鸭肠炎病毒感染鸭肝脏组织miRNA表达谱差异分析

Differential Analysis of miRNA Expression Profiles in Liver of Duck Infected with Duck Enteritis Virus

为探究鸭感染鸭肠炎病毒(Duck enteritis virus,DEV)后肝脏miRNA表达谱的差异,试验以DEV-GZ株经腿部肌肉接种30日龄麻鸭,于感染后66、90和114 h采集鸭肝脏组织样本,提取组织总RNA,经质检合格后,采用高通量测序技术对对照组和试验组样品进行miRNA测序,筛选出DEV感染鸭肝脏组织的差异表达miRNA,对其进行生物信息学GO功能分类和KEGG信号通路分析,并随机选取部分差异表达miRNA进行实时荧光定量PCR验证。结果显示,鸭感染DEV后66、90和114 h,肝脏组织差异表达miRNAs数量分别为227、225和231个。GO功能注释显示,感染鸭肝脏差异表达miRNA在生物过程分类中主要为细胞过程、单有机体过程和代谢过程类别;在细胞成分分类中主要是细胞、细胞部分和细胞器类别;在分子功能分类中主要是绑定分子功能和催化活性功能类别。KEGG通路富集显示,差异表达miRNA主要涉及PI3K-Akt、JAK-STAT、磷脂酰肌醇信号通路系统、ECM-受体相互作用、MAPK、Wnt、Toll样受体、IL-17、脂质代谢、钙离子信号通路和cAMP等信号通路,其中感染66 h差异表达miRNA主要在生物系统及神经系统中发挥作用;感染90 h主要在内分泌系统及消化系统中发挥作用;感染114 h主要在全身生物、免疫和消化系统等中发挥作用。选取10个差异表达miRNAs进行实时荧光定量PCR验证,结果与高通量测序结果一致。表明DEV感染对鸭肝脏组织miRNA表达具有显著影响,为从宿主miRNA角度揭示DEV致病机制提供了参考依据。

In order to explore the differential expression of miRNA in liver of duck infected by Duck enteritis virus(DEV),in this experiment,30-day-old hemp ducks were inoculated with the DEV-GZ strain through the leg muscles.Duck liver tissue samples were collected at 66,90 and 114 h after infection,and total RNA was extracted from the tissue.After passing the quality inspection,high-throughput sequencing technology performed miRNA sequencing on the samples of control and test groups,and screened out the differentially expressed miRNA in liver of DEV-infected duck,performed bioinformatics GO functional classification and KEGG signal pathway analysis,and randomly selected some differentially expressed miRNA for Real-time quantitative PCR verification.The results showed that when DEV infected ducks at 66,90 and 114 h,the number of differentially expressed miRNAs in liver were 227,225 and 231,respectively.GO function annotations showed that the differentially expressed miRNA in liver of infected ducks were mainly classified into cellular processes,single-organism processes and metabolic processes in the classification of biological processes.In the classification of cell components,they were mainly classified into cells,cell parts and organelles.In molecular functions,the classification mainly included binding molecular function and catalytic activity function category.KEGG pathway enrichment showed that differentially expressed miRNA mainly involved PI3K-Akt,JAK-STAT,phosphatidylinositol signaling pathway system,ECM-receptor interaction,MAPK,Wnt,Toll-like receptors,IL-17,lipid metabolism,calcium signal pathway,cAMP and other signaling pathways.Among them,differentially expressed miRNA mainly played a role in the biological system and nervous system after 66 h of infection,mainly played a role in endocrine system and digestive system after 90 h of infection,and mainly played a role in the systemic biology,immune and digestive system after 114 h of infection.10 differentially expressed miRNAs were selected for Real-time quantitative PCR verification,and the results were consistent with the results of high-throughput sequencing.It indicated that DEV infection had a significant impact on the expression of miRNA in liver of duck,and provided a reference for revealing the pathogenic mechanism of DEV from the perspective of host miRNA.