针对HBV preS1、preS2编码链的反基因锁核酸对HepG2.2.15细胞的抗病毒效果  

Antiviral effect of antigene locked nucleic acid of HBV preS1,preS2 coding chain on Hep2.2.15 cells

作　　者：彭彬 许桂丹[1] 韦武均 农顺强 陈晓昊 肖树荣 邓益斌 PENG Bin;XU Guidan;WEI Wujun;NONG Shunqiang;CHEN Xiaohao;XIAO Shurong;DENG Yibin(Department of Clinical Laboratory,Affiliated Hospital of Youjiang Medical College for Nationalities,Baise,Guangxi 533000,China;Guangxi Hepatobiliary Disease Clinical Medical Research Center,Baise,Guangxi 533000,China)

机构地区：[1]右江民族医学院附属医院检验科,广西百色533000 [2]广西肝胆疾病临床医学研究中心,广西白色533000

出　　处：《检验医学与临床》2020年第21期3106-3109,共4页Laboratory Medicine and Clinic

基　　金：国家自然科学基金项目(81460123);广西科技计划项目基地与人才专项(桂科AD17129025);广西自然科学基金项目(2018GXNSFAA281187);广西肝胆疾病临床医学研究中心(桂科AD17129025-25)。

摘　　要：目的探讨乙型肝炎病毒(HBV)preS1、preS2 DNA同聚嘌呤区设计的锁核酸在体外对HBV复制和表达的抑制作用。方法设计并合成锁核酸,以Lipo3000作为载体转染至锁核酸-脂质体组,设空白对照组,运用荧光定量PCR、化学发光免疫分析等技术观察锁核酸对HBsAg表达、HBV DNA复制的抑制作用,采用CCK8比色法观察锁核酸对HepG2.2.15细胞基本代谢的影响。结果加入锁核酸后,HBsAg、HBV DNA表达量相对于空白对照组逐渐降低,并随时间呈递增趋势,其中,preS1的3023-3037nt位点在第9天对HBsAg表达的抑制率达(61.94±4.11)%,对HBV DNA复制的抑制率达(56.08±3.22)%;preS2的3305-3320nt位点在第9天对HBsAg表达的抑制率达(72.93±4.14)%,对HBV-DNA复制的抑制率达(61.79±3.40)%。结论HBV preS1编码链的3023-3037nt位点和preS2编码链的3305-3320nt位点的反基因锁核酸片段对HBsAg表达和HBV-DNA复制的抑制效果最为明显,两者均可以作为抗HBV治疗的有效靶位。Objective To investigate the inhibition of HBV replication and expression in vitro by the oligopurine region of preS1 and pres2 DNA of hepatitis B virus(HBV).Methods Designed and synthesized locked nucleic acid,transfered to locked nucleic acid-liposome group with Lipo3000 as carrier,set up a blank control control,used fluorescent quantitative PCR,chemical luminescent immunoassay and other technologies to observe locked nucleic acid′s inhibition of HBsAg expression,HBV DNA replication,Finally,the effect of locked nucleic acid on the basic metabolism of HepG2.2.15 cells was observed by CCK8 color method.Results After adding locked nucleic acid,the expression of HBsAg and HBV DNA gradually decreased compared with the control group,and showed an increasing trend with time.Among them,the 3023-3037nt site of preS1 inhibited HBsAg expression by(61.94±4.11)%on the 9th day,the inhibition rate of HBV DNA replication reached(56.08±3.22)%;the 3305-3320nt sites of preS2 inhibited HBsAg expression by(72.93±4.14)%on the 9th day,the inhibition rate of HBV-DNA replication reached(61.79±3.40)%.Conclusion Antigen-locked nucleic acid fragments targeting 3023-3037nt sites of HBV preS1 coding chain and 3305-3320nt sites of preS2 coding chain have the most obvious inhibitory effects on HBsAg expression and HBV-DNA replication,and both of which can be used as effective targets for anti-HBV therapy.

关 键 词：乙型肝炎病毒 锁核酸 PRES1 PRES2 HEPG2.2.15细胞 

分 类 号：R446.9[医药卫生—诊断学]