反义寡聚硫代磷酸型、甲基磷酸型核苷酸体外抑制 HSV-Ⅰ增殖与抗原表达  

Suppression of herpes simplex virus type Ⅰ proliferation and antigen expression by antisense oligodeoxynucleotides of phosphorothioates and methylphosphonates in vitro

作　　者：余虹[1,2] 任中原[1,2] 宗建超 倪爱国[1,2] 黄慧玲 

机构地区：[1]天津医科大学微生物学教研室 [2]南开大学分子生物学研究所

出　　处：《中华微生物学和免疫学杂志》1997年第3期164-167,共4页Chinese Journal of Microbiology and Immunology

基　　金：国家自然科学基金

摘　　要：以人单纯疱疹病毒（ＨＳＶ）基因组即刻早期ｍＲＮＡ４和５拼接区１０３４～１０５０密码子序列为靶位点，人工化学修饰、合成特异性８～１４ｍｅｒ反义寡聚硫代磷酸型、甲基磷酸型及未修饰型核苷酸（Ｓ－ＯＤＮ１４、Ｍ－ＯＤＮ８、Ｎ－ＯＤＮ８）体外抑制病毒活性。Ｓ－ＯＤＮ１４５～１０μｍｏｌ／Ｌ浓度即可抑制ＨＳＶ致ＣＰＥ和ＰＦＵ，Ｍ－ＯＤＮ８４０～６０μｍｏｌ／Ｌ表现明显抑制效应，ＥＬＩＳＡ检测Ｓ－ＯＤＮ１４１０μｍｏｌ／Ｌ接近５０％抑制病毒抗原表达。Ｍ－ＯＤＮ８的抑制病毒活性剂量呈现细胞毒性。We synthesized modified antisense phosphorothioates and methylphosphonates oligodeoxynucleotides(S ODN 14mer,M ODN 8mer)which are complementary to the splice junction of herpes simplex virus type Ⅰ(HSV Ⅰ) immediate early pre mRNA 4 and 5 gene.The oligodeoxynuleotides were tested for their ability to inhibit cytopathogenic effect,viral antigen expression of HSV Ⅰ and plaque formation reduction in vitro.Antisense ODN was administered with different doses(5μmol/L,10μmol/L,15μmol/L,20μmol/L,40μmol/L,,60μmol/L,)in different times of prior or post infection.The results showed that 10μmol/L of S ODN 14 had the ability to inhibit the cytopathogenic effect caused by HSV Ⅰ.The 50% effective dose(ED 50 ) was 20μmol/L and ED 90 was 40μmol/L respectively.The concentration of S ODN 14 which reduced HSV Ⅰ induced plaque forming unit by 50% in Vero cells was 20μmol/L and inhibitory rate of 40μmol/L S ODN 14 was 89%,Detection of viral antigen expression by ELISA showed that 10μmol/L of S ODN 14 inhibitory rate by 48% and 20μmol/L￣40μmol/L by 58%￣75%.The protection of ODN for cell against HSV Ⅰ showed,in virus replicative cycle,stronger antiviral activity at early phase of the viral replication and weaker at the late phase.

关 键 词：寡核苷酸 反义核酸 单纯疱疹病毒 抑制 病毒 

分 类 号：R977[医药卫生—药品] R373.11[医药卫生—药学]