硫代反义寡核苷酸在鸭体内外对鸭乙型肝炎病毒DNA复制的影响  

Inhibition of duck hepatitis B virus DNA replication by antisense phosphorothioate oligodeoxynucleotides in vitro and in vivo

作　　者：董飚[1] 邵兴无[1] 陶佩珍[1] 

机构地区：[1]中国医学科学院中国协和医科大学医药生物技术研究所,北京100050

出　　处：《中华实验和临床病毒学杂志》2003年第1期25-27,共3页Chinese Journal of Experimental and Clinical Virology

基　　金：中国医学科学院青年重点基金资助 (9332 0 0 4)

摘　　要：目的　研究在转录起始区域设计硫代反义寡核苷酸抑制鸭乙型肝炎病毒DNA(DHBVDNA)复制的可行性。方法　设计合成分别基于DHBVPreS1、PreS2、S抗原启动基因的硫代反义寡核苷酸 ,应用Southernblot实验和cpm计数测定其对原代鸭肝细胞中及雏鸭中感染DHBV的DHBVDNA的作用。结果　在 1.5 μmol L浓度下 ,体外抑制率分别是 61.5 % ,69.3 %和 62 .4%。选取PreS1抗原基因起始区段的硫代反义寡核苷酸进行整体动物实验。每只动物每天按 10 μg g体重静脉注射一次 ,共给药 6d。对动物肝脏进行取样分析表明 ,在此剂量下 ,对DHBVDNA的抑制率可达 87.9%。结论　本实验所设计的针对转录起始区的硫代反义寡核苷酸在体内外对DHBVDNA复制均有明显抑制作用 。Objective To determine the feasibility of inhibition of duck hepatitis B virus (DHBV) DNA replication by antisense phosphorothioate oligodeoxynucleotides corresponding to DHBV transcription region. Methods The authors designed three antisense phosphorothioate oligodeoxynucleotides which correspond to DHBV PreS1,PreS2 and S antigen gene promotors respectively. The DNA replication level was detected with Southern blot method and cpm calculation. Results Primary duck hepatocyte culture was treated with 1.5 μmol/L antisense oligo deoxynucleotides in vitro , all the antisense fragments caused a firm inhibition of viral DNA replication and the inhibition rates were 61.5%,69.3% and 62.4%, respectively. In vivo ,the animals were treated with 10 μg/g PreS1 antigen gene promotor antisense oligodeoxynucleotides per day for 6 days and a very strong inhibition rate of 87.9% was obtained .Conclusion The results demonstrated the pontential ceinical application of antisense phosphorothioate oligodeoxynucleotides in clincs.

关 键 词：寡核苷酸类 乙型肝炎病毒 鸭 抗病毒药 

分 类 号：R346[医药卫生—基础医学]