY K D,RELLING A E.Prepartum fatty acid supplementation in sheep.Ⅱ.Supplementation of eicosapentaenoic acid and docosahexaenoic acid during late gestation alters the fatty acid profile of plasma,colostrum,milk and adipose tissue,and increases lipogenic gene expression of adipose tissue[J].J Anim Sci,2018,96(3):1181-1204.
[21] BRASAEMLE D L.Thematic review series:Adipocyte biology.The perilipin family of structural lipid droplet proteins:stabilization of lipid droplets and control of lipolysis[J].J Lipid Res,2007,48(12):2547-2559.
[22] DE LA ROSA RODRIGUEZ M A,KERSTEN S.Regulation of lipid droplet-associated proteins by peroxisome proliferator-activated receptors[J]. Biochim Biophys Acta Mol Cell Biol Lipids,2017,1862(10):1212-1220.
[23] SOENEN S,MARIMAN E C M,VOGELS N,et al.Relationship between perilipin gene polymorphisms and body weight and body composition during weight loss and weight maintenance[J].Physiol Behav,2009,96(4-5):723-728.
[24] TANSEY J T,SZTALRYD C,GRUIA-GRAY J,et al.Perilipin ablation results in a lean mouse with aberrant adipocyte lipolysis,enhanced leptin production,and resistance to diet-induced obesity[J].Proc Natl Acad Sci U S A,2001,98(11):6494-6499.
[25] KRAJEWSKA M,BANARES S,ZHANG E E,et al.Development of diabesity in mice with neuronal deletion of Shp2 tyrosine phosphatase[J]. Am J Pathol,2008,172(5):1312-1324.
[26] BANNO R,ZIMMER D,DE JONGHE B C,et al.PTP1B and SHP2 in POMC neurons reciprocally regulate energy balance in mice[J].J Clin Invest,2010,120(3):720-734.
[27] HE Z,ZHANG S S,MENG Q Y,et al.Shp2 controls female body weight and energy balance by integrating leptin and estrogen signals[J].Mol Cell Biol,2012,32(10):1867-1878.
[28] NAGATA N,MATSUO K,BETTAIEB A,et al.Hepatic Src homology phosphatase 2 regulates energy balance in mice[J]. Endocrinology, 2012,153(7):3158-3169.
[29] WOELLER C F,O'LOUGHLIN C W,POLLOCK S J,et al.Thy1(CD90) controls adipogenesis by regulating activity of the Src family kinase, Fyn[J].FASEB J,2015,29(3):920-931.
[30] WOELLER C F,FLORES E,POLLOCK S J,et al.THY1(CD90) expression is reduced by the environmental chemical tetrabromobisphenol-A to promote adipogenesis through induction of microRNA-103[J].Toxicol Sci,2017,157(2):305-319.
[31] 李江璇,肖丽玲,饶从强,等.人与大鼠脂肪干细胞提取方法及生物学特性的比较研究[J].暨南大学学报:自然科学与医学版,2015,36(4):324-329.
LI J X,XIAO L L,RAO C Q,et al.Comparison of isolation and biological characteristics of adipose-derived stem cells from human and rat[J].Journal of Jinan University:Natural Science & Medicine Edition,2015,36(4):324-329.(in Chinese)
[32] 王娜.人脂肪源性干细胞体外培养和鉴定研究[J].湖北科技学院学报:医学版,2016,30(6):468-470.
WANG N.Culture and identification of human adipose tissue derived stem cells in vitro[J].Journal of Hubei University of Science and Technology:Medical Sciences,2016,30(6):468-470.(in Chinese)
[33] 甘露,罗清礼,唐飞,等.TAO患者眼眶CD90⁺和CD90^-成纤维细胞亚群脂成纤维细胞转化的研究[J].四川大学学报:医学版,2016,47(3):356-359.
GAN L,LUO Q L,TANG F,et al.The study of lipofibroblasts differentiation of CD90⁺ and CD90^- orbital fibroblasts subsets in patients with TAO[J].Journal of Sichuan University:Medical Science Edition,2016,47(3):356-359.(in Chinese)
[34] LEE M W,CHOI J,YANG M S,et al.Mesenchymal stem cells from cryopreserved human umbilical cord blood[J].Biochem Biophys Res Commun,2004,320(1):273-278.
[35] YERLY S,GÜNTHARD H F,FAGARD C,et al.Proviral HIV-DNA predicts viral rebound and viral setpoint after structured treatment interruptions[J].AIDS,2004,18(14):1951-1953.
[36] WOOD I S,WANG B H,JENKINS J R,et al.The pro-inflammatory cytokine IL-18 is expressed in human adipose tissue and strongly upregulated by TNFα in human adipocytes[J].Biochem Biophys Res Commun,2005,337(2):422-429.
[37] SKURK T, KOLB H, MÜLLER-SCHOLZE S, et al.The proatherogenic cytokine interleukin-18 is secreted by human adipocytes[J]. Eur J Endocrinol,2005,152(6):863-868.
[38] KROGH-MADSEN R,PLOMGAARD P,MØLLER K,et al.Influence of TNF-alpha and IL-6 infusions on insulin sensitivity and expression of IL-18 in humans[J].Am J Physiol Endocrinol Metab,2006,291(1):E108-E114.
[39] BRUUN J M,STALLKNECHT B,HELGE J W,et al.Interleukin-18 in plasma and adipose tissue:effects of obesity,insulin resistance,and weight loss[J].Eur J Endocrinol,2007,157(4):465-471.
[40] ZILVERSCHOON G R C,TACK C J,JOOSTEN L A B,et al.Interleukin-18 resistance in patients with obesity and type 2 diabetes mellitus[J].Int J Obes (Lond),2008,32(9):1407-1414.
[41] GREGERSEN S, SAMOCHA-BONET D, HEILBRONN L K, et al.Inflammatory and oxidative stress responses to high-carbohydrate and high-fat meals in healthy humans[J].J Nutr Metab,2012,2012:238056.
[42] ESPOSITO K,NAPPO F,GIUGLIANO F,et al.Meal modulation of circulating interleukin 18 and adiponectin concentrations in healthy subjects and in patients with type 2 diabetes mellitus[J].Am J Clin Nutr,2003,78(6):1135-1140.
[43] KIM C W,MOON Y A,PARK S W,et al.Induced polymerization of mammalian acetyl-CoA carboxylase by MIG12 provides a tertiary level of regulation of fatty acid synthesis[J].Proc Natl Acad Sci U S A,2010,107(21):9626-9631.
[44] ZHU Q H,ANDERSON G W,MUCHA G T,et al.The Spot 14 protein is required for de novo lipid synthesis in the lactating mammary gland[J].Endocrinology,2005,146(8):3343-3350.
[45] RESNYK C W,CARRÉ W,WANG X F,et al.Transcriptional analysis of abdominal fat in genetically fat and lean chickens reveals adipokines,lipogenic genes and a link between hemostasis and leanness[J].BMC Genomics,2013,14:557.
[46] YIN L Y,ZHANG Y Q,CHARRON T,et al.Thyroid hormone,glucagon,and medium-chain fatty acids regulate transcription initiated from promoter 1 and promoter 2 of the acetyl-CoA carboxylase-α gene in chick embryo hepatocytes[J].Biochim Biophys Acta Gene Struct Exp,2000,1517(1):91-99.
[47] YIN L Y,ZHANG Y Q,HILLGARTNER F B.Sterol regulatory element-binding protein-1 interacts with the nuclear thyroid hormone receptor to enhance acetyl-CoA carboxylase-α transcription in hepatocytes[J].J Biol Chem,2002,277(22):19554-19565.
[48] AIPOALANI D L,O'CALLAGHAN B L,MASHEK D G,et al.Overlapping roles of the glucose-responsive genes,S14 and S14R,in hepatic lipogenesis[J].Endocrinology,2010,151(5):2071-2077.
[49] 李星艳,王世银,许瑞霞,等.阿勒泰羊CFD基因的克隆及其在不同营养状态阿勒泰羊尾脂中的表达分析[J].新疆农业科学,2016,53(6):1136-1144.
LI X Y,WANG S Y,XU R X,et al.Cloning of CFD gene in Altay sheep and analysis of expression levels in tail fat of Altay sheep in different nutrition states[J].Xinjiang Agricultural Sciences,2016,53(6):1136-1144.(in Chinese)
[50] ZEZULAK K M,GREEN H.Specificity of gene expression in adipocytes[J].Mol Cell Biol,1985,5(2):419-421.
[51] LEIHERER A,STOEMMER K,MUENDLEIN A,et al.Quercetin impacts expression of metabolism-and obesity-associated genes in SGBS adipocytes[J].Nutrients,2016,8(5):E282.
[52] SONG N J,KIM S,JANG B H,et al.Small molecule-induced complement Factor D (Adipsin) promotes lipid accumulation and adipocyte differentiation[J].PLoS One,2016,11(9):e0162228.
[53] GESTA S,TSENG Y H,KAHN C R.Developmental origin of fat:tracking obesity to its source[J].Cell,2007,131(2):242-256.
[54] YAMAMOTO Y,GESTA S,LEE K Y,et al.Adipose depots possess unique developmental gene signatures[J].Obesity (Silver Spring),2010,18(5):872-878.
[55] KARASTERGIOU K,FRIED S K,XIE J,et al.Distinct developmental signatures of human abdominal and gluteal subcutaneous adipose tissue depots[J].J Clin Endocrinol Metab,2013,98(1):362-371.
[56] LIU Z H,JI Z B,WANG G Z,et al.Genome-wide analysis reveals signatures of selection for important traits in domestic sheep from different ecoregions[J].BMC Genomics,2016,17:863.