乙型肝炎病毒转基因小鼠体内肝靶向反义RNA抗病毒疗效研究  

Anti-HBV effect of antisense RNA targeted to liver in HBV transgenic mice:

作　　者：陈枫[1] 郑素军[1] 钟森[1] 邓存良[1] 王明勇[1] 张建军[1] 廖国伟[1] 

机构地区：[1]泸州医学院附属医院,四川泸州646000

出　　处：《四川医学》2002年第8期833-835,共3页Sichuan Medical Journal

基　　金：四川省科委重点项目基金 [1 996] 0 2 6;四川省教委基金 [1 996] 0 2

摘　　要：目的　探讨半乳糖修饰的反义 RNA真核表达质粒在乙型肝炎病毒转基因小鼠体内的抗病毒作用。方法　以半乳糖多聚赖氨酸 (Gal- PL L )作肝靶向载体 ,将乙型肝炎病毒基因 C区的反义 RNA真核表达重组质粒 (p CEP4 -a C)制备为 Gal- PL L - p CEP4 - a C。将 2 4只血清 HBV DNA、HBs Ag阳性的小鼠 ,随机等分为 Gal- PL L - p CEP4 - a C治疗组、Gal- PL L - p CEP4对照组和生理盐水阴性对照组 ,于实验第 1天尾静脉分别注射 Gal- PL L - p CEP4 - a C、Gal- PL L - p CEP4(10 0μg/只 )和等体积的生理盐水 ,观察治疗前后血清 HBV DNA以及 HBs Ag变化。结果　 Gal- PL L - p CEP4 - a C治疗组2 1天时血清 HBV- DNA转阴率 6 2 .5 % (5 / 8) ,且 7,14 ,2 1天时血清 HBs Ag明显降低 ;而 Gal- PL L - p CEP4组血清 HBVDNA转阴 1只 (1/ 8) ,生理盐水组 8只均未转阴 ,两组用药后血清中 HBs Ag与用药前比较差异性均不明显 (P>0 .0 5 )。结论　肝靶向反义 RNA能在乙肝基因小鼠体内抑制Objective To investigate the anti HBV effect of targeted antisense RNA in HBV transgenic mice.Methods pCEP4 aC which could transcript antisense RNA against HBV C gene in eukaryotic cells was incubated with glactosylated poly L lisine(Gal PLL)in the ratio of 1μg to 0.8μg for 0.5 hour to produce the targeted Gal PLL pCEP4 aC.Twenty four mice whose HBV DNA,HBsAg were positive were randomly divided into 3 groups,8 in each group.The treatment group and the posteive control group were injected 100μg of Gal PLL pCEP4 aC and 100μg of Gal PLL pCEP4 through tail vein respectively for per mice at the 1st day;while in the negative control group,each mice received the same volume of normal saline by the same way.HBV DNA,HBsAg in serum of mice at pretreatment and posttreatment were detected with ELISA and Nested PCR respectively.Results At the 21st day,the serum HBV DNA become negative in 5/8 of the pCEP4 aC treated mice,but only in 1/8 of pCEP4 treated mice and in 0/8 of the negative control group.In treatment group,the serum HBsAg were decreased,significant difference was found at the 7th day ( P <0.05),at the 14th day,and 21st day ( P <0.01)compared with those of pretreatment,while serum HBsAg in control groups had no apparent changes ( P >0.05).Conclusion The antisense RNA targeted to the liver can inhibit the replication of HBV DNA and decrease the content of HBsAg,HBcAg.

关 键 词：乙型肝炎病毒 反义RNA 靶向治疗 半乳糖多聚赖氨酯 乙型肝炎 基因疗法 动物实验 

分 类 号：R512.62[医药卫生—内科学]