鸭瘟病毒感染鸭十二指肠黏膜炎症相关基因及调控通路的筛选

doi:10.16431/j.cnki.1671-7236.2020.10.032

摘要/Abstract

摘要： 试验旨在分析鸭瘟病毒（duck plague virus，DPV）感染樱桃谷鸭后十二指肠黏膜转录组水平的变化，筛选出DPV感染后参与炎症的相关基因和调控通路。DEV-GZ株经腿部肌肉接种35日龄鸭后，于接种后24、48和72 h采集鸭十二指肠黏膜组织样本，提取总RNA进行转录组测序，对数据进行质控与注释，筛选与炎症相关的差异表达基因及调控通路，并应用实时荧光定量PCR对部分差异表达基因进行验证。结果显示，对照组与试验组总计得到21.39 Gb的Clean Bases，测序样本有效序列占原始序列的99%以上，映射率高于83%。鸭接毒DPV后24 h十二指肠黏膜的差异表达基因有221个，参与炎症相关生物学过程的有3个，均为下调基因；接毒DPV后48 h差异表达基因有499个，参与炎症相关生物学过程的有17个，均为下调基因；接毒DPV后72 h差异表达基因有798个，参与炎症相关生物学过程的有20个，其中上调基因13个、下调基因7个。GO数据库分析显示，参与炎症相关的差异表达基因主要是CCR8、CCL19、CCL4、IL-17、IRF1、IFN-γ、SLC11A1等，涉及急性炎症反应、趋化因子受体活性、急性期反应、炎症反应和炎症反应的正调节等生物学过程。KEGG数据库分析显示，差异表达炎症相关基因主要富集在Toll样受体信号通路、NF-κB信号通路、IL-17信号通路、TNF信号通路和炎症性肠病等；实时荧光定量PCR结果显示，IFN-γ和MALT基因的相对表达倍数与转录组结果基本一致。本试验完成了DPV感染樱桃谷鸭十二指肠的转录组测序分析，获取了差异表达基因的功能注释信息，初步揭示了参与DPV感染的炎症相关通路，为深入探究DPV的致病机制提供了理论参考。

关键词: 鸭瘟病毒(DPV); RNA-seq; 十二指肠; 差异表达; 炎症相关基因; 调控通路

Abstract: This experiment was aimed to analyze the changes in the duodenal mucosal transcriptome level of duck plague virus (DPV) infected Cherry Valley ducks,and screen out genes and regulatory pathways involved in inflammation after DPV infection.DEV-GZ strain was used to inoculate 35 days old ducks with leg muscles,and the samples of duck duodenal mucosa tissues were collected at 24,48 and 72 h after inoculation,total RNA was extracted,and transcriptome sequencing was performed.Quality control and annotation of data were performed to screen differentially expressed genes and regulatory pathways related to inflammation,and some of the differentially expressed genes were verified by Real-time quantitative PCR.The results showed that a total of 21.39 Gb Clean Bases was obtained in control and experiment groups,the effective sequence of sequencing samples accounted for more than 99% of the original sequence,and the mapping rate was higher than 83%.There were 221 differentially expressed genes in duck duodenal mucosa after exposure 24 h with DEV-GZ,and 3 genes involved in inflammation-related biological processes,all of which were down-regulated genes.499 differentially expressed genes in duck duodenal mucosa after exposure 48 h with DEV,and 17 genes involved in inflammation-related biological processes,all of which were all down-regulated genes.798 differentially expressed genes in duck duodenal mucosa after exposure 72 h with DEV-GZ,and 20 were involved in inflammation-related biological processes,including 13 up-regulated genes and 7 down-regulated genes.GO database analysis showed that the differentially expressed genes involved in inflammation were mainly CCR8,CCL19,CCL4,IL-17,IRF1,IFN-γ,SLC11A1,etc.,involving acute inflammation,chemokine receptor activity,acute phase reaction and inflammation biological processes such as the positive regulation of reactions and inflammation.KEGG database analysis showed that differentially expressed inflammation-related genes were mainly enriched in Toll-like receptor signaling pathway,NF-κB signaling pathway,IL-17 signaling pathway,TNF signaling pathway and inflammatory bowel disease.Real-time quantitative PCR results showed IFN-γ and MALT genes were basically consistent with the transcriptome results.This experiment completed the transcriptome sequencing analysis of the duodenum of Cherry Valley duck infected by DPV,obtained the functional annotation information of differentially expressed genes,and initially revealed the inflammation-related pathways involved in DPV infection,provided a theoretical reference for further explore the pathogenic mechanism of DPV.

Key words: duck plague virus(DPV); RNA-seq; duodenum; differential expression; inflammation-related genes; regulatory pathways

中图分类号:

S855.3

田琴, 张明华, 周碧君, 程振涛, 王开功, 文明. 鸭瘟病毒感染鸭十二指肠黏膜炎症相关基因及调控通路的筛选[J]. 中国畜牧兽医, 2020, 47(10): 3323-3333.

TIAN Qin, ZHANG Minghua, ZHOU Bijun, CHENG Zhentao, WANG Kaigong, WEN Ming. Screening of Duodenal Mucosal Inflammation Related Genes and Regulatory Pathways in Ducks with Duck Plague Virus Infection[J]. China Animal Husbandry and Veterinary Medicine, 2020, 47(10): 3323-3333.

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