牛结节性皮肤病病毒感染MDBK细胞后转录组差异表达分析

doi:10.11843/j.issn.0366-6964.2021.02.018

畜牧兽医学报 ›› 2021, Vol. 52 ›› Issue (2): 460-468.doi: 10.11843/j.issn.0366-6964.2021.02.018

牛结节性皮肤病病毒感染MDBK细胞后转录组差异表达分析

曹政¹, 居马别克·夏拉巴依², 李春燕³, 马力克·艾则孜^2*

1. 重庆市畜牧科学院, 重庆 402460;
2. 新疆维吾尔自治区动物卫生监督所, 乌鲁木齐 830011;
3. 陆军军医大学第一附属医院全军临床病理学研究所, 重庆 400000

收稿日期:2020-08-10 出版日期:2021-02-23 发布日期:2021-02-24
通讯作者: 马力克·艾则孜,主要从事人畜共患病防控及动物寄生虫病研究,E-mail:2643094852@qq.com
作者简介:曹政(1984-),男,内蒙古正镶白旗人,博士,主要从事人畜共患病防控研究,E-mail:caozheng.1@gmail.com
基金资助:
科研机构绩效引导专项（CSSA2019ZN038）

Transcriptome Analysis of MDBK Cells Infected with Lumpy Skin Disease Virus

CAO Zheng¹, JUMABIEKE Xialabayi², LI Chunyan³, MALIKE Aizezi^2*

1. Chongqing Academy of Animal Sciences, Chongqing 402460, China;
2. Animal Health Supervision Institute of Xinjiang Uygur Autonomous Region, Urumqi 830011, China;
3. Institute of Clinical Pathology, First Affiliated Hospital of Army Medical University, Chongqing 400000, China

Received:2020-08-10 Online:2021-02-23 Published:2021-02-24

摘要/Abstract

摘要： 旨在探索牛结节性皮肤病病毒（lumpy skin disease virus，LSDV）与宿主细胞的相互作用，采集了具有典型临床症状的牛皮肤病变组织，接种MDBK细胞进行病毒分离，采用透射电镜观察病变组织及病毒感染后的MDBK细胞，同时对感染后24 h的MDBK细胞进行转录组学分析，筛选差异表达基因并将其按功能分类，分析它们参与的细胞信号通路。结果显示，LSDV在电镜下呈椭圆形，病毒粒子大小约320 nm×260 nm。转录组分析发现，与未处理组相比，显著差异表达的基因共499个，其中，112个基因上调表达，387个基因下调表达。通过GO和KEGG分析得知，差异表达基因主要富集在从细胞表面传导信号至细胞核内部的MAPK信号通路，其中，参与天然免疫且涉及细胞凋亡的MAP3K1基因表达呈现显著下调，提示病毒感染会影响宿主细胞的迁移与凋亡等过程。显著变化基因包括HIF1A、RORA、LRRK2、AP3B1和ANO6等，分别涉及到调节血管内皮生长因子、炎症反应、细胞增殖分化以及信号转导等功能，除此之外，不饱和脂肪酸及胆汁分泌两个信号通路也发生了显著变化。本研究为进一步深入探究LSDV的致病机制奠定了基础。

关键词: 牛结节性皮肤病病毒, MDBK细胞, 高通量测序, 转录组

Abstract: To explore the interaction between LSDV and host cells, in this study, cattle skin samples with typical lesions of LSD was collected, and MDBK cells were inoculated for virus isolation. Skin lesions and infected MDBK cells were observed by transmission electron microscopy. Besides, transcriptomics analysis was performed on MDBK cells 24 hours after infection to screen differentially expressed genes, classify them according to their functions, and analyze the cell signaling pathways involving them. The results showed that LSDV was elliptic under electron microscope and the virion size was about 320 nm×260 nm. Compared with the untreated group, a total of 499 genes were significantly differentially expressed, among which 112 genes were up-regulated and 387 genes were down-regulated through transcriptome analysis. According to GO and KEGG analysis, the differentially expressed genes were mainly concentrated in the MAPK signaling pathway that transmits signals from the cell surface to the inside of the nucleus, in which the expression of MAP3K1 gene, which is involved in innate immunity and associated in apoptosis, showed significant downregulation, suggesting that this viral infection may affect the migration and apoptosis of host cells. The significantly changed genes include HIF1A, RORA, LRRK2, AP3B1 and ANO6, which are respectively involved in functions such as regulating vascular endothelial growth factor (VEGF), inflammatory response, proliferation and differentiation of cells and signal transduction. This study laid a foundation for further exploring the pathogenic mechanism of LSDV.

Key words: lumpy skin disease virus (LSDV), MDBK cells, high throughput sequencing, transcriptome

中图分类号:

S852.659.1

曹政, 居马别克·夏拉巴依, 李春燕, 马力克·艾则孜. 牛结节性皮肤病病毒感染MDBK细胞后转录组差异表达分析[J]. 畜牧兽医学报, 2021, 52(2): 460-468.

CAO Zheng, JUMABIEKE Xialabayi, LI Chunyan, MALIKE Aizezi. Transcriptome Analysis of MDBK Cells Infected with Lumpy Skin Disease Virus[J]. Acta Veterinaria et Zootechnica Sinica, 2021, 52(2): 460-468.

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牛结节性皮肤病病毒感染MDBK细胞后转录组差异表达分析

Transcriptome Analysis of MDBK Cells Infected with Lumpy Skin Disease Virus

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