杨林 陈幼明 高志良 邓练贤 陈雪娟 姚集鲁 510630,广州,中山医科大学附属第三医院病毒性肝炎研究室 中华肝脏病杂志 1999 0 7 1


    关键词：乙型肝炎病毒 基因疗法 三螺旋形成寡脱氧核苷酸(TFO) 期刊 zhgzbzz 0 病毒性肝炎新治疗 fur -->


    

摘要 目的 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SP1位点，合成21 mer硫代磷酸三螺旋形成寡核苷酸(TFO₂₁ )及21 mer无关对照寡核苷酸(ODNcon)。采用ELISA、斑点杂交法分别检测了经寡核苷酸处理的HepG2.2.15细胞(简称2.2.15细胞)及空白对照组细胞培养上清HBsAg、HBeAg及HBV DNA水平。结果 TFO₂₁ 组2.2.15细胞HBsAg、HBeAg及HBV DNA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HBV DNA均无影响，其对HBsAg、HBeAg的抑制率仅6%～9%，6%～8%。在实验范围内，硫代磷酸寡核苷酸对2.2.15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。

    

Inhibitory effect of triplex forming oligodeoxynucleotides on HBV replication and synthesis of antigen

YANG Lin,CHEN Youming,GAO Zhiliang,et al.

Viral Hepatitis Research Unit,The Third Affiliated Hospital.Sun Yat-sen University of Medical Sciences,Guangzhou 510630

Abstract Objective To explore the inhibitory effect of triplex forming oligodeoxynucleotides(TFO) on replication of HBV and synthesis of antigen.Methods A 21 mer phosphorothioate triplex forming oligodeoxynucleotides (TFO₂₁ ) directed at SP1 sites in HBV core promoter and a control of 21 mer phosphorothioate oligodeoxynucleotides (ODNcon) were synthesized.HepG 2.2.15 cells which can produce HBsAg,HBeAg,HBV DNA and HBV particle were treated with TFO₂₁ and ODNcon.In HepG 2.2.15 cells treated with these oligodeoxynucleotides,the levels of HBsAg,HBeAg,and HBV DNA were examined by ELISA and dot hybridization method,respectively.Results The levels of HBsAg,HBeAg and HBV DNA in TFO₂₁ group were lower than those in the bank control group.At concentration of 10 mmol/L,TFO₂₁ inhibited snythesis of HBsAg and HBeAg by 57.5% and 77%.The inhibitory effect of TFO₂₁ was dosage-dependent,and was related to time in which 2.2.15 cells were incubated with TFO₂₁ .No inhibition was observed in the ODNcon group.No toxicity was observed in the 2.2.15 cells treated with oligodeoxynucleotides.Conclusion These results indicate that TFO is a potent inhibitor for HBV replication and synthesis of antigen,and also suggest that TFO is a therapeutic potential for the treatment of patients infected with HBV.

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