脱氧核酶在培养人气道上皮细胞体系内的抗甲型流感病毒NS基因效应

Inhibition of influenza A virus in cultured 9HTEo cells by NS gene targeted DNAzyme

目的探讨针对甲型病毒(influenza Avirus,IFV)NS基因特异性的脱氧核酶(DNAzyme,DZ-NS)在培养的人气道上皮细胞(9HTEo)体系内对甲型流感病毒复制的抑制效应.方法光镜观察DZ-NS对IFV致9HTEo细胞病变效应变化的影响;病毒空斑形成阻断试验及四甲基偶氮唑盐比色试验(MTT)检测脱氧核酶DZ-NS对IFV复制的抑制和对感染IFV的9HTEo细胞的保护作用;RT-PCR检测DZ-NS对IFV mRNA表达的抑制作用。结果DZ-NS可明显改善IFV所致的细胞病变并能明显提高细胞感染IFV后的存活率(P<0.01);可显著抑制IFV-NS基因mRNA的表达;空斑实验病毒抑制率可达80.11%;DZ-NS在细胞内对IFA病毒表达的抑制效率,明显高于作为对照的反义寡核苷酸(antisense oligonucleotides,AS-NS)(P<0.05)。结论在9HTEo感染IFV细胞模型系统,DZ-NS能高效阻断IFV的NS基因的表达,是一种特异性的、高效的抗IFV基因治疗剂。

Objective To explore the inhibitory effects of specific DNAzyme targeting influenza A virus(IFV)NS gene(DZ-NS)on viral replication of IFV in 9HTEo cell culture system.Methods The effects of DZ-NS on cytopathogenic effect(CPE)of 9HTEo cell induced by IFV infection were observed microscopically.Viral plaque forming assay and MTT assay were used to detect the viral titer changing and protective function of the DZ-NS on IFV-infected 9HTEo cel1s,respectively.Inhibitory effects of DZ-NS on viral replication was d...