摘要
【目的】了解鸡肝癌细胞(LMH)感染血清4型禽腺病毒(FAdV-4)后干扰素(IFN)和干扰素刺激基因(ISGs)的表达量变化,为研究FAdV-4与ISGs的相互作用提供参考依据。【方法】试验将FAdV-4感染LMH细胞,观察不同时间点细胞病变,并收集感染0、12、24、36、48、60、72、84和96 h的细胞样品,通过实时荧光定量PCR技术检测感染病毒后不同时间点IFN-α和IFN-β及ISGs基因在转录水平上表达量的动态变化规律。【结果】LMH细胞感染FAdV-448 h时出现典型的细胞病变;在感染12 h后,FAdV-4快速增殖,60 h时达到峰值;感染病毒后各时间点,与对照组相比,IFN-α基因转录水平表达量均显著下调(P<0.05);与对照组相比,在感染后12和24 h,IFN-β基因表达量显著下调(P<0.05),在36 h时迅速上调表达至峰值(P<0.05),在48 h时仍维持在较高水平,之后时间点的表达量有所下降且趋于稳定,但仍显著高于对照组(P<0.05);ISG 12、IFIT 5及DDIT 4基因在感染前期变化不大,在感染后36 h时迅速上调,在感染后96 h达到峰值(P<0.05);ZFP 313、IFITM 3及Viperin基因在感染前期表达量变化不大,随后在36 h迅速上调表达至峰值(P<0.05),在48 h时仍维持在较高水平,之后时间点的表达量有所下降,但仍显著高于对照组(P<0.05);与对照组相比,CD 47、OAS和PKR基因均呈现下调表达。【结论】在FAdV-4感染LMH细胞后,IFN及多种ISGs在转录水平呈现规律性变化,与病毒在LMH细胞中复制存在一定的联系,表明这些天然免疫因子可能在抗FAdV-4反应中发挥着重要作用。
【Objective】This study was aimed to investigate the changes of interferon(IFN)and interferon-stimulating genes(ISGs)expression in chicken Leghorn male hepatoma(LMH)infected with Fowl adenovirus serotype-4(FAdV-4),and provide reference basis for the study of the interaction between FAdV-4 and ISGs.【Method】The experiment was conducted by infecting LMH cells with FAdV-4,observing the cellular lesions and collecting cell samples at 0,12,24,36,48,60,72,84 and 96 h post-infection,and the dynamic change patterns of the expression of IFN-αand IFN-βand ISGs genes at the transcriptional level at different time points after infection were detected by Real-time quantitative PCR.【Result】The results showed that after FAdV-4 infection of LMH cells,LMH cells showed typical cytopathic lesions at 48 h.It was detected that FAdV-4 proliferated rapidly at 12 h,and peaked at 60 h.Compared with control group,the expression of IFN-αat the transcriptional level was significantly down-regulated after infection(P<0.05).Compared with control group,the expression of IFN-βwas significantly down-regulated at 12 and 24 h after infection(P<0.05),and then rapidly and significantly up-regulated to the peak at 36 h(P<0.05),and remained at a high level at 48 h.After that,the expression declined and stabilized,but was still significantly up-regulated compared with control group(P<0.05).The expressions of ISG 12,IFIT 5 and DDIT 4 genes did not change much in the early stage of infection,and reached the peak at 96 h after infection(P<0.05).ZFP 313,IFITM 3 and Viperin genes did not change much in the early stage of infection,and then rapidly and up-regulated to peak at 36 h(P<0.05),and remained at a high level at 48 h.After that,the expression declined,but was still significantly up-regulated compared with control group(P<0.05).CD 47,OAS and PKR genes showed down-regulated expression compared with control group.【Conclusion】After FAdV-4 infection of LMH cells,IFN and a variety of ISGs showed regular changes at the transcriptional level
作者
万丽军
王盛
谢芝勋
任红玉
谢丽基
范晴
罗思思
李孟
张艳芳
曾婷婷
黄娇玲
张民秀
谢志勤
李小凤
韦悠
WAN Lijun;WANG Sheng;XIE Zhixun;REN Hongyu;XIE Liji;FAN Qing;LUO Sisi;LI Meng;ZHANG Yanfang;ZENG Tingting;HUANG Jiaoling;ZHANG Minxiu;XIE Zhiqin;LI Xiaofeng;WEI You(Guangxi Key Laboratory of Veterinary Biotechnology,Guangxi Veterinary Research Institute,Nanning 530001,China)
出处
《中国畜牧兽医》
CAS
CSCD
北大核心
2024年第5期2081-2090,共10页
China Animal Husbandry & Veterinary Medicine
基金
广西科技重大专项项目(桂科AA23062050)
“广西八桂学者”专项(2019A50)。
