基于转录组学分析性别、月龄对荷斯坦牛网膜脂肪组织炎症相关基因表达的影响  

Effects of Sex and Age on Inflammation-related Genes Expression in Omental Adipose Tissue of Holstein Cattle(Bos taurus)Based on Transcriptomics

作　　者：柴金宝 刘利[1] 王芳[1] 杨硕 曹培丽 赵晓川[1] 许珊珊[1] 孟详人[1] 卜也 岳萌萌 武瑞[2] 孙芳[1] CHAI Jin-Bao;LIU Li;WANG Fang;YANG Shuo;CAO Pei-Li;ZHAO Xiao-Chuan;XU Shan-Shan;MENG Xiang-Ren;BU Ye;YUE Meng-Meng;WU Rui;SUN Fang(Key Laboratory of Combining Farming and Animal Husbandry of Ministry of Agriculture,Institute of Animal Husbandry,Heilongjiang Academy of Agricultural Sciences,Harbin 150086,China;College of Animal Science and Veterinary Medicine,Heilongjiang Bayi Agricultural University,Daqing 163319,China)

机构地区：[1]黑龙江省农业科学院畜牧研究所/农业农村部种养结合重点实验室,哈尔滨150086 [2]黑龙江八一农垦大学动物科技学院,大庆163319

出　　处：《农业生物技术学报》2023年第5期901-913,共13页Journal of Agricultural Biotechnology

基　　金：国家肉牛牦牛产业技术体系建设(CARS37);国家自然科学基金青年基金(31702104);黑龙江省农业科技创新工程(2014ZD015);黑龙江省博士后资助经费(LBH-Z15198);黑龙江省现代农业产业技术协同创新推广体系建设(2021-No.1492)。

摘　　要：公牛(Bos taurus)去势和长期育肥能够促进内脏脂肪的沉积,过多的脂肪沉积易引发机体慢性炎症,然而有关牛网膜脂肪组织炎症的分子机理研究相对较少。本研究以牛网膜脂肪组织为研究对象,旨在探究性别和月龄对荷斯坦育肥牛网膜脂肪组织炎症相关基因表达的影响。选择16月龄荷斯坦公牛(BF16)、阉牛(SF16)及26月龄阉牛(SF26)各3头,测定生长和血液生化指标,屠宰后采集网膜脂肪组织,进行组织切片苏木精-伊红染色(hematoxylin-eosin staining,HE)染色,采用RNA-seq技术对3组牛网膜脂肪组织进行转录组学分析,筛选组间差异基因,并对差异基因进行GO功能注释和KEGG富集分析、qPCR验证、候选基因筛选及蛋白网络互作分析。结果显示,共得到1173个组间差异表达基因,富集在8个基因表达趋势模式中;功能分析显示差异基因富集多个与炎症相关的生物学过程和信号通路;随机选择6个差异表达的基因进行qPCR验证,表达趋势与转录组测序结果一致,表明测序结果可靠;筛选到与炎症相关的5个生物学过程条目和5个信号通路共50个非冗余基因,聚类分析发现,与BF16组相比,去势能够上调SF16组网膜脂肪组织中抗炎症和促脂肪细胞分化基因(如TNF-α诱导蛋白3(TNF alpha induced protein 3,TNFAIP3),骨形成蛋白2(bone morphogenetic protein 2,BMP2)等)的表达,下调促炎症基因(如白细胞介素1β(interleukin-1beta,IL1B))的表达;与BF16和SF16组相比,长期育肥能够上调SF26组网膜脂肪组织中衰老和慢性炎症相关的基因(如半乳糖凝集素3(galectin 3,LGALS3),C-C基序趋化因子受体7(C-C motif chemokine receptor 7,CCR7)等)的表达。蛋白网络互作分析显示IL1B为关键中心节点。本研究结果为优化利用荷斯坦阉牛生产大理石纹牛肉技术提供理论依据。Castration and long term fattening of bulls(Bos taurus)could promote visceral fat deposition,excessive fat deposition was easy to cause systemic chronic inflammation.However,there was very little known molecular mechanism of omental adipose tissue inflammation in cattle.The present study selected omental adipose tissue of cattle as research object,the aim of the present study was to evaluate the effects of sex and age on inflammation-related genes expression in omental adipose tissue of Holstein fattening cattle.Holstein bulls at 16 months of age(BF16 group,n=3)and Holstein steers at 16 and 26 months of age(SF16 and SF26 group,n=3)were selected.The growth performance and blood biochemical indexes were measured.After slaughtering,omental adipose tissue samples were collected for hematoxylin-eosin staining(HE)staining,transcriptome analysis was carried out for omental adipose tissue from three groups cattle by RNAseq.Differentially expressed genes(DEGs)between groups was screened,GO function annotation,KEGG enrichment analysis,qPCR validation,candidate gene screening and protein-protein interaction network of differentially expressed genes were conducted.The results showed that a total 1173 differentially expressed genes were identified,and enriched in 8 gene expression trend profiles.Functional analysis showed that the differentially genes were involved in multiple biological processes and signal pathways related to inflammation.Six differentially expressed genes were randomly selected for qPCR validation,and the results were consistent with transcriptome sequencing,which indicated that the sequencing results were reliable.Fifty non-redundant genes in 5 biological process items and 5 signaling pathways related to inflammation were screened.Cluster analysis showed that castration could upregulate the expression of anti-inflammatory and adipocyte differentiation genes(such as TNF alpha induced protein 3(TNFAIP3),bone morphogenetic protein 2(BMP2),etc)and downregulate the expression of proinflammatory genes(such as i

关 键 词：荷斯坦牛 转录组 性别 月龄 脂肪组织炎症 

分 类 号：S823.2[农业科学—畜牧学]