纳米银体外抗H3N2流感病毒作用及其机制初步探讨  

Anti-viral Effects of Silver-nanoparticles on H3N2 Influenza Virus in vitro and Its Mechanism

作　　者：向冬喜[1] 蒋晓婷[1] 李秀景[1] 尹俭俭[1] 陈娜娜[1] 尹蕾[1] 郑丛龙[1] 

机构地区：[1]大连大学医学院检验系,辽宁大连116622

出　　处：《华西医学》2011年第1期4-7,共4页West China Medical Journal

基　　金：国家自然科学基金资助项目(30870681)~~

摘　　要：目的研究纳米银体外抗H3N2流感病毒的作用,并初步探索其作用机制。方法在H3N2流感病毒吸附细胞后加入纳米银和吸附前用纳米银预处理犬肾细胞(MDCK),在体外用细胞病变效应(cytopathic effect,CPE)观察法和3-(4,5-二甲基-2-噻唑)-2,5-二苯基溴化四唑(3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide,MTT)测值法,分析纳米银对H3N2流感病毒感染MDCK细胞的预防作用、直接灭活作用以及对流感病毒子代病毒体生成的抑制作用,运用RT-PCR法研究纳米银对H3N2流感病毒HA基因复制的干扰作用。结果纳米银能明显杀伤H3N2流感病毒,50、25μg/mL的纳米银溶液与H3N2流感病毒充分作用2h后感染MDCK细胞,细胞存活率分别为94.38%和92.17%,纳米银能有效抑制流感病毒对MDCK细胞的侵入和侵入后病毒的继续增殖,25μg/mL纳米银溶液通过上述两种方式处理细胞,细胞存活率分别为85.39%和83.28%,与病毒对照组相比,差异均有统计学意义(P<0.001);400、200μg/mL纳米银溶液分别与流感病毒H3N2充分混合作用15、30、60、120min后,病毒液的HA基因均未能成功扩增,纯病毒液和溶剂对照组在1 700bp处均出现明显条带。结论通过3种不同的给药方式,纳米银在体外均能明显抑制流感病毒对细胞的感染,纳米银抑制流感病毒的机制可能是通过干扰H3N2流感病毒和吸附、穿入和基因的复制,从而抑制子代病毒体的生成。Objective To explore the anti-viral effects of silver-nanoparticles（silver-nps） on H3N2 influenza virus in vitro and to evaluate its mechanism.Methods Silver-nps was added to canine kidney cells（MDCK） before and after the cells was adsorpted by H3N2 influenza virus.Cytopathic effect（CPE） assay and the 3-（4,5-dimethylthiazol-2-yl）-2,5-diphenyltetrazolium bromide（MTT） assay were used to analyze the preventive effect,directly off deactivation,and the inhibit formation of progeny virions of silver-nps on H3N2 viruses.The interference of HA gene replication was observed by the RT-PCR assay.Results The survival rate of MDCK cells was 94.38% and 92.17% after 50 and 25 μg/mL silver-nps were mixed with 100 TCID50 H3N2 virus in 2 hours,and the survival rate of MDCK cells was 85.39% and 83.28% before and after the cells was adsorpted by H3N2 influenza virus when 25 μg/mL silver-nps was added to the cells（all compared to virus control,P〈0.001）,which showed that silver-nps could inactivate H3N2 virus,prevente them invasing to the cells and reproducting when H3N2 entered the cell remarkedly.The HA gene was not amplified successfully when 50 and 25 μg/mL silver-nps were mixed with 100TCID50 H3N2 virus in 15,30,60,and 120 minutes later,but both pure virus solution and solvent control group appeared a significant bright band in the 1 700 bp area.Conclusion Under three different administration modes,silver-nps has an obvious effect against H3N2 in vitro,which could interfere the HA gene replication and inhibit the formation of H3N2 progeny virions.

关 键 词：纳米银 H3N2流感病毒 抗病毒作用 作用机制 

分 类 号：R319[医药卫生—基础医学]