纳米银体外抗腺病毒作用及机制研究  

Inhibitory effects of silver nanoparticles and their possible mechanism on adenovirus in vitro

作　　者：陈娜娜[1] 王云华[1] 尹俭俭[1] 李秀景[1] 郑丛龙[1] 

机构地区：[1]大连大学医学院病原生物学教研室,辽宁大连116622

出　　处：《大连医科大学学报》2011年第5期415-420,共6页Journal of Dalian Medical University

基　　金：国家自然科学基金面项目(30870681)

摘　　要：[目的]探讨纳米银(silver nanoparticles,silver-nps)在体外抗腺病毒3型(adenovirus type 3,ADV3)的作用机制。[方法]采用细胞培养方法和免疫荧光方法,观察纳米银对ADV3子代病毒体生成的抑制作用、对ADV3侵入细胞的阻断作用及直接灭活作用,同时用透射电镜、PCR方法观察体外纳米银对ADV3病毒体及其DNA的破坏作用。[结果]纳米银在Hela细胞上最大无毒浓度(TC0)为52.48μg/mL;ADV3在Hela细胞上的半数感染浓度(TCID50)为10-2.74/100μL;最大无毒剂量范围内,50μg/mL的纳米银和等体积100TCID50ADV3分别以杀病毒、抗病毒、直接灭活病毒三种不同方式作用Hela细胞,细胞存活率分别为(93.58±1.79)%,(90.50±1.99)%和(95.38±2.98)%,而100TCID50 ADV3感染Hela细胞后,测得细胞存活率为(31.92±2.86)%,两者相比差异有显著性意义(P<0.01)。免疫荧光显示,与腺病毒对照组形成的强特异性荧光相比,三种途径作用于Hela细胞后各组的特异性荧光颗粒均减少;电镜显示,纳米银可直接与病毒粒子相互作用,破坏ADV3粒子呈时间依赖性。PCR显示,与病毒对照组、溶剂对照组相比,纳米银组扩增目的条带亮度都相对较弱。[结论]纳米银在体外对ADV3具有明显的抑制作用,其作用机制可能与纳米银破坏病毒粒子及其表面衣壳蛋白及腺病毒DNA有关。[ Objective ] To explore the inhibitory effects and possible mechanism of silver nanoparticles （ silver - nps） on adenovirus type 3 （ ADV3 ）. [ Methods] Cell culture and immuno fluorescence assay were applied to evaluate the anti - adenovirus activities of silver - nps in inhibiting the formation of ADV3 progeny virions, blocking and inactivating ADV3. The anti - adenovirus activities of silver - nps on Hela cells were observed by transmission electron microscopy. The PCR method was also used to analyze the destructive effect of silver - nps on ADV3 DNA. [ Results ] TCo of silver - nps was 52.48 μg/ mL. The median tissue culture infective dose（ TCID50 ） of ADV3 on Hela cell was l0^-2.74/100 μL. In the range of non -toxic concentration, the 50 μg/mL of silver -nps and 100TCIDs0 of ADV3 were analyzed in virucidal, antiviral and inactivating ways, the survival rates of Hela cells were （ 93.58 ± 1.7 9 ） %, （ 90.50 ± 1.99 ） % and （ 95.38 ±2.98 ） % respectively, when compared with the viability of control group, the differences were significant （ P 〈 0.05 ）. Hela cells infected with ADV3 had much specific immunofluorescenee when compared to the cells treated with silver - nps in virucidal, antiviral and inactivating ways. Compared to the intact shape of ADV3 virions, the silver -nps could directly interact with viral particles and damage ADV3 virions in a time dependent manner under transmission electron microscopy. The densities of DNAbands of virions treated with silver -nps were decreased in comparison to that of control group following PCR amplification. [ Conclusion] Silvernps have inhibitory effects on ADV3 in vitro,its possible mechanisms are associated with damaging viral particles, capsid protein and DNA.

关 键 词：纳米银 腺病毒3型 抗病毒 

分 类 号：R373[医药卫生—病原生物学]