摘要
旨在了解马疱疹病毒1型(EHV-1)导致脊髓型脑病的机制。采用EHV-1/China/YL2023株,以鼻内接种病毒的方式感染叙利亚金黄地鼠建立脑脊髓炎模型,并对脑组织进行转录组学测序分析。结果:感染EHV-1/China/YL2023分离株的叙利亚金黄地鼠,第5天出现食量减退、精神沉郁、口部流涎、皮毛凌乱等症状,且较对照组更具有攻击性;第7天开始出现神经症状,转圈、头点地即下垂,共济失调、站立不稳、行走困难、颈部僵硬,眼观发现脑充血、水肿以及轻度液化,对脑部进行苏木精伊红(HE)染色显示神经元肿胀、坏死及大量炎性细胞浸润;实时荧光定量PCR(RT-qPCR)检测发现促炎细胞因子γ干扰素(IFN-γ)与肿瘤坏死因子α(TNF-α)基因的表达量发生显著变化,表明EHV-1/China/YL2023分离株感染叙利亚金黄地鼠脊髓性脑病模型成功建立。转录组学测序分析表明,差异表达基因中有719个表达基因上调,173个基因表达量下调;基因本体(GO)富集分析表明,差异基因主要富集于免疫反应、免疫系统过程、防御反应、细胞增殖、胞外间隙、内吞囊泡、蛋白结合、信号受体结合、细胞因子受体结合、CXCR趋化因子受体结合;京都基因与基因组百科全书(KEGG)分析显示,主要富集的通路包括病毒蛋白与细胞因子及细胞因子受体的相互作用、抗原加工与提呈、肿瘤坏死因子(TNF)信号通路、趋化因子信号通路及NOD样受体信号通路、吞噬体信号通路、细胞黏附分子以及自然杀伤细胞介导的细胞毒性信号通路。因此,本研究构建了叙利亚金黄地鼠脊髓性脑病感染模型,对其进行转录组学测序分析,得到多个与EHV-1感染组织的差异表达基因和所富集的通路,为研究马鼻肺炎的分子机制提供基础,为后续的防控研究及新型疫苗提供新思路。
This study aims to understand the mechanism of myelopathic encephalopathy caused by equine herpesvirus type 1(EHV-1).The Syrian golden hamsters were infected with the virus strain EHV-1/China/YL2023 by intranasal inoculation to establish a myelopathic encephalopathy model,and the brain tissues were subjected to transcriptome sequencing analysis.The results were as follows:the Syrian golden hamsters infected with the EHV-1/China/YL2023 strain developed symptoms such as decreased food intake,depression,drooling,and disheveled fur on about the 5th day,and they were more aggressive than those hamsters of the control group.On about the 7th day,the hamsters began to develop neurological symptoms,turning and head drooping. The eye examination revealed cerebral hyperemia,edema andmild liquefaction. The hematoxylin-eosin ( HE) staining of the brain showed neuronal swelling,necrosis and a large number of inflammatorycells infiltration. Viral DNA could be detected by fluorescence quantitative PCR,and the expression of proinflammatory cytokine γ ( IFN-γ)and proinflammatory cytokine α ( TNF-α) genes was significantly changed,as shown by real-time fluorescent quantitative PCR. The resultsshowed that the Syrian golden hamster model of spinal encephalopathy infected with EHV-1 /China /YL2023 strain was successfully established.The transcriptome sequencing analysis showed that 719 genes were up-regulated and 173 genes were down-regulated in the differentiallyexpressed genes. The gene ontology ( GO) enrichment analysis showed that the differentially expressed genes were mainly enriched in immuneresponse,immune system process,defense response,cell proliferation,extracellular space,endocytic vesicles,protein binding,signalreceptor binding,cytokine receptor binding,and CXCR chemokine receptor binding. The Kyoto encyclopedia of genes and genomes ( KEGG)analysis showed that the main enriched pathways included the interaction of viral proteins with cytokines and cytokine receptors,antigen processingand presentation,tumor necrosis factor ( TNF) signaling pathway,chemokine signaling pathway and NOD-like receptor signalingpathway,phagosome signaling pathway,cell adhesion molecules and natural killer cell-mediated cytotoxicity signaling pathway. To sun up,in this study,a Syrian golden hamster spinal encephalopathy infection model was constructed,and transcriptome sequencing analysis was performedto obtain a number of differentially expressed genes and enriched pathways,compared with EHV-1 infected tissues;which provided abasis for study of the molecular mechanism of equine rhinorrhea and offered new ideas for future research on the prevention of the diseasesand development of new vaccines.
作者
撒瑞雪
杨一明
赵敏
贾晗铎
付玉
李银涛
张嗣玉
刘建华
冉多良
胡月
加尔肯
SA Ruixue;YANG Yiming;ZHAO Min;JIA Handuo;FU Yu;LI Yintao;ZHANG Siyu;LIU Jianhua;RAN Duoliang;HU Yue;Jiarken(Laboratory of Animal Infectious Diseases,College of Veterinary Medicine,Xinjiang Agricultural University,Urumqi 830052,China;Xinjiang Key Laboratory of New Drug Research and Development for Herbivores,College of Veterinary Medicine,Xinjiang Agricultural University,Urumqi 830052,China)
出处
《畜牧与兽医》
北大核心
2025年第3期95-102,共8页
Animal Husbandry & Veterinary Medicine
基金
中央引导地方科技发展专项资金项目(ZYYD2023C03)。
关键词
马疱疹病毒1型
差异表达基因
转录组
动物模型
equine herpesvirus type 1
differentially expressed gene
transcriptome
animal model
