甲磺酸莫非赛定抑制HepAD38细胞内乙型肝炎病毒共价闭合环状DNA转录活性的研究  

The inhibition effect of GLS4JHS on the transcription activity of covalently closed circular DNA in HepAD38 cells

作　　者：董晓琴[1] 张英俊 任青云 李静 李勇 陈允甫 王贵强[1] 赵鸿[1] 

机构地区：[1]北京大学第一医院感染科,100034 [2]东阳光药业有限公司药业研究院

出　　处：《中华传染病杂志》2017年第5期290-293,共4页Chinese Journal of Infectious Diseases

摘　　要：目的初步探索HBV核心蛋白抑制剂甲磺酸莫非赛定（GLS4JHS）抑制HBV复制的机制。方法以HepAD38细胞为研究模型，用荧光定量PCR法检测不同浓度GLS4JHS，作用下细胞内前基因组RNA（pgRNA）和共价闭合环状DNA（cccDNA）水平的变化；用染色质免疫共沉淀法检测cccDNA微染色体上招募的核心蛋白及宿主组蛋白的变化。结果随着GLS4JHS浓度的增加，cccDNA和pgRNA的量依次减少；当药物浓度为400 nmol/L时，两者分别下降了94%和84%。同时，GLS4JHS可以降低结合于cccDNA微染色体的核心蛋白量以及乙酰化组蛋白量。结论GLS4JHS通过抑制cccDNA微染色体与HBV核心蛋白以及乙酰化组蛋白H3结合，降低cccDNA的转录活性，减少转录生成的pgRNA，从而降低HBV DNA的合成。Objective The hepatitis B virus （HBV） core protein assembly inhibitors GLS4JHS could destroy HBV eapsid assembly and the formation of non-capsid polymer structure. The aim of this study is to explore the mechanisms of GLS4JHS in inhibiting HBV replication. Methods HepAD38 cells was used as the study model. TaqMan real-time polymerase chain reaction （PCR） and quantitative real time PCR with specific primers were used to measure the change in pregenomie RNA （pgRNA） and covalently closed circular DNA （eecDNA） levels under different concentrations. ChIP assay in HepAD38 cells was used to assess the recruitment of HBV core protein and histone modifications. Results The amount of cccDNA and pgRNA decreased with the increasing GLS4JHS concentrations. After the drug concentrations reached 400 nmol/L, cccDNA and pgRNA declined by 94% and 84%respectively. Both HBV core protein occupancy on the cccDNA and cccDNA-bound H3 histone acetylation were reduced by GLS4JHS. Conclusions GLS4JHS decreases transcriptional activity of cccDNA and reduces pgRNA production by inhibiting cccDNA minicbromosome bound to HBV core protein and acetylated histone H3, which results in HBV DNA formation.

关 键 词：肝炎病毒 乙型 核心蛋白抑制剂 甲磺酸莫非赛定 共价闭合环状DNA 

分 类 号：R373.21[医药卫生—病原生物学]