多浪羊与小尾寒羊皮下脂肪组织转录组分析

doi:10.11843/j.issn.0366-6964.2021.012.008

摘要/Abstract

摘要： 旨在探索多浪羊（D组）与小尾寒羊（X组）皮下脂肪组织中的特异性表达基因，并研究其潜在的作用，为理解绵羊脂肪组织发育规律以及对脂代谢相关疾病的预防和治疗研究提供依据。本研究选取脂肪沉积能力存在差异、健康无病、体况良好、种内个体体重相近（约50 kg）的雌性成年多浪羊和小尾寒羊为试验材料，分为D组（试验组）和X组（对照组），每组3个重复，采集位于背最长肌的皮下脂肪组织，应用RNA-Seq技术和生物信息学方法进行转录组测序并对结果进行分析。以|Fold change|≥2，P adjust≤0.05为标准筛选差异表达基因，通过对差异表达基因进行GO功能注释和KEGG通路富集分析，得到与脂肪沉积和脂代谢有关的差异基因。为了验证测序数据的可靠性，本研究随机选取了6个差异表达基因进行qRT-PCR验证。结果显示，在6个样本中共检测到38 672个已知的mRNAs，新的mRNAs为1 606个，在两组中共有839个差异表达基因，其中有320个差异基因在多浪羊组中上调表达，有519个差异基因在多浪羊组中下调表达。通过GO功能注释分析发现，差异表达基因主要参与脂质分解代谢过程、脂质生物合成过程、脂质分解代谢负调控过程、MAPK级联反应调控、对甘油三酯的反应等生物学过程。KEGG通路富集结果显示，差异表达基因显著富集到了PI3K-Akt、MAPK、胰岛素以及PPAR等信号通路中。qRT-PCR结果与测序结果一致，表明测序结果可靠。通过对多浪羊和小尾寒羊皮下脂肪组织进行转录组测序以及生物信息学分析，筛选到与脂肪沉积和脂代谢相关的差异表达基因，这些基因主要参与脂质生物合成、脂质代谢等过程，其中COL1A1、AKT2、SCD、LPL、PCK1与PPP2R5A可能在多浪羊与小尾寒羊的皮下脂肪组织的沉积与代谢中发挥重要作用。

关键词: mRNA, 皮下脂肪, 脂肪沉积, 脂质代谢, 绵羊

Abstract: The aim of this study was to explore the differentially expressed genes in the subcutaneous adipose tissue of Duolang sheep (group D) and Small Tail Han sheep (group X), study their potential effects, and to provide a basis for understanding the developmental law of sheep fat tissue and the prevention and treatment of lipid metabolism-related diseases. In the study, healthy, female adult Duolang sheep and Small Tail Han sheep with good body condition, similar in-species body weight (about 50 kg) and differences in fat deposition ability were selected as the test materials, and they were divided into group D (experimental group) and group X (control group), with 3 replicates in each group. The subcutaneous adipose tissue located in the longissimus dorsi muscle was collected. RNA-Seq technology and bioinformatics methods were used to perform the transcriptome sequencing and the results analysis. According to the criteria of|Fold change| ≥ 2 and P adjust ≤ 0.05, differentially expressed genes were screened. The differentially expressed mRNAs were analyzed by GO and KEGG enrichment to obtain the differential genes related to fat deposition and lipid metabolism. Meantime, the reliability of the sequencing data was verified by real-time fluorescent quantitative PCR for 6 randomly selected differentially expressed genes. The results showed that a total of 38 672 known mRNAs and 1 606 new mRNAs were detected in the 6 samples. There were 839 differentially expressed genes in the two groups, of which 320 were up-regulated and 519 were down-regulated in Duolang sheep. Through GO function annotation analysis, it was found that differentially expressed genes were mainly involved in lipid catabolism process, lipid biosynthesis process, lipid catabolism negative regulation process, MAPK cascade reaction regulation, and reaction to triglycerides. The enrichment results of KEGG pathways showed that PI3K-Akt, MAPK, insulin and PPAR signaling pathways were significantly enriched. The qRT-PCR results were consistent with the sequencing results, indicating that the sequencing results were reliable. Through transcriptome sequencing and bioinformatics analysis of subcutaneous fat tissue of Duolang sheep and Small Tail Han sheep, differentially expressed genes related to fat deposition and lipid metabolism were screened, these genes mainly involved in lipid biosynthesis, lipid metabolism and other processes. Among them, COL1A1, AKT2, SCD, LPL, PCK1 and PPP2R5A might play important roles in the deposition and metabolism of subcutaneous fat tissue in Duolang sheep and Small Tail Han sheep.

Key words: mRNA, subcutaneous fat, fat deposition, lipid metabolism, sheep

中图分类号:

S826.2

刘天义, 冯卉, Salsabeel Yousuf, 解领丽, 苗向阳. 多浪羊与小尾寒羊皮下脂肪组织转录组分析[J]. 畜牧兽医学报, 2021, 52(12): 3403-3412.

LIU Tianyi, FENG Hui, Salsabeel Yousuf, XIE Lingli, MIAO Xiangyang. Transcriptome Analysis of Subcutaneous Adipose Tissue of Duolang Sheep and Small Tail Han Sheep[J]. Acta Veterinaria et Zootechnica Sinica, 2021, 52(12): 3403-3412.

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