摘要
探讨三螺旋形成寡核苷酸抗乙型肝炎病毒（ＨＢＶ）作用。方法针对ＨＢＶ核心启动子ＳＰＩ位点，台成２１ｍｅｒ硫代磷酸三螺旋形成寡核苷酸（ＴＦＯ２１）及２１ｍｅｒ无关对照寡核苦酸（ＯＤＮｃｏｎ）。采用ＥＬＩＳＡ、斑点杂交法分别检测了经寡核苷酸处理的ＨｅｎＧ２．２１５细胞（简称２．２１５细胞）及空白对照组细胞培养上清ＨＢｓＡｇ、ＨＢｅＡｇ及ＨＢＶＤＮＡ水平。结果ＴＦＯ２１组２．２．１５细胞ＨＢｓＡｇ、ＨＢｅＡｇ及ＨＢＶＤＮＡ分泌量明显低于空白对照组。ＴＦＯ２１对ＨＢｓＡｇ、ＨＢｅＡｇ的抑制分别达５７．５％、７７％。该抑制呈剂量依赖性，并与作用时间有关。而ＯＤＮｃｏｎ对ＨＢｓＡｇ、ＨＢｅＡｇ及ＨＢＶＤＮＡ均无影响、其对ＨＢｓＡｇ、ＨＢｅＡｇ的抑制率仅６％～９％，６％～８％。在实验范围内，硫代磷酸寡核酸对２２１５细胞无毒性作用。结论ＴＦＯ２１在体外能有效抑制ＨＢＶ复制及抗原合成，具有较大应用潜力。
Objective To explore the inhibitory effect of triplex forming oligodeoxynucleotides (T F O )on replication of HBV and synthesis of antigen. Methods A 21 mer phosphorothioate triplex formingoligodeoxynucleotides (TFO21) directed at SPl sites in HBV core promoter and a control of 21 inerphosphorothioate oligodeoxynucleotides (ODNcon) were synthesized. HepG 2.2.15 cells which can produceHBsAg, HBeAg, HBV DNA and HBV particle were treated with TFO21 and ODNcon. In HepG 2.2.15 cellstreated with these oligodeoxynucleotides, the levels of HBsAg, HBeAg, and HBV DNA were examined byELISA and dot hybridization method, respeCtively. Results The levels of HBsAg, HBeAg and HBV DNAin TFO21 gToup were lower than those in the bank control group. At concentration of 10 mol/L, TFO21inhibited snythesis of HBsAg and HBeAg by 57.5% and 77%. The inhibitory effect of TFO21 was dosagrdependent, and was related to time in which 2.2.15 cells were incubated with TFO21. No inhibition wasobserved in the ODNcon group. No toxicity was observed in the 2.2.15 cells treated witholigodeoxynucleotides. Conclusion These results indicate that TFO is a potent inhibitor for HBVreplication and synthesis of antigen, and also suggest that TFO is a therapeutic potential for the treatmentof patients infected with HBV.
出处
《中华肝脏病杂志》
CAS
CSCD
1999年第1期8-10,共3页
Chinese Journal of Hepatology
基金
广东省博士后基金
广东省科委重点科技攻关项目
国家教委博士点基金
关键词
乙型肝炎病毒
基因疗法
寡脱氧核苷酸
TFO
Hepatitis
B virus
Gene
therapy
Triplex
forming
oligodeoxynucleotides
(TFO )
