雄黄纳米微粒在细胞水平上抑制腺病毒复制的初步研究

Study on the effect of realgar nanoparticles anfi-adenovirus in vitro

目的 建立腺病毒3型（ HAdV-3）感染Hep-2细胞模型,观察中药雄黄对腺病毒3型（ HAdV-3）感染Hep-2细胞病变的抑制作用.方法 用高能球磨机研磨双蒸水水飞处理制备雄黄纳米微粒,应用砷钼蓝染色法测定雄黄纳米微粒浓度并在Nano Series粒度测定仪上测定其粒度.以MTT法计算药物的半数中毒剂量（TC50）.通过三种不同给药方式即预防给药、治疗给药及直接灭活给药方式进行体外实验,以利巴韦林为阳性对照药,观察雄黄纳米微粒对HAdV-3感染Hep-2细胞病变所起的作用,并对药物的量效关系进行分析.结果 雄黄纳米微粒TC50值为0.649 μg/ml.预防、治疗及直接灭活给药方式均可减轻HAdV-3感染Hep-2细胞的CPE程度,其抗HAdV-3的半数有效浓度（ IC50）分别为0.255 μg/ml、0.142 μg/ml、0.117 μg/ml,治疗指数（TI）分别为2.55、4.57和5.55,雄黄纳米微粒对HAdV-3感染Hep-2细胞CPE的抑制作用存在着明显的量效关系.结论 雄黄纳米微粒在体外有抑制HAdV-3病毒复制和直接灭活病毒的作用,并有一定保护Hep-2细胞预防HAdV-3病毒感染的作用.

Objective This study was to establish a model that adenovirus type 3 （HAdV-3） infected on Hep-2 cell in order to explore anti-adenovirus3 （ HAdV-3 ） effect of Chinese medicine realgar in vitro.Method Use high-energy ball milling with distilled water to prepare realgar nanoparticles.The concentration of nanometer realgar was tested by molybdenum blue staining method and realgar nanoparticles＇ particle size was tested on Nano Series.The techinique of cell culture with ribavirin as positive control was to observe anti-adenovirus effect through prevention,treatment and direct inactivation of three kinds of drug delivery.Result This drug was found to be a potential inhibitor of HAdV-3 in a concentration-dependent manner with the median toxic concentration （ TC50 ） of 0.649 μg/ml in Hep-2 Cell culture.The median inhibition concentration（ IC50 ） was 0.255 μg/ml when drug was added before infection.The IC50 was 0.142 μg/ml when drug was added after virus infection,and it was 0.117 μg/ml as the drug was added after it mixed with virus.The therapeutic index （ TI ） was 2.55,4.57and 5.55 respectively.Conclusion The direct inactivation effect of realgar nanoparticles is the most evident in three drug deliveries manner with the same concentration in vitro.