斑鳜肠道组织对嗜水气单胞菌感染早期的免疫响应  

Early immune response to Aeromonas hydrophila challenge in the intestinal tissue of Siniperca scherzeri

作　　者：赖铭勇[1] LAI Mingyong(Freshwater Fisheries Research Institute of Fujian Province,Fuzhou,Fujian 350002,China)

机构地区：[1]福建省淡水水产研究所,福建福州350002

出　　处：《福建农林大学学报（自然科学版）》2024年第6期763-771,共9页Journal of Fujian Agriculture and Forestry University:Natural Science Edition

基　　金：福建省种业创新与产业化工程渔业项目(闽农宗[2021]5号);国家现代农业产业技术体系项目(CARS-46)。

摘　　要：【目的】嗜水气单胞菌是危害斑鳜养殖生产的主要病原之一。研究斑鳜感染嗜水气单胞菌后,其肠道组织基因表达水平的变化,解析鱼体对细菌性感染的免疫应答分子机制,为防控淡水鱼类细菌性疾病提供依据。【方法】用嗜水气单胞菌感染斑鳜6 h后,利用Illumina Hiseq 6000测序平台对斑鳜肠道组织进行RNA-seq测序,并利用生物信息学方法对数据进行分析。【结果】感染组和未感染组斑鳜肠道组织存在1077个差异表达基因(DEGs),包括669个上调DEGs和408个下调DEGs。其中,与免疫相关的上调DEGs主要有基质金属蛋白酶3(MMP3)、白细胞介素-1β(IL-1β)、白细胞介素-17F(IL-17F)、正五聚蛋白3(PTX3)、核苷酸结合寡聚化结构域样受体(NLR)含pyrin结构域蛋白6(NLRP6)、C型凝集素域家族4E(CLEC4E)、胆固醇-25-羟化酶(CH25H)和激活转录因子1(ATF1)等。基因本体(GO)富集分析显示,DEGs主要富集在免疫反应激活信号通路、免疫应答调节细胞表面受体信号通路、T细胞活化的正调控过程以及细胞修复和调节过程上。京都基因与基因组百科全书(KEGG)富集分析显示,上调DEGs主要富集在多种疾病发生、发展过程和免疫代谢相关信号通路上。实时荧光定量PCR验证结果表明,7个随机选取的与免疫相关的DEGs的表达量与RNA-seq测序结果存在一致性。【结论】感染组与未感染组斑鳜肠道组织MMP3、IL-1β、PTX3、NLRP6、CLEC4E表达量的差异较为显著,可能是参与斑鳜肠炎发生的关键候选基因。【Objective】Aeromonas hydrophila is one of the main pathogens severely endangering Siniperca scherzeri farming and production.Level of intestinal gene expression of S.scherzeri was analyzed after A.hydrophila infection to explore the molecular defense mechanism of S.scherzeri against bacterial infection,thus to provide basis for its control.【Method】After infection with A.hydrophila for 6 h,transcriptome sequencing and bioinformatics analysis were performed on intestinal tissue of S.scherzeri using Illumina Hiseq 6000 platform.【Result】A total of 1077 differentially expressed genes(DEGs)were screened between the infected group and control group,including 669 up-regulated and 408 down-regulated genes.Among them,the up-regulated genes related to immune response contained matrix metalloproteinase-3(MMP3),interleukin-1β(IL-1β),interleukin-17F(IL-17F),pentraxin-3(PTX3),nucleotide binding oligomerization domain like receptor(NLR)family pyrin domain containing 6(NLRP6),C-type lectin domain family 4 member E(CLEC4E),cholesterol 25-hydroxylase(CH25H),and activating transcription factor 1(ATF1).Gene ontology(GO)analysis revealed that the DEGs were mainly involved in the immune response-activating signaling pathway,immune response-regulating cell surface receptor signaling pathway,positive regulation of T cell activation,and cell repair and regulation.KEGG enrichment analysis showed that the up-regulated DEGs were mainly enriched in the occurrence and development process of various diseases,and immunity-related metabolic pathways.RT-qPCR showed consistent result with RNA-seq analysis on 7 DEGs.【Conclusion】MMP3,IL-1β,PTX3,NLRP6 and CLEC4E,which were significantly differentially expressed between the infected group and control group,may be key the genes involved in the intestinal inflammation of S.scherzeri.

关 键 词：斑鳜 嗜水气单胞菌 转录组 差异表达基因 

分 类 号：S941.42[农业科学—水产养殖]