大型迪庆藏猪不同生长阶段背脂与腹脂脂质代谢差异基因及调控网络分析

doi:10.11843/j.issn.0366-6964.2023.02.010

畜牧兽医学报 ›› 2023, Vol. 54 ›› Issue (2): 520-533.doi: 10.11843/j.issn.0366-6964.2023.02.010

大型迪庆藏猪不同生长阶段背脂与腹脂脂质代谢差异基因及调控网络分析

王琳¹, 马黎², 张博³, 邓俊⁴, 张浩³, 欧阳晓芳², 严达伟^1*, 董新星^1*

1. 云南农业大学动物科学技术学院, 昆明 650201;
2. 云南农业职业技术学院, 昆明 650212;
3. 中国农业大学动物科学技术学院, 北京 100193;
4. 云南省畜牧总站, 昆明 650224

收稿日期:2022-07-12 出版日期:2023-02-23 发布日期:2023-02-21
通讯作者: 严达伟,主要从事动物遗传育种研究,E-mail:1302648630@qq.com;董新星,主要从事动物遗传育种研究,E-mail:86127447@qq.com
作者简介:王琳(1995-),女,河南温县人,硕士生,主要从事动物遗传育种与繁殖研究,E-mail:506874621@qq.com
基金资助:
云南省乡村振兴科技专项项目（202104BI090021）；云南省重大科技专项项目（202202AE090005）；云南省农业联合面上项目（2021BD070001-10）

Key Genes and Regulatory Network Analysis of Lipid Metabolism Differences between Back Fat and Abdominal Fat of Large Diqing Tibetan Pigs at Different Growth Stages

WANG Lin¹, MA Li², ZHANG Bo³, DENG Jun⁴, ZHANG Hao³, OUYANG Xiaofang², YAN Dawei^1*, DONG Xinxing^1*

1. College of Animal Science and Technology, Yunnan Agricultural University, Kunming 650201, China;
2. Yunnan Vocational and Technical College of Agriculture, Kunming 650212, China;
3. College of Animal Science and Technology, China Agricultural University, Beijing 100193, China;
4. Yunnan Province Animal Husbandry Station, Kunming 650224, China

Received:2022-07-12 Online:2023-02-23 Published:2023-02-21

摘要/Abstract

摘要： 旨在筛选大型迪庆藏猪不同生长阶段、不同部位脂质代谢差异的关键基因。本研究选择胎次相同、出生日期相近、体重10 kg左右的大型迪庆藏猪36头，随机分为3组，相同条件育肥，分别在平均体重达40、80和120 kg时屠宰，测定胴体性能，每组采集3头猪的背脂和腹脂进行高通量转录组测序，测序数据经拼接、比对，筛选与脂质代谢相关的显著差异基因并进行GO、KEGG分析、基因互作网络分析。结果表明，40、80和120 kg大型迪庆藏猪腹脂vs.背脂分别筛到486、765和339个差异表达显著基因，随机挑选的EGR2、SOD3等5个差异表达显著基因的qPCR结果与转录组测序结果一致，差异表达显著基因主要富集在肌肉收缩、细胞黏附、间充质细胞增殖正调节等GO条目，心肌收缩、PI3K-Akt信号通路、Hippo信号通路等KEGG通路；40 kg组EGR2、RARRES2、TMOD4和SFRP2基因位于网络核心，EGR2、RARRES2、SFRP2在腹脂上调，TMOD4下调；80 kg组THBS1、PPARA、NRIP1和LPL基因位于网络核心，4个核心基因均在腹脂上调；120 kg组HTRA1、TSHR、LRRK2、STC2、SHOX2和SOD3基因位于网络核心，LRRK2、TSHR在腹脂上调，SHOX2、SOD3、STC2、HTRA1下调。结果提示，EGR2、THBS1、TSHR等14个基因作为核心基因精细调控大型迪庆藏猪不同生长阶段背脂与腹脂脂质代谢，10～40 kg，EGR2等4个基因位于核心，促进脂肪细胞分化、增殖的基因在腹脂上调，脂肪合成的开关基因下调；40～80 kg，THBS1等4个基因位于核心，促进甘油三酯合成、胆固醇形成、脂质积累的基因在腹脂上调；80～120 kg，LRRK2等6个基因位于核心，促进甘油三酯积累、脂肪酸氧化的基因在腹脂上调，抑制脂肪分解、脂滴形成的基因下调。本试验结果可为解析地方猪不同部位脂质差异性沉积的调控机制提供基础数据，为迪庆藏猪的靶向选育提供参考。

关键词: 迪庆藏猪, 脂肪差异沉积, 功能基因, 基因互作网络

Abstract: The aim of this study was to screen the key genes of lipid metabolism differences in back fat and abdominal fat of large Diqing Tibetan pigs (TPs) at different growth stages by transcriptome sequencing. In this study, 36 TPs with the same parity, birth date and weight of about 10 kg were randomly divided into 3 groups for fattening experiment under the same conditions. The pigs were slaughtered at the weight of 40, 80 and 120 kg, respectively, and carcass perfor-mance was measured. Back fat (BF) and abdominal fat (AF) of 3 pigs in each group were collected for high-throughput transcriptome sequencing. The sequencing data were spliced and compared to screen for genes with significant differences related to lipid metabolism, and then GO, KEGG analysis and gene interaction network analysis were performed. The results showed that 486, 765 and 339 differentially expressed genes (DEGs) were screened from AF vs. BF of large Diqing Tibetan pigs weighing 40, 80 and 120 kg, respectively. The qPCR results of randomly selected 5 significantly DEGs such as EGR2 and SOD3 were consistent with the transcriptome sequencing results. These DEGs were mainly enriched in GO items such as muscle contraction, cell adhesion and positive regulation of mesenchymal cell proliferation, and KEGG pathways such as cardiac muscle contraction, PI3K-Akt signaling pathway, and Hippo signaling pathway. In 40 kg group, EGR2, RARRES2, TMOD4 and SFRP2 genes were located in the network core, EGR2, RARRES2 and SFRP2 were up-regulated in abdominal fat, and TMOD4 was down-regulated. In 80 kg group,THBS1, PPARA, NRIP1 and LPL genes were located in the core of the network, and all 4 core genes were up-regulated in AF. In 120 kg group, HTRA1, TSHR, LRRK2, STC2, SHOX2 and SOD3 genes were located in the network core, LRRK2 and TSHR were up-regulated in AF, while SHOX2, SOD3, STC2 and HTRA1 were down-regulated. The results suggested that 14 core genes including EGR2, THBS1 and TSHR finely regulated the lipid metabolism of BF and AF at different growth stages of TPs. 10-40 kg, 4 genes such as EGR2 were located in the network core, the genes promoting adipocyte differentiation and proliferation were up-regulated in AF, and the switch genes for fat synthesis were down-regulated. 40-80 kg, THBS1 and other 3 genes were located in the network core, and the genes promoting triglyceride synthesis, cholesterol formation and lipid accumulation were up-regulated in AF. 80-120 kg, 6 genes such as LRRK2 were located in the network core, the genes promoting triglyceride accumulation and fatty acid oxidation were up-regulated in AF, and the genes inhibiting fat decomposition and fat droplet formation were down-regulated. The results can provide basic data for the analysis of the regulation mechanism of differential fat deposition in different parts of local pigs, and provide reference for the targeted breeding of TPs.

Key words: Diqing Tibetan pigs, differential fat deposition, functional genes, gene interaction networks

中图分类号:

S828.2

王琳, 马黎, 张博, 邓俊, 张浩, 欧阳晓芳, 严达伟, 董新星. 大型迪庆藏猪不同生长阶段背脂与腹脂脂质代谢差异基因及调控网络分析[J]. 畜牧兽医学报, 2023, 54(2): 520-533.

WANG Lin, MA Li, ZHANG Bo, DENG Jun, ZHANG Hao, OUYANG Xiaofang, YAN Dawei, DONG Xinxing. Key Genes and Regulatory Network Analysis of Lipid Metabolism Differences between Back Fat and Abdominal Fat of Large Diqing Tibetan Pigs at Different Growth Stages[J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(2): 520-533.

参考文献 59

[1]	POULOS S P, HAUSMAN D B, HAUSMAN G J.The development and endocrine functions of adipose tissue[J].Mol Cell Endocrinol, 2010, 323(1):20-34.
[2]	SCHERER P E.Adipose tissue:from lipid storage compartment to endocrine organ[J].Diabetes, 2006, 55(6):1537-1545.
[3]	宋倩倩.肠道微生物与宿主基因互作对肉鸭腹脂沉积的遗传机制研究[D].扬州:扬州大学, 2021.SONG Q Q.Metagenomics and transcriptomics reveal the genetic mechanism of abdominal fat deposition in duck[D]. Yangzhou:Yangzhou University, 2021.(in Chinese)
[4]	HAUSMAN G J, BERGEN W G, ETHERTON T D, et al.The history of adipocyte and adipose tissue research in meat animals[J].J Anim Sci, 2018, 96(2):473-486.
[5]	有文静.GADD45α对动物脂肪细胞分化和代谢的影响及调控机制研究[D].杭州:浙江大学, 2020.YOU W J.Effects and regulatory mechanisms of GADD45α on differentiation and metabolism of animal adipocyte[D]. Hangzhou:Zhejiang University, 2020.(in Chinese)
[6]	谢光杰, 王永, 许晴, 等.简州大耳羊肌内脂肪细胞成脂分化差异表达基因的筛选与鉴定[J].畜牧兽医学报, 2020, 51(7):1525-1536.XIE G J, WANG Y, XU Q, et al.Selection and validation of the differentially expressed genes during the adipogenic differentiation of Jianzhou Da'er goat intramuscular adipocytes[J].Acta Veterinaria et Zootechnica Sinica, 2020, 51(7):1525-1536.(in Chinese)
[7]	YANG H, XU X L, MA H M, et al.Integrative analysis of transcriptomics and proteomics of skeletal muscles of the Chinese indigenous Shaziling pig compared with the Yorkshire breed[J].BMC Genet, 2016, 17(1):80.
[8]	JIN L, TANG Q Z, HU S L, et al.A pig BodyMap transcriptome reveals diverse tissue physiologies and evolutionary dynamics of transcription[J].Nat Commun, 2021, 12(1):3715.
[9]	CHEN C Y, AI H S, REN J, et al.A global view of porcine transcriptome in three tissues from a full-sib pair with extreme phenotypes in growth and fat deposition by paired-end RNA sequencing[J].BMC Genomics, 2011, 12:448.
[10]	WANG T, JIANG A A, GUO Y Q, et al.Deep sequencing of the transcriptome reveals inflammatory features of porcine visceral adipose tissue[J].Int J Biol Sci, 2013, 9(6):550-556.
[11]	MERRETT J E, BO T, PSALTIS P J, et al.Identification of DNA response elements regulating expression of CCAAT/enhancer-binding protein (C/EBP) β and δ and MAP kinase-interacting kinases during early adipogenesiss[J].Adipocyte, 2020, 9(1):427-442.
[12]	SHIMOMURA I, BASHMAKOV Y, IKEMOTO S, et al.Insulin selectively increases SREBP-1c mRNA in the livers of rats with streptozotocin-induced diabetes[J].Proc Natl Acad Sci U S A, 1999, 96(24):13656-13661.
[13]	DELGHANDI M P, JOHANNESSEN M, MOENS U.The cAMP signalling pathway activates CREB through PKA, p38 and MSK1 in NIH 3T3 cells[J].Cell Signal, 2005, 17(11):1343-1351.
[14]	HOMAN E P, BRANDÃO B B, SOFTIC S, et al.Differential roles of FOXO transcription factors on insulin action in brown and white adipose tissue[J].J Clin Invest, 2021, 131(19):e143328.
[15]	RAZA S H A, PANT S D, WANI A K, et al.Krüppel-like factors family regulation of adipogenic markers genes in bovine cattle adipogenesis[J].Mol Cell Probes, 2022, 65:101850.
[16]	LI D, ZHANG F, ZHANG X, et al.Distinct functions of PPARγ isoforms in regulating adipocyte plasticity[J].Biochem Biophys Res Commun, 2016, 481(1-2):132-138.
[17]	李优磊.PPAR信号通路在调控猪皮下脂肪与肌内脂肪差异沉积中的作用及机制研究[D].杨凌:西北农林科技大学, 2018.LI Y L.The differential function and mechanism of PPAR signaling in regulation of porcine subcutaneous and intramuscular fat depositon[D].Yangling:Northwest A&F University, 2018.(in Chinese)
[18]	严达伟, 赵桂英, 苟潇, 等.迪庆藏猪肉质特性的研究[J].云南农业大学学报, 2007, 22(1):86-91.YAN D W, ZHAO G Y, GOU X, et al.Study on characteristics of Diqing Tibetan Pig's meat quality[J].Journal of Yunnan Agricultural University, 2007, 22(1):86-91.(in Chinese)
[19]	聂靖茹, 张博, 马黎, 等.大型迪庆藏猪不同生长阶段肌肉生长差异基因及其调控通路分析[J].中国农业大学学报, 2022, 27(6):132-144.NIE J R, ZHANG B, MA L, et al.Screening of differential genes for muscle growth and regulatory pathways analysis in large Diqing Tibetan pig at different growth stages[J].Journal of China Agricultural University, 2022, 27(6):132-144.(in Chinese)
[20]	聂靖茹, 马黎, 鲁绍雄, 等.迪庆藏猪与野猪×迪庆藏猪生长、胴体及肉质性能比较[J].云南农业大学学报:自然科学, 2021, 36(5):805-810.NIE J R, MA L, LU S X, et al.Comparison of growth, carcass and meat quality traits between Diqing Tibetan pig and wild boar×Diqing Tibetan pig[J].Journal of Yunnan Agricultural University (Natural Science), 2021, 36(5):805-810.(in Chinese)
[21]	易胜男, 孔小艳, 钱锦花, 等.藏猪EPAS1基因低氧适应相关位点与表达量研究[J].家畜生态学报, 2020, 41(6):19-24.YI S N, KONG X Y, QIAN J H, et al.Related loci and expression level of EPAS1 gene on hypoxia adaptation in Tibetan pigs[J].Journal of Domestic Animal Ecology, 2020, 41(6):19-24.(in Chinese)
[22]	KURI-HARCUCH W, VELEZ-DELVALLE C, VAZQUEZ-SANDOVAL A, et al.A cellular perspective of adipogenesis transcriptional regulation[J].J Cell Physiol, 2019, 234(2):1111-1129.
[23]	PENG Y D, XIANG H, CHEN C, et al.MiR-224 impairs adipocyte early differentiation and regulates fatty acid metabolism[J].Int J Biochem Cell Biol, 2013, 45(8):1585-1593.
[24]	CHEN Z, TORRENS J I, ANAND A, et al.Krox20 stimulates adipogenesis via C/EBPβ-dependent and -independent mechanisms[J]. Cell Metab, 2005, 1(2):93-106.
[25]	JIANG Y Q, LIU P, JIAO W L, et al.Gaxsuppresses chemerin/CMKLR1-induced preadipocyte biofunctions through the inhibition of Akt/mTOR and ERK signaling pathways[J].J Cell Physiol, 2018, 233(1):572-586.
[26]	CATALÁN V, GÓMEZ-AMBROSI J, RODRÍGUEZ A, et al.Increased levels of chemerin and its receptor, chemokine-like receptor-1, in obesity are related to inflammation:tumor necrosis factor-α stimulates mRNA levels of chemerin in visceral adipocytes from obese patients[J].Surg Obes Relat Dis, 2013, 9(2):306-314.
[27]	HUANG C L, XIAO L L, XU M, et al.Chemerin deficiency regulates adipogenesis is depot different through TIMP1[J].Genes Dis, 2021, 8(5):698-708.
[28]	EHRLUND A, MEJHERT N, LORENTE-CEBRIÁN S, et al.Characterization of the Wnt inhibitors secreted frizzled-related proteins (SFRPs) in human adipose tissue[J].J Clin Endocrinol Metab, 2013, 98(3):E503-E508.
[29]	BOLAMPERTI S, SIGNO M, SPINELLO A, et al.GH prevents adipogenic differentiation of mesenchymal stromal stem cells derived from human trabecular bone via canonical Wnt signaling[J].Bone, 2018, 112:136-144.
[30]	CROWLEY R K, O'REILLY M W, BUJALSKA I J, et al.SFRP2 is associated with increased adiposity and VEGF expression[J].PLoS One, 2016, 11(9):e0163777.
[31]	ZHAO X, HUANG Z, LIU X H, et al.The switch role of the Tmod4 in the regulation of balanced development between myogenesis and adipogenesis[J].Gene, 2013, 532(2):263-271.
[32]	MATSUO Y, TANAKA M, YAMAKAGE H, et al.Thrombospondin 1 as a novel biological marker of obesity and metabolic syndrome[J].Metabolism, 2015, 64(11):1490-1499.
[33]	RAMIS J M, HAL N L W F V, KRAMER E, et al.Carboxypeptidase E and thrombospondin-1 are differently expressed in subcutaneous and visceral fat of obese subjects[J].Cell Mol Life Sci, 2002, 59(11):1960-1971.
[34]	LI Y Z, TONG X P, RUMALA C, et al.Thrombospondin1 deficiency reduces obesity-associated inflammation and improves insulin sensitivity in a diet-induced obese mouse model[J].PLoS One, 2011, 6(10):e26656.
[35]	TIAN X, RU Q, XIONG Q, et al.Catalpol attenuates hepatic steatosis by regulating lipid metabolism via AMP-activated protein kinase activation[J].BioMed Res Int, 2020, 2020:6708061.
[36]	SUN H J, ZHU X X, CAI W W, et al.Hypaphorine attenuates lipopolysaccharide-induced endothelial inflammation via regulation of TLR4 and PPAR-γ dependent on PI3K/Akt/mTOR signal pathway[J].Int J Mol Sci, 2017, 18(4):844.
[37]	AUGEREAU P, BADIA E, CARASCOSSA S, et al.The nuclear receptor transcriptional coregulator RIP140[J].Nucl Recept Signal, 2006, 4:e024.
[38]	DE MARINIS Y, SUN J M, BOMPADA P, et al.Regulation of nuclear receptor interacting protein 1 (NRIP1) gene expression in response to weight loss and exercise in humans[J].Obesity (Silver Spring), 2017, 25(8):1400-1409.
[39]	CAVAILLōS V, DAUVOIS S, DANIELIAN P S, et al.Interaction of proteins with transcriptionally active estrogen receptors[J]. Proc Natl Acad Sci U S A, 1994, 91(21):10009-10013.
[40]	NAUTIYAL J.Transcriptional coregulator RIP140:an essential regulator of physiology[J].J Mol Endocrinol, 2017, 58(3):R147-R158.
[41]	ZSCHIEDRICH I, HARDELAND U, KRONES-HERZIG A, et al.Coactivator function of RIP140 for NFκB/RelA-dependent cytokine gene expression[J].Blood, 2008, 112(2):264-276.
[42]	HU Y, ZHU Y, GERBER S D, et al.Deletion of Nrip1 delays skin aging by reducing adipose-derived mesenchymal stem cells (ADMSCs) senescence, and maintaining ADMSCs quiescence[J].Geroscience, 2021, 43(4):1815-1833.
[43]	LEONARDSSON G, STEEL J H, CHRISTIAN M, et al.Nuclear receptor corepressor RIP140 regulates fat accumulation[J].Proc Natl Acad Sci U S A, 2004, 101(22):8437-8442.
[44]	NIMONKAR A V, WELDON S, GODBOUT K, et al.A lipoprotein lipase-GPI-anchored high-density lipoprotein-binding protein 1 fusion lowers triglycerides in mice:implications for managing familial chylomicronemia syndrome[J].J Biol Chem, 2020, 295(10):2900-2912.
[45]	OLIVECRONA G.Role of lipoprotein lipase in lipid metabolism[J].Curr Opin Lipidol, 2016, 27(3):233-241.
[46]	HE P P, JIANG T, OUYANG X P, et al.Lipoprotein lipase:biosynthesis, regulatory factors, and its role in atherosclerosis and other diseases[J].Clin Chim Acta, 2018, 480:126-137.
[47]	SEOL W, NAM D, SON I.Rab GTPases as physiological substrates of LRRK2 kinase[J].Exp Neurobiol, 2019, 28(2):134-145.
[48]	LIN C W, PENG Y J, LIN Y Y, et al.LRRK2 regulates CPT1A to promote β-oxidation in HepG2 cells[J].Molecules, 2020, 25(18):4122.
[49]	KHAN S, CHAN Y T, REVELO X S, et al.The immune landscape of visceral adipose tissue during obesity and aging[J].Front Endocrinol (Lausanne), 2020, 11:267.
[50]	YAN F, WANG Q, LU M, et al.Thyrotropin increases hepatic triglyceride content through upregulation of SREBP-1c activity[J].J Hepatol, 2014, 61(6):1358-1364.
[51]	LU S M, GUAN Q B, LIU Y T, et al.Role of extrathyroidal TSHR expression in adipocyte differentiation and its association with obesity[J].Lipids Health Dis, 2012, 11(1):17.
[52]	ZHANG J M, WU H X, MA S Z, et al.TSH promotes adiposity by inhibiting the browning of white fat[J].Adipocyte, 2020, 9(1):264-278.
[53]	LEE K Y, YAMAMOTO Y, BOUCHER J, et al.Shox2 is a molecular determinant of depot-specific adipocyte function[J].Proc Natl Acad Sci U S A, 2013, 110(28):11409-11414.
[54]	GAO D, HU S J, ZHENG X W, et al.SOD3 is secreted by adipocytes and mitigates high-fat diet-induced obesity, inflammation, and insulin resistance[J].Antioxid Redox Signal, 2020, 32(3):193-212.
[55]	CUI R, GAO M, QU S, et al.Overexpression of superoxide dismutase 3 gene blocks high-fat diet-induced obesity, fatty liver and insulin resistance[J].Gene Ther, 2014, 21(9):840-848.
[56]	MA B C, XU X Y, HE S, et al.STC2 modulates ERK1/2 signaling to suppress adipogenic differentiation of human bone marrow mesenchymal stem cells[J].Biochem Biophys Res Commun, 2020, 524(1):163-168.
[57]	ZHAO J J, JIAO Y, SONG Y P, et al.Stanniocalcin 2 ameliorates hepatosteatosis through activation of STAT3 signaling[J].Front Physiol, 2018, 9:873.
[58]	TIADEN A N, BREIDEN M, MIRSAIDI A, et al.Human serine protease HTRA1 positively regulates osteogenesis of human bone marrow-derived mesenchymal stem cells and mineralization of differentiating bone-forming cells through the modulation of extracellular matrix protein[J].Stem Cells, 2012, 30(10):2271-2282.
[59]	TIADEN A N, BAHRENBERG G, MIRSAIDI A, et al.Novel function of serine protease HTRA1 in inhibiting adipogenic differentiation of human mesenchymal stem cells via MAP kinase-mediated MMP upregulation[J].Stem Cells, 2016, 34(6):1601-1614.

大型迪庆藏猪不同生长阶段背脂与腹脂脂质代谢差异基因及调控网络分析

Key Genes and Regulatory Network Analysis of Lipid Metabolism Differences between Back Fat and Abdominal Fat of Large Diqing Tibetan Pigs at Different Growth Stages

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