O型口蹄疫兔化弱毒YNTBa株感染的牛原代睾丸细胞基因表达分析  

Gene expression in primary bovine testicular cells infected with a foot-and-mouth disease virus serotype O rabbit-attenuated YNTBa strain

作　　者：汤雅淇 普娅芸 李珂 高华峰[2] 何于雯 王生奎 信爱国[2] TANG Yaqi;PU Yayun;LI Ke;GAO Huafeng;HE Yuwen;WANG Shengkui;XIN Aiguo(College of Veterinary Medicine,Yunnan Agricultural University,Kunming 650224,China;National Foot-and-mouth Disease Para-reference(Kunming),Yunnan Institute of Animal Science and Veterinary Medicine/Yunnan Tropical and Subtropical Animal Virus Disease Laboratory,Kunming 650224,China)

机构地区：[1]云南农业大学动物医学院,云南昆明650224 [2]云南省畜牧兽医科学院国家口蹄疫专业实验室(昆明)/云南省热带亚热带动物病毒病重点实验室,云南昆明650224

出　　处：《畜牧与兽医》2022年第10期89-96,共8页Animal Husbandry & Veterinary Medicine

基　　金：国家重点研发计划(2021YFD1800301);国家自然科学基金(31760745)。

摘　　要：为分析口蹄疫病毒(foot-and-mouth disease virus,FMDV)减毒株感染天然宿主细胞后基因表达变化,将O型FMDV兔化弱毒株YNTBa接种初生犊牛原代睾丸细胞(BT),采用二代测序技术对YNTBa感染BT细胞后8 h和24 h的细胞转录组进行研究。结果显示:病毒感染8 h和24 h的试验组较对照组分别筛选出437和134个表达变化≥2倍的差异表达基因。基因本体论(GO)功能类聚分析显示,差异表达基因主要参与到细胞分化、DNA转录、细胞因子产生和凋亡等。京都基因与基因组百科全书(KEGG)功能富聚分析表明,差异表达基因主要与细胞代谢、核糖体、吞噬体、蛋白消化吸收、细胞分化、细胞转录调控等相关,涉及糖基化终末产物受体(AGE-RAGE)、肿瘤坏死因子(TNF)、白细胞介素17(IL-17)、转化生长因子β(TGF-β)、磷脂酰肌醇-3-激酶/蛋白激酶B(PI3K-Akt)、丝裂原活化蛋白激酶(MAPK)、促性腺激素释放激素(GnRH)、雌性激素(estrogen)和催产素(oxytocin)等多条信号通路。利用实时荧光定量PCR(RT-qPCR)验证了其中12个差异表达基因,定量结果与转录组测序结果一致。本研究筛选的差异表达基因为进一步了解FMDV弱毒免疫机制,包括FMDV复制相关基因提供了基础。This study was to investigate the differentially expressed genes(DEGs)of natural host cells infected with an attenuated footand-mouth disease virus(FMDV).A transcriptome analysis of primary bovine testicular(BT)cells infected with an FMDV serotype O rabbit-attenuated YNTBa strain at 8 and 24 hours post infection(h.p.i.)was performed using the next-generation sequencing technique.During infection with YNTBa at 8 h.p.i and 24 h.p.i,a total of 437 and 134 DEGs that exhibited a≥2-fold change in expression were identified,respectively.The gene ontology(GO)annotation indicated that the DEGs were mainly involved in cell differentiation,DNA transcription,cytokine production and apoptosis.The Kyoto encyclopedia of genes and genomes(KEGG)enrichment analysis showed that the DEGs were primarily related to cell metabolism,ribosomes,phagocytes,protein digestion and absorption,cell differentiation,and cell transcriptional regulation.And the DEGs involved several signaling pathways,including AGE-RAGE,TNF,IL-17,TGF-β,PI3K-Akt,MAPK,GnRH,estrogen and oxytocin.Twelve DEGs were selected and validated using RT-qPCR,which showed that the expression patterns of these genes were consistent with the results of the transcriptome sequencing.This study presented the first transcriptome analysis of BT cells infected with a live-attenuated FMDV,and the results provided a basis for further understanding of the host genes involved in virus replication and immune mechanism of the live-attenuated FMDV.The DEGs screened in this study provided a basis for further research on the immune mechanism of FMDV attenuated viruses,including the genes related to FMDV replication.

关 键 词：O型口蹄疫病毒 减毒株 转录组分析 差异表达基因 

分 类 号：S852.6[农业科学—基础兽医学]