纳米银体外抗腺病毒作用初步研究

目的探讨纳米银(silver nanoparticles,silver-nps)在体外对腺病毒3型(Adenovirus type 3,ADV3)的抑制作用。方法运用细胞培养技术、MTT测值法、CPE和免疫荧光观察法,分析纳米银对ADV3感染HeLa细胞的预防作用、直接灭活作用以及对ADV3子代病毒体生成的抑制作用。结果纳米银能明显杀伤ADV3,且呈剂量依赖性;纳米银在HeLa细胞上最大无毒浓度(TC0)为52.48μg/ml;ADV3在HeLa细胞上的组织半数感染量(TCID50)为10-2.74/100μl;最大无毒剂量范围内,50、25、12.5、6.25和3.125μg/ml的纳米银分别与100 TCID50ADV3等体积混合,细胞存活率分别为(95.38±2.60)%、(60.51±9.42)%、(57.99±8.72)%、(41.35±3.91)%和(37.88±3.75)%,而100 TCID50ADV3感染HeLa细胞后,测得细胞存活率为(31.92±8.98)%,二者相比差异有统计学意义(P0.05);免疫荧光结果显示,与纯病毒组形成的强特异性荧光相比,纳米银在ADV3吸附细胞后加入、吸附前预处理HeLa细胞和纳米银同ADV3同时作用三种途径特异性荧光都很少见,说明纳米银在不同途径下对ADV3均有抑制作用。结论纳米银对腺病毒3型具有明显的抑制作用。

A preliminary study of anti-adenovirus effect of silver nanoparticles in vitro

Objective To evaluate the inhibiting effect of silver nanoparticles（silver-nps） against adenovirus type3（ADV3）.Method The anti-adenovirus activities of silver-nps in HeLa cells was determined by means of MTT assay,cytopathic effect and immunofluorescence test,which was used to analyze the anti-adenovirus activities of silver-nps in preventing and inactivating HeLa cells infected with ADV3 and inhibiting the formation of ADV3 progeny virions.Result The silver-nps showed effective inhibiting activity to ADV3 in a concentration-dependent manner.TC0 of silver-nps was 52.48 μg/ml.The median tissue culture infective dose（TCID50） of ADV3 on HeLa cell was 10-2.74.In the range of non-toxic concentration,the survival rates of HeLa cells were（95.38±2.60）%,（60.51±9.42）%,（57.99±8.72）%,（41.35±3.91）% and（37.88±3.75）% after silver-nps at different concentrations of 50,25,12.5,6.25 and 3.125 μg/ml were mixed with 100 TCID50 of ADV3 in 2 hours,while the survival rate of control group treated with 100 TCID50 of ADV3 was（31.92±8.98）%（P〈0.05）;HeLa cells infected with ADV3 had much specific immunofluorescence when compared to the cells treated with silver-nps in different ways,which indicated that the silver-nps had inhibiting effect on ADV3.Conclusion Silver-nps has remarkably inhibiting effects on adenovirus type 3.