MDRV弱毒感染下番鸭肝脏源外泌体差异miRNA表达谱研究  

Differential expression profiles of miRNA in exosomes derived from hepatocytes of muscovy ducks immunized by live vaccine MDRV infection

作　　者：魏祥端 江珊 张蕾 王全溪[1] Wei Xiangduan;Jiang Shan;Zhang Lei;Wang Quanxi(College of Animal Science,Fujian Agriculture and Forestry University,Fuzhou350002)

机构地区：[1]福建农林大学动物科学学院,福州350002

出　　处：《福建畜牧兽医》2024年第4期13-18,共6页Fujian Journal of Animal Husbandry and Veterinary medicine

基　　金：福建省大学生创新训练项目(S202310389083)资助。

摘　　要：为探究番鸭呼肠孤病毒(Muscovy duck reovirus,MDRV)活疫苗对番鸭肝脏源外泌体中microRNA(miRNA)的影响,选择40羽1日龄雏番鸭,随机分为试验组和对照组,每组各20羽。试验组肌肉注射MDRV活疫苗1羽份,对照组肌肉注射等量的灭菌生理盐水。免疫48 h后,采集肝脏制备肝原代细胞培养2 d,取上清用试剂盒法提取外泌体送生物公司进行miRNA测序。接着进行miRNA注释,差异表达miRNA分析,靶基因预测,并对靶基因涉及的生物功能进行GO和KEGG富集分析。结果表明:MDRV活疫苗免疫组与对照组相比,共筛选到显著差异miRNA 39个,预测到369个靶基因,这些靶基因的GO功能注释主要集中在单有机体过程、刺激应答、多细胞有机体过程、定位、细胞器、膜、胞外区、绑定分子功能等;KEGG通路富集分析集中在肝脏中氨基糖和核苷酸糖的代谢、糖酵解/糖异生、溶酶体、磷脂酰肌醇信号传导系统、磷酸肌醇代谢、细胞凋亡、细胞自噬作用等。结果表明,MDRV活疫苗免疫雏番鸭会影响肝细胞外泌体的miRNA表达水平,这些miRNA参与肝脏代谢等多种功能。研究结果可为MDRV感染的科学防控提供理论基础。To explore the effect of live vaccine muscovy duck reovirus(MDRV)on microRNA(miRNA)in hepatocytes-derived exosomes of muscovy duck,forty 1-day-old muscovy ducklings were randomly divided into experimental group and control group,with 20 ducks in each group.The experimental group was intramuscularly injected with 1 dose of MDRV live vaccine,and the control group was injected with the same amount of sterilized normal saline.At 48 hours post immunization,the livers were collected to prepare primary liver cells and they were culture for 2 days,the exosomes from supernatant were extracted by exosome extraction kit,and were sent to a biological company for miRNA sequencing,annotation,differential expression analysis,target gene prediction,GO and KEGG enrichment analysis of the biological functions involved in the target genes were performed.The results showed that a total of 39 significantly different expression miRNAs were screened in the live vaccine MDRV immunization group compared with the control group,which targeted 369 mRNA.The GO functional annotations mainly focused on single-organism processes,stimulus responses,multicellular organism processes,localization,organelles,membranes,extracellular regions,and binding molecular functions.The result of KEGG pathway enrichment analysis mainly involved in the metabolism of amino sugars and nucleotide sugars,glycolysis/gluconeogenesis,lysosomes,phosphatidylinositol signaling system,inositol phosphate metabolism,apoptosis,and cell autophagocytosis in the liver.These results revealed that muscovy ducklings immunized with live vaccine MDRV regulated the expression level of miRNAs in hepatocyte exosomes,which were involved in liver metabolism and other functions.The results of this study will provide a theoretical basis for the scientific prevention and control of MDRV infection.

关 键 词：番鸭 番鸭呼肠孤病毒 肝细胞 外泌体 

分 类 号：S858.32[农业科学—临床兽医学]