禽呼肠病毒感染鸡胚成纤维细胞后 Toll 样受体mRNA 转录水平的动态变化

摘要

为分析禽呼肠病毒（ARV）标准毒株 S1133株感染鸡胚成纤维细胞（CEF）后对相关鸡 Toll 样受体（ChTLRs）mRNA 转录水平的影响作用，利用实时荧光定量 PCR，测定和分析 ARV-S1133感染 CEF后 ARV 结构蛋白σC 和 ChTLRs 的 mRNA 转录水平变化情况。结果表明，ARV-S1133感染 CEF 10 h 后， ARVσC 蛋白的 mRNA 相对表达量开始迅速上升，在48 h 达到峰值；同时，感染 CEF 中的 ChTLR3、ChTLR5、ChTLR7、ChTLR15和 ChTLR21 mRNA 表达量发生显著变化，在感染72 h 内各个受体的 mR-NA 表达量呈波浪式变化。5个不同滴度的 ARV 感染 CEF 24 h 后，ChTLR3、ChTLR5、ChTLR7、ChTLR15、ChTLR21 mRNA 转录水平与病毒滴度均呈正线性相关。上述结果表明，ARV 感染后可诱导CEF ChTLR3、ChTLR5、ChTLR7、ChTLR15、ChTLR21的 mRNA 转录水平发生变化，可能与禽呼肠病毒的复制和致病机制相关。%In order to analyze the mRNA transcriptic changes of chicken Toll-like receptors in ARV standard strain S1133-infected CEF cells,real-time PCR was used to analyze the mRNA transcriptic changes of ARV structural proteinσC and chicken Toll-like receptors in ARV S1133-infected CEF cells.The results showed that the relative expression levels of ARV structural proteinσC were elevated since 10 h post-infection,reaching a peak at 48 h post-infection.During this period,the mRNA transcriptic levels of ChTLR3,ChTLR5,ChTLR7,ChTLR15 and ChTLR21 genes were changed significantly in the infected CEF cells with different patterns.Meanwhile,the results of different ARV dose infection assay showed that the mRNA transcriptic levels of ChTLR3,ChTLR5,ChTLR7, ChTLR15and ChTLR21 genes were positively correlated with the virus doses.These data above suggest that ARV infection could effectively induce the mRNA expression changes of ChTLR3,ChTLR5,ChTLR7,ChTLR15 and ChTLR21.Furthermore,ChTLR3,ChTLR5,ChTLR7,ChTLR15 and ChTLR21 may play important roles in repli-cation and pathogenic mechanisms of ARV.