• 大連大學(xué)醫(yī)學(xué)院檢驗(yàn)系（遼寧大連，116622）;

【摘要】　目的　 研究納米銀體外抗H3N2流感病毒的作用，并初步探索其作用機(jī)制。　方法　 在H3N2流感病毒吸附細(xì)胞后加入納米銀和吸附前用納米銀預(yù)處理犬腎細(xì)胞（MDCK），在體外用細(xì)胞病變效應(yīng)（cytopathic effect，CPE）觀察法和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）測值法，分析納米銀對H3N2流感病毒感染MDCK細(xì)胞的預(yù)防作用、直接滅活作用以及對流感病毒子代病毒體生成的抑制作用，運(yùn)用RT-PCR法研究納米銀對H3N2流感病毒HA基因復(fù)制的干擾作用。　結(jié)果　 納米銀能明顯殺傷H3N2流感病毒，50、25 μg/mL的納米銀溶液與H3N2流感病毒充分作用2 h后感染MDCK細(xì)胞，細(xì)胞存活率分別為94.38%和92.17%，納米銀能有效抑制流感病毒對MDCK細(xì)胞的侵入和侵入后病毒的繼續(xù)增殖，25 μg/mL納米銀溶液通過上述兩種方式處理細(xì)胞，細(xì)胞存活率分別為85.39%和83.28%，與病毒對照組相比，差異均有統(tǒng)計(jì)學(xué)意義（P lt;0.001）；400、200 μg/mL納米銀溶液分別與流感病毒H3N2充分混合作用15、30、60、120 min后，病毒液的HA基因均未能成功擴(kuò)增，純病毒液和溶劑對照組在1 700 bp處均出現(xiàn)明顯條帶。　結(jié)論　 通過3種不同的給藥方式，納米銀在體外均能明顯抑制流感病毒對細(xì)胞的感染，納米銀抑制流感病毒的機(jī)制可能是通過干擾H3N2流感病毒和吸附、穿入和基因的復(fù)制，從而抑制子代病毒體的生成。
【Abstract】　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 TCID50 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 lt;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.

引用本文： 向冬喜,蔣曉婷,李秀景,尹儉儉,陳娜娜,尹蕾,鄭叢龍. 納米銀體外抗H3N2流感病毒作用及其機(jī)制初步探討. 華西醫(yī)學(xué), 2011, 26(1): 4-7. doi: 復(fù)制

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  2. 2. 　Wang TT, Tan GS, Hai R, et al. Broadly protective monoclonal antibodies against H3 influenza viruses following sequential immunization with different hemagglutinins[J]. PLoS Pathog. 2010, 6(2): e1000796.
  3. 3. 　WHO. Influenza (Seasonal) Fact sheet 211[EB/OL]. [2009-04]. http://www.who.int/mediacentre/factsheets/fs211/en/.
  4. 4. 　Sun RW, Chen R, Chung NP, et al. Silver nanoparticles fabricated in Hepes buffer exhibit cytoprotective activities toward HIV-1 infected cells[J]. Chem Commun (Camb), 2005, (40): 5059-5061.
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  6. 6. 　Lu L, Sun RW, Chen R, et al. Silver nanopaarticles inhibit hepatitis B virus replication[J]. Antivir Ther, 2008, 13(2): 253-262.
  7. 7. 　向冬喜, 陳娜娜, 裘紅梅, 等. 納米銀毒理學(xué)安全性研究[J]. 實(shí)用醫(yī)學(xué)雜志, 2010, 26 (11): 1906-1908.
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