硫代磷酸反义寡核苷酸prop5对流感病毒进入细胞的抑制作用  

Inhibited effect of phosphorothioate antisense oligonucleotides prop5 on entry of influenza virus into cells

作　　者：李康[1,2] 齐向云[2] 李庆平[2] 夏燕平[2] 杨静[2] 王升启[2] 

机构地区：[1]北京大学生命科学与生物工程学院,北京100124 [2]军事医学科学院放射与辐射医学研究所,北京100850

出　　处：《国际药学研究杂志》2014年第1期102-107,117,共7页Journal of International Pharmaceutical Research

基　　金：国家自然科学基金面上项目(31270197;81230089)

摘　　要：目的研究硫代磷酸反义寡核苷酸prop5对流感病毒感染进入细胞的影响。方法采用流感病毒感染A549细胞模型,设立随机序列(prop5R)和正义序列(prop5S)作为阴性对照,收集细胞培养液,采用实时荧光定量PCR检测病毒RNA拷贝数,评价反义寡核苷酸对病毒吸附和进入的影响;进一步通过血凝抑制实验检测prop5对流感病毒吸附细胞的影响,溶血抑制实验检测prop5对流感病毒进入细胞的影响。结果 prop5能够抑制流感病毒A/Jingfang/1/86(H1N1)感染A549细胞,对流感病毒吸附细胞过程没有影响,prop5能够抑制A/Jingfang/1/86(H1N1)、A/Lufang/9/93(H3N2)、A/FM/1/47(H1N1)和A/PR/8/34(H1N1)介导的膜融合。结论 prop5能抑制流感病毒进入细胞,这种额外的活性增强了prop5的抗病毒能力。Objective To study the inhibitory acitvity of phosphorothioate antisense oligonucleotide （P-ASODN） prop5 against influenza virus entry into cells. Methods A549 cells infected by influenza virus were used in this study. A random sequence （prop5R） and a sense sequence （prop5S） served as negative controls. Culture media supernatants were collected to detect copies of viral RNA by real-time quantitative RT-PCR. Further, the effect of prop5 on virus attachment was validated by hemagglutination inhibition assay and its effect on virus entry was determined by hemolysis inhibition assay. Rusults Prop5 could inhibit infection of A549 by influenza A/Jingfang/1/86（H1N1）. Prop5 did not interfere with influenza virus attachment on cells. Prop5 could inhibit membrane fusion mediated by influenza A/Jingfang/1/86（H1N1）, A/Lufang/9/93（H3N2）, A/FM/1/47（H1N1） and A/PR/8/34（H1N1）. Conclusion Prop5 blocks influenza virus entry in sequence-independent manner. This extra activity enhances the anti-influenza virus capability of prop5.

关 键 词：硫代磷酸反义寡核苷酸 prop5 流感病毒 感染 

分 类 号：R97[医药卫生—药品]