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为深入研究牛场中普遍存在的持续性感染,探讨不同生物型牛病毒性腹泻病毒(bovine viral diarrhea virus,BVDV)转化的分子机制,本试验将致细胞病变型BVDV(CP型BVDV)和非致细胞病变型BVDV(NCP型BVDV)感染MDBK细胞,观察细胞变化,感染后12~72h期间每间隔12h收集1次细胞,同时对24、48h收集的细胞进行转录组学分析。结果显示,不同生物型BVDV感染宿主细胞24h后,与感染NCP型BVDV的细胞相比,感染CP型BVDV的MDBK细胞中上调差异表达的基因2 849个,下调差异表达的基因3 347个,48h后,上调差异表达的基因2 933个,下调差异表达的基因2 831个。对差异基因的GO功能分析结果表明,差异基因参与的分子功能主要有催化活性、结合活性、酶调节活性、分子转导活性和蛋白结合转录因子活性等;Pathway显著性富集分析结果显示,差异表达基因主要参与细胞自噬、细胞凋亡、免疫调节因子等相关的信号通路。本试验结果可为进一步揭示病毒致病的分子机制、控制BVDV和研制其候选疫苗奠定基础。
In order to study the persistent infection prevailing in cattle farms and explore the molecular mechanism of the transformation of different biotype bovine viral diarrhea virus (BVDV) MDCK cells were infected with BVDV (NCP type BVDV). The changes of cells were observed. Cells were harvested at 12h intervals from 12 to 72h after infection. Transcriptome analysis was also performed on cells collected at 24 and 48h. The results showed that compared with those infected with NCP BVDV, the number of differentially expressed genes in MDBK cells infected with BVDV of CP type was 2 849, down-regulated 3 347, 48 h Afterwards, 2 933 genes were differentially expressed and 2 831 genes differentially expressed were down-regulated. The analysis of GO function of the differentially expressed genes showed that the molecular functions of the differentially expressed genes mainly include catalytic activity, binding activity, enzyme regulating activity, molecular transduction activity, and protein binding transcription factor activity. Significant enrichment analysis by Pathway showed that the difference Expression of genes involved in cell autophagy, apoptosis, immune regulatory factors and other related signaling pathways. The results of this study may lay the foundation for further revealing the molecular mechanism of virus pathogenesis, controlling BVDV and developing its candidate vaccine.