实验动物科学 ›› 2020, Vol. 37 ›› Issue (03): 1-.

• 研究报告 •    下一篇

CAV-1感染MDCK中各IFN及受体分子表达谱的差异

  

  1. (中国农业科学院哈尔滨兽医研究所实验动物与比较医学团队/兽医生物技术国家重点实验室, 黑龙江省实验动物与比较医学重点实验室,哈尔滨150069)
  • 出版日期:2020-06-28 发布日期:2020-10-26

Expression Levels of Various Interferon mRNA and Their Receptors in CAV-1-infected MDCK Cells

  • Online:2020-06-28 Published:2020-10-26

摘要: 目的 分析犬肾细胞系(MDCK)感染犬腺病毒1型(CAV-1)后各种干扰素(IFN)亚型及其受体分子转录水平的表达情况。方法 建立犬 IFN-β、IFN-β 受体(IFNAR1)、IFN-λ1、IFN-λ1 受体(IFN-λ1R)、IFN-λ3 和 IFN-λ3 受 体(IFN-λ3R)的染料法定量PCR方法,分析了 MDCK在Poly I:C处理和接种CAV-1后细胞内各IFN及受体分子转录水平的相对含量。结果 正常MDCK细胞内存在较高含量的IFN-X3R且含量显著高于其他IFN及受体分子的 转录水平;用Poly I:C处理MDCK细胞24 h后,IFN-X1R、IFN-X3和IFN-X3R的表达量均显著性上调,而IFN-λ1R、IFN-λ3和IFN-λ3R几乎没变化;接种CAV-1后,IFN-α的含量急剧上升至6倍。结论 MDCK细胞存在不同水平浓度的 IFN分子及其受体转录产物,且在Poly I:C刺激和腺病毒感染时受体分子的表达谱不同。

关键词: 犬肾细胞系, 干扰素受体, 转录表达水平

Abstract: Objective To understand the expression of IFNs and its receptor molecules in Madin-Darby Canine Kidney (MDCK) cell line infected with canine adenovirus type 1 ( CAV-1 ) . Method Several SYBR real-time quantitative PCR( RT-PCR) method were established to detect IFN-β, IFN-βreceptor. ( IFNAR1 ) IFN-λ1, IFN-λ1 receptor( IFN-λ1R) , IFN-λ3 and IFN-λ3 receptor( IFN-λ3R) in canine. The relative transcriptional levels of IFNs and its receptor molecules in Poly I: C treated-MDCK cells and CAV-1 treated-MDCK cells were analyzed. Result The transcriptional level of IFN-λ3R were significantly higher than that of other IFNs and receptor molecules in normal MDCK cells. The transcriptional levels of IFN-λ1R, IFN-λ3 , IFN-λ3R increased significantly in MDCK cells treated with Poly I: C for 24 h, while those of IFN-β, IFN-λ1, and IFNAR1 remained unchanged. However, the transcriptional level of IFN-α was the most highly than others in MDCK cells infected with CAV-1 for 24 h. Conclusion The result showed that there are different levels of various IFN molecules and their receptor transcripts in MDCK cells, and the express pattern varies after induction by. Poly I :C or adenovirus.

Key words: Madin-Darby Canine Kidney cells, Interferon receptors, transcriptional expression levels