鸭肠炎病毒感染诱导鸭胚成纤维细胞γ干扰素上调表达及初步机制的研究

摘要

目的 探讨鸭肠炎病毒(DEV)诱导鸭胚成纤维细胞(DEF)中γ干扰素(IFNγ)的表达情况及初步机制.方法 分别提取感染DEV后24h、36h、48 h、60h的DEF总RNA,反转录成cDNA,使用特异性引物,运用荧光定量PCR方法,检测IFNγ及几个相关干扰素刺激基因(ISGs)mRNA水平的变化.结果 感染DEV的细胞相对于未感染的细胞,删的mRNA在24 h、36 h、48 h、60 h均显著上调,且相关ISGs包括抗黏液病毒基因(Mx)、DEAD框蛋白50(DDX50,是一种ATP依赖的RNA解旋酶)、2'-5'寡腺苷酸合成酶L(OASL)等基因的mRNA表达水平也均显著上调.结论 DEV感染DEF能够诱导IFNγ产生及相关ISGs表达的上调.%Objective In order to investigate the mechanism of interferon γ (IFNγ) up-regulation in duck embryo fibroblasts (DEF) induced by duck enteritis virus(DEV) infection.Methods The total RNA was extracted from DEF at 24 h,36 h,48 h and 60 h after DEV infection,and reverse transcripted into cDNA.The expression of IFNγ and related interferon stimulated genes (ISGs) was detected by fluorescence quantitative PCR using specific primers.Results The expression level of IFNγwas significantly up-regulated at 24 h,36 h,48 h and 60 h,and the mRNA expression levels of Myxovirus resistance (Mx)、DEAD-box protein 50(DDX50)、2'-5' oligoadenylates synthesis L (OASL) were also up-regulated in DEV-infected cells compared with those of uninfected cells.Conclusions DEV-infected DEF cells were able to induce IFNγ production and up-regulation of related ISGs.