雄黄纳米微粒在细胞水平上抑制呼吸道合胞病毒复制的初步研究  

Study on the Effect of Realgar Nanoparticles on Reducing the Respiratory Syncytial Virus Type A(RSV-A) Replication in Vitro

作　　者：程淼[1,2] 赵洪兰[2] 王成祥[1] 王惠芳[3] 张燕[2] 苟宝迪[4] 朱贞[2] 王明哲[1] 许文波[2] 

机构地区：[1]北京中医药大学东直门医院,北京100700 [2]中国疾病预防控制中心病毒病预防控制所,北京102206 [3]解放军防化研究院第四所,北京102205 [4]北京大学药学院化学生物学系,北京100083

出　　处：《病毒学报》2012年第1期45-50,共6页Chinese Journal of Virology

基　　金：病毒基因工程国家重点实验室开放课题"雄黄在细胞水平上对呼吸道合胞病毒和腺病毒作用的评价"

摘　　要：建立呼吸道合胞病毒A型(RSV-A型)感染Hep-2细胞模型,通过预防、治疗及直接灭活三种不同给药方式,观察中药雄黄对RSV-A型感染Hep-2细胞病变(CPE)的抑制作用。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RSV-A型感染Hep-2细胞病变所起的作用,并对药物的量效关系进行分析。雄黄纳米微粒TC50值为0.649μg/mL。预防、治疗及直接灭活给药方式均可减轻RSV-A感染Hep-2细胞的CPE程度,其抑制RSV-A型感染Hep-2细胞病变的半数有效浓度(IC50)分别为0.20μg/mL、0.13μg/mL、0.16μg/mL,治疗指数(TI)分别为3.18、4.99和4.11,雄黄纳米微粒对RSV-A型感染Hep-2细胞病变的抑制作用存在着明显的量效关系。雄黄纳米微粒按预防、治疗及直接灭活给药方式给药时,其中治疗给药方式更有利于减轻RSV-A感染Hep-2细胞引起的病变。This study was to establish a model to explore anti-RSV effect of different administration method of Chinese medicine realgar on respiratory syncytial virus type A（RSV-A） replication in Hep-2 cells.Using high-energy ball milling with distilled water to prepare realgar nanoparticles,the concentration of nanometer realgar was tested by molybdenum blue staining method and the size of realgar nanoparticles was tested on Nano Series.Cell culture with ribavirin as a positive control was applied to observe the effect of anti-respiratory syncytial virus type A replication through prevention,treatment or direct inactivation of three different drug administration methods.Realgar nano-particles was found to be a potential inhibitor of RSV-A in a concentration-dependent manner with the median toxic concentration（TC50） of 0.649μg/mL in Hep-2 cell culture.The median inhibition concentration（IC50） was 0.20μg/mL when drug was added before virus infection.The IC50 was 0.13μg/mL when drug was added after virus infection,and it was 0.16μg/mL when the drug was mixed with virus and added.The therapeutic index（TI）was 3.18,4.99 and 4.11,respectively.The results showed realgar nanoparticles could inhibit the replication of the RSV and inactivate the RSV in vitro.

关 键 词：呼吸道合胞病毒 雄黄纳米微粒 细胞病变效应 量效-关系 

分 类 号：R373.1[医药卫生—病原生物学]