多杀性巴氏杆菌攻毒小鼠肺脏的转录组分析

Transcriptomic Analysis of Lungs in Mice Challenged by Pasteurella multocida

多杀性巴氏杆菌(Pasteurella multocida)可感染多种动物宿主并引发败血症和呼吸道疾病,危害动物健康,给养殖业带来严重经济损失。本研究为探究多杀性巴氏杆菌感染期间对宿主肺脏基因表达的影响,选取羊源A型多杀性巴氏杆菌对小鼠(Mus musculus)进行攻毒实验,攻毒后72 h采集小鼠肺脏组织并提取总RNA。基于转录组测序,与对照组相比,攻毒组获得2443个差异表达基因,其中有1437个基因上调,1006个基因下调。对2443个差异表达基因进行GO富集分析,结果显示,差异基因显著富集的GO terms主要有B细胞稳态、T细胞迁移的正调控、整合素复合物和胶原蛋白结合;KEGG富集分析结果显示,差异基因显著富集于11条免疫炎症信号通路,包括细胞因子与细胞因子受体互作、趋化因子信号通路等;免疫炎症相关信号通路中,差异基因经蛋白质-蛋白质互作分析,结果显示,C3、C3ar1、C5ar1、Cxcl10、Cxcr2和Gng11基因处于网络中心,说明这些基因在多杀性巴氏杆菌感染过程中发挥了重要作用;从11条免疫炎症通路的基因中挑选10个进行实时荧光定量PCR验证,发现有8个基因表达趋势与测序结果相符,说明测序数据可靠。本研究展示了小鼠在多杀性巴氏杆菌感染过程中参与免疫反应的关键基因,为探索宿主抗多杀性巴氏杆菌感染分子机制提供了理论依据。

Pasteurella multocida(Pm)infects a variety of animals and causes host septicemic and respiratory diseases.It endangers animal health and leads to serious economic losses in livestock industry.To explore the effect of the gene expression of the host in the lungs during Pm infection,Pm serotype A was selected to cha-llenge mice,and murine lungs were collected 72 hours after challenging,of which total RNA was extracted.With transcriptome sequencing,2443 differentially expressed genes(DEGs)were found in challenged group compared with control group,including 1437 genes up-regulated and 1006 genes down-regulated.The results from GO enrichment analysis of 2443 DEGs showed that the significantly differential genes mainly were involved in B cell homeostasis,positive regulation of T cell migration,integrin complex and collagen binding;KEGG enrichment analysis of the source genes indicated that 11 immune and inflammation-related signal pathways including cytokine-cytokine receptor interaction,chemokine signaling pathway were significantly enriched;The protein-protein interaction analysis of the significantly differential genes in the 11 immune-inflammation-related signals pathways suggested that the C3,C3 ar1,C5 ar1,Cxcl10,Cxcr2 and Gng11 genes are in the center of the network,indicating that these genes play important roles during Pm infection;10 genes of 11 immune-inflammation-related signals pathways were selected to verified by RT-qPCR,and 8 genes were consistent with the RNA-seq results on expression trends,which indicating that the transcriptome sequencing data is reliable.This study revealed the critical genes in immune response of mice during Pm infection,which offered theory evidence for exploring the molecular mechanism of host defending against Pm attacks.