RNA-Seq-based transcriptomic profiling of primary interstitial cells of Cajal in response to bovine viral diarrhea virus infection
RNA-Seq-based transcriptomic profiling of primary interstitial cells of Cajal in response to bovine viral diarrhea virus infection
复制标题
基于 RNA-Seq 的 Cajal 原代间质细胞响应牛病毒性腹泻病毒感染的转录组分析
DOI：
10.1007/s11259-019-09754-y
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发表时间：
2019-08-01
影响因子：
2.2
通讯作者：
Shi, Huijun
中科院分区：
文献类型：
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作者：
Li, Shengnan;Hu, Xinyan;Shi, Huijun
牛病毒性腹泻病毒(BVDV)感染在牛肉和乳制品行业造成与健康相关的经济损失，并在世界各地广泛传播。严重的急性BVDV感染的特征是胃肠道(GI)炎症反应。BVDV引起炎性损害的机制尚不清楚。Cajal(ICC)网络的间质细胞在胃肠运动的电慢波的产生中起着关键的起搏器作用，对肠道神经系统的调节输入的接收是至关重要的。本研究探讨ICC是否是研究BVDV感染引起的胃肠道炎性损害的良好模型。从美利奴绵羊十二指肠分离出原代ICC。BVDV感染后培养5代的ICC中检测到BVDV的存在，表明BVDV在ICC中复制成功。感染BVDV TC株后，细胞增殖缓慢或下降。观察到ICC的形态变化，包括肿胀、溶解和形成空泡，表明细胞在数量、形态和功能上的变化。应用RNA测序技术(RNA-Seq)研究BVDV感染ICC的差异表达基因，探讨ICC数量、形态和功能改变的分子机制。BVDV感染后差异表达基因806个，其中538个基因表达上调，268个基因表达下调。基因本体论(GO)富集法和京都基因和基因组百科全书(KEGG)途径分析表明，806个DEG在27条途径中显著富含，包括细胞因子-细胞因子受体相互作用、白细胞介素17(IL-17)信号通路和丝裂原活化蛋白激酶(MAPK)信号通路。本研究高通量测序的DEGS和RAW文件已提交到NCBI基因表达总表(GEO)数据库(登录号：GSE122344)。最后，随机选择21个deg，用实时定量聚合酶链式反应(qRT-PCR)检测这些基因的相对抑制水平，以验证RNA-Seq结果。结果表明，21个DEGS的相关表达水平与RNA-Seq相似。本研究首次建立了一种新的研究BVDV感染引起的胃肠道炎性病变的模型。基于RNA-Seq的转录图谱可以为BVDV相关炎性病变的研究提供基础。
Infections with bovine viral diarrhea virus (BVDV) contribute significantly to health-related economic losses in the beef and dairy industries and are widespread throughout the world. Severe acute BVDV infection is characterized by a gastrointestinal (GI) inflammatory response. The mechanism of inflammatory lesions caused by BVDV remains unknown. The interstitial cells of Cajal (ICC) network plays a pivotal role as a pacemaker in the generation of electrical slow waves for GI motility, and it is crucial for the reception of regulatory inputs from the enteric nervous system. The present study investigated whether ICC were a good model for studying GI inflammatory lesions caused by BVDV infection. Primary ICC were isolated from the duodenum of Merino sheep. The presence of BVDV was detected in ICC grown for five passages after BVDV infection, indicating that BVDV successfully replicated in ICC. After infection with BVDV strain TC, the cell proliferation proceeded slowly or declined. Morphological changes, including swelling, dissolution, and formation of vacuoles in the ICC were observed, indicating quantitative, morphological and functional changes in the cells. RNA sequencing (RNA-Seq) was performed to investigate differentially expressed genes (DEGs) in BVDV-infected ICC and explore the molecular mechanism of underlying quantitative, morphological and functional changes of ICC. Eight hundred six genes were differentially expressed after BVDV infection, of which 538 genes were upregulated and 268 genes were downregulated. Gene Ontology (GO) enrichment and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses showed that the 806 DEGs were significantly enriched in 27 pathways, including cytokine-cytokine receptor interaction, interleukin (IL)-17 signaling and mitogen-activated protein kinase (MAPK) signaling pathways. The DEGs and raw files of high-throughput sequencing of this study were submitted to the NCBI Gene Expression Omnibus (GEO) database (accession number GSE122344). Finally, 21 DEGs were randomly selected, and the relative repression levels of these genes were tested using the quantitative real-time PCR (qRT-PCR) to validate the RNA-Seq results. The results showed that the related expression levels of 21 DEGs were similar to RNA-Seq. This study is the first to establish a new infection model for investigating GI inflammatory lesions induced by BVDV infection. RNA-Seq-based transcriptomic profiling can provide a basis for study on BVDV-associated inflammatory lesions.