文章摘要
席力森,邓 蕾,张 勇,高 云,王学浩,孙倍成.慢病毒载体携带siRNAs对乙型肝炎病毒抑制作用的体内研究[J].南京医科大学学报,2009,29(3):317~322
慢病毒载体携带siRNAs对乙型肝炎病毒抑制作用的体内研究
Studies on inhibition of lentiviral vector carried siRNAs interfering against hepatitis B virus In vivo
投稿时间:2008-09-13  
DOI:10.7655
中文关键词: 小干扰RNA  慢病毒载体  肝炎病毒  乙型
英文关键词: siRNAs  lentiviral vectors  hepatitis B virus
基金项目:江苏省六大人才高峰课题(06-B-032);江苏省医学重点人才基金(RC2007057)
作者单位
席力森 南京医科大学第一附属医院肝脏外科,江苏 南京 210029 
邓 蕾  
张 勇  
高 云  
王学浩  
孙倍成  
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中文摘要:
      目的:建立小鼠急性乙型肝炎病毒感染动物模型,在此基础上观察慢病毒载体携带小干扰RNA(small interfering RNAs,siRNAs)对乙型肝炎病毒(HBV)复制和表达的影响?方法:36只Balb/c小鼠随机等分为3组(n = 12):①空白对照组(NC组):通过尾静脉注射pTHBV2(包含HBV全基因组真核表达质粒),建立小鼠急性HBV感染模型;②干扰组(E组):将pTHBV2与pWPT-HBV-siRNA1(携带靶向HBV S区的siRNA1的慢病毒)通过尾静脉共注射小鼠;③无关干扰组(I组):pTHBV2和pWPT-HBV-siRNA2(携带非靶向HBV的无关siRNA2的慢病毒) 通过尾静脉共注射小鼠?于注射后第4天和第7天取小鼠血清和肝脏组织?采用ELISA法检测血清中HBsAg(HBV表面抗原);选择逆转录聚合酶链反应方法(RT-PCR)检测肝组织HBV S-mRNA水平;用免疫组织化学法检测肝组织HBcAg(HBV核心抗原)和HBsAg?结果:与NC组比较,E组小鼠血清中HBsAg水平在第4和第7天分别下降89.76%和94.81%(P < 0.01);RT-PCR结果提示E组肝组织HBV S-mRNA水平明显降低(P < 0.05);免疫组织化学法检测结果显示E组肝组织HBcAg﹑HBsAg阳性细胞数明显减少(P < 0.05)?I组与NC组比较,上述检测指标无统计学差异(P > 0.05)?结论:慢病毒载体携带siRNAs能够显著抑制小鼠体内HBV的复制和表达,并且抑制作用在第7天较第4天更强,没有减弱的趋势?
英文摘要:
      Objective:To develop a mouse model of acute hepatitis B virus infection and observe lentiviral vectors(LVs) with small interfering RNAs-mediated inhibition of HBV replication and expression in the model. Methods:Thirty-six Balb/c mice were randomly divided into three groups(n = 12,each). ①Negative control group(NC group):the model of acute hepatitis B virus infection were established by administrating pTHBV2(the eukaryotic expression vectors which contain HBV genome informations) through tail-vein; ②experimental group(E group): pTHBV2 and pWPT-HBV-siRNA1(the lentivirus with siRNA1 aimed directly at HBV S-sector) were administrated through tail-vein; ③independence group(I group): pTHBV2 and pWPT- HBV-siRNA2(the lentivirus with siRNA2 which didn’t aim at HBV) were administrated by the same method. The serum and liver tissue were collected on the fourth and seventh days respectively. ELISA method was used to detect HBV surface antigen(HBsAg) in serum; RT-PCR was selected to detect HBV S-mRNA level in hepatic tissue. Intracellular HBV core antigen(HBcAg) and HBsAg were determined by immunohistochemistry. Results:In contrast with NC group, serum HBsAg level in E group was decreased by 89.76% and 94.81% on the fourth and the seventh days, respectively(P < 0.01);Liver tissue HBV S-mRNA level, intracellular HBcAg and HBsAg levels were also depressed obviously(P < 0.05); whereas serum HBsAg, tissue HBV S-mRNA,intracellular HBcAg and HBsAg levels didn’t differ significantly between NC group and I group(P > 0.05). Conclusion:Based on the results of this experiment, LVs with siRNAs could remarkably inhibit the replication and expression of HBV in vivo, which augmented from the fourth day to the seventh day.
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