摘要
目的 :探讨硫修饰和脂质体包裹对反义寡聚核苷酸 (antisenseoligodeoxynucleotides,AODN)抑制单纯疱疹病毒Ⅰ型 (herpessimplexvirusⅠ ,HSV1)复制的影响。方法 :将AODN、反义寡聚核苷酸脂质体 (antisenseoligodeoxynucleotideslipo some ,AODNL)、硫代反义寡聚核苷酸 (antisensephosphorothioatoligodeoxynucleotides,SAODN)和硫代反义寡聚核苷酸脂质体(antisensephosphorothioatoligodeoxynucleotidesliposome ,SAODNL)配制成 2 0、10、5、2 5和 1 2 5 μg/ml,加入至 96孔培养板 ,再加入 10 2 组织培养半数感染量 (5 0 %tissuecultureinfectivedosage,TCID50 )病毒 ,培养 2 4小时 ,细胞冻融后取上清液 ,加至 2 4孔培养板的细胞中 ,吸附 1小时 ,去病毒液 ,加入空斑上层液 ,培养 72小时 ,空斑计数。做病毒阳性对照 ,计算药物的病毒抑制率。结果 :各组药物浓度与抗病毒药效之间无相关性 (P >0 0 5 )。AODN组与AODNL组、SAODN组和SAODNL组相比有显著性差异 (P <0 0 5 ) ,SAODN组与SAODNL组之间有显著性差异 (P <0 0 5 ) ,AODNL组与SAODNL组间无显著性差异 (P>0 0 5 )。结论 :脂质体包裹、硫修饰或两者结合抑制病毒复制的作用都显著强于游离反义寡聚核苷酸 。
Objective:To evaluate effects on the antisense oligodeoxynucleotides inhibiting herpes simplex virus Ⅰ(HSV1) replication by the phosphorothioate modify and the liposome encapsulation.Methods:10 2TCID 50 (50% tissue culture infective dosage) virus and 20,10,5,2 5,1 25 μg/ml antisense oligodeoxynucleotides (AODN),antisense oligodeoxynucleotides liposome (AODNL),phosphorothioat antisense oligodeoxynucleotides (SAODN),phosphorothioat antisense oligodeoxynucleotides liposome (SAODNL) were added into 96 well cell culture plates.After the cells were cultured for 24 hours and frozen,the upper liquid was taken into 24 well cell culture plates.1 hour later,the virus was taken out and plaque liquid was added.The cells were culture for 72 hours and plaque was counted.The virus contrast was done and inhibitive rates were calculated.Results:The drug concentrations had no correlation to the effects.There were significant differences in AODN with AODNL,SAODN,and SAODNL.There were significant differences between SAODN and SAODNL,but no differences between AODNL and SAODNL.Conclusion:The inhibiting virus effects of phosphorothioate modify,liposome encapsulation and combination of both were stronger than those of free AODN.The inhibitive effects of liposome encapsulation were stronger than those of phosphorothioate modify.
出处
《眼科》
CAS
2002年第3期173-175,共3页
Ophthalmology in China
