生物技术通报 ›› 2021, Vol. 37 ›› Issue (5): 56-66.doi: 10.13560/j.cnki.biotech.bull.1985.2020-0919

• 研究报告 • 上一篇    下一篇

无菌猪和普通猪早期脂肪发育及脂肪组织基因转录表达的差异

邱小宇1(), 刘作华1,2, 齐仁立1,3()   

  1. 1.重庆市畜牧科学院,重庆 402460
    2.农业农村部养猪科学重点实验室,重庆 402460
    3.重庆市养猪科学重点实验室,重庆 402460
  • 收稿日期:2020-07-22 出版日期:2021-05-26 发布日期:2021-06-11
  • 作者简介:邱小宇,男,硕士研究生,研究方向:动物肉品质调控;E-mail: 1129363375@qq.com
  • 基金资助:
    国家重点研发计划项目(2018YFD0500404);重庆市财政资金专项项目(19513)

Differences in Early Fat Development and Gene Transcription Expression in the Adipose Tissues of Piglets with and Without Gut Microbiota

QIU Xiao-yu1(), LIU Zuo-hua1,2, QI Ren-li1,3()   

  1. 1. Chongqing Academy of Animal Sciences,Chongqing 402460
    2. Key Laboratory of Pig Industry Science,Ministry of Agriculture and Rural Affairs,Chongqing 402460
    3. Chongqing Key Laboratory of Pig Industry Science,Chongqing 402460
  • Received:2020-07-22 Published:2021-05-26 Online:2021-06-11

摘要:

通过比较无菌仔猪和普通仔猪在生长早期脂肪沉积的差异及脂肪组织基因转录表达谱的不同,评估肠道微生物定植对猪脂肪组织早期发育的直接影响。分别采集25日龄无菌仔猪(3头)和普通仔猪(3头)的颈部皮下脂肪,切片后经HE染色观察不同猪脂肪细胞的形态差异;Western Blot检测脂肪合成调控因子的蛋白表达差异;ELISA法测定不同猪脂肪组织分泌产生的脂肪细胞因子含量;转录组测序(RNA-seq)分析不同猪脂肪组织中的基因表达谱,鉴定关键差异基因及其相关信号网络。与普通仔猪相比,相同日龄的无菌仔猪体脂沉积量较少,皮下脂肪厚度和脂肪细胞尺寸均明显减小(P<0.001);脂肪组织中的脂肪酸结合蛋白4(fatty acid binding protein 4,FABP4)、过氧化物酶体增殖体激活受体γ(peroxisome proliferators activated recepor γ,PPARγ)、乙酰辅酶A羧化酶(acetyl CoA carboxylase,ACC)、脂肪酸合成酶(fatty acid synthase,FAS)的蛋白表达水平显著降低(P<0.05)。脂肪产生的功能性细胞因子脂联素(adiponectin,P=0.100)和瘦素(leptin,P=0.095)有着不同程度的减少。测序结果显示与普通猪相比,无菌猪的脂肪中有695个基因显著差异表达(P<0.05),包括338个基因表达上调,357个基因表达下调。qRT-PCR验证了转录组测序结果的准确性。KEGG通路富集分析结果显示差异基因主要与脂质合成代谢、分解代谢、脂质氧化、能量稳态相关。肠道微生物的定植显著影响了动物脂肪组织的发育、代谢和生理功能。

关键词: 肠道微生物, 脂肪, 基因, 转录组,

Abstract:

We compared the difference of fat deposition in the early growth stage and the different transcriptional profiles of adipose tissue in sterile piglets and normal piglets,and evaluated the direct effects of intestinal microbial colonization on the early development of pig adipose tissue. The cervical subcutaneous fat of 25-day sterile piglets(3 in GF piglet group)and normal pigs(3 in normal piglet group)was collected. HE staining was used to observe the morphological differences of fat cells,Western Blot to detect the protein expression differences of fat synthesis regulators,ELISA to detect the content of cytokines secreted in fat tissues,and RNA-seq to analyze the gene expression profile differences in fat tissues and identify the key differential genes and relevant signal networks. Compared with the normal piglets,the fat deposition in the same age of the sterile piglets was less,and the thickness of back fat and the sizes of fat cells significantly reduced(P<0.01). The expression of fatty acid binding protein 4(FABP4),peroxisome proliferators activated receptor γ(PPARγ),acetyl CoA carboxylase(ACC)and fatty acid synthase(FAS)in the fat decreased significantly(P<0.05). Adiponectin(P=0.100)and leptin(P=0.095)also decreased in varied degrees. The results of transcriptome sequencing showed that there were 695 genes in the fat of sterile piglets,including 338 genes up-regulated and 357 genes down-regulated(P<0.05,fold change>2). Accuracy of RNA-sequencing data was verified by the qRT-PCR results. KEGG analysis showed that the differentially expressed genes were mainly related to lipid synthesis metabolism,catabolism,lipid oxidation and energy homeostasis. The colonization and status of intestinal microorganisms significantly affect the development,metabolism and function of animal adipose tissue.

Key words: gut microorganism, fat, gene, transcriptome, pig