中文摘要：

目的：观察 HBV-三磷酸（3p）-小干扰 RNA（siRNA）激活受试小鼠Ⅰ型干扰素抗病毒天然免疫反应以及抑制 HBV 基因组复制的双重作用。方法针对 HBV 基因组 S/P mRNA 设计并通过体外转录法制备带有5′-3p-修饰的 HBV-3p-siRNA；以未经修饰的 HBV-siRNA 和阴性对照（NC）-siRNA 作为对照；利用尾静脉水动力注射法建立 HBV 感染小鼠模型。将40只小鼠分为4组，每组10只，模型组：仅注射 pGL3．0-HBV 1．2拷贝质粒；阴性对照组：腹腔注射 NC-siRNA；HBV-siRNA 组：腹腔注射 HBV-siRNA；HBV-3p-siRNA 组：腹腔注射 HBV-3p-siRNA。ELISA 法检测小鼠血清中HBsAg 的表达和β干扰素的分泌量；荧光定量 PCR 检测小鼠血清 HBV DNA。采用 LSD 或 Dunnett T3统计方法进行 One way-ANOVA 统计学分析。结果小鼠血清中β干扰素水平模型组为（12．37±5．32）pg/mL，阴性对照组为（22．61±6．29）pg/mL，HBV-siRNA 组为（26．40±5．39）pg/mL，HBV-3p-siRNA 组为（68．37±21．00）pg/mL，HBV-siRNA 组和 HBV-3p-siRNA 组与模型组相比，差异有统计学意义（F 值分别为23．988、46．523，均 P ＜0．01）。小鼠血清中 HBsAg 水平模型组为（2864．86±907．11）ng/mL，阴性对照组为（2198．86±456．89）ng/mL，HBV-siRNA 组为（1049．71±396．28）ng/mL， HBV-3p-siRNA 组为（640．86±383．08）ng/mL，HBV-siRNA 组和 HBV-3p-siRNA 组与模型组相比，差异有统计学意义（F 值分别为23．537、39．144，P 值分别为0．025、0．010）。小鼠血清中 HBV DNA 的水平模型组为（2．54×104^±1．46×10^4）拷贝/mL，阴性对照组为（2．22×10^4±2．62×10^3）拷贝/mL， HBV-siRNA组为（3．59×10^3±2．88×10^3）拷贝/mL，HBV-3p-siRNA 组为（2．65×10^3±1.46×10^3）拷贝/mL，HBV-siRNA 组和 HBV-3p-siRNA 组与模型组相比，差异均有统计学意义（F 值分别为15．013、16．741，均 P ＜0．05）。HBV-3p-siRNA、HBV-siRNA，以及 NC-siRNA 对 HBV 感

英文摘要：

Objective To observe the activation of anti-viral innate immune response of type Ⅰinterferon and inhibition of hepatitis B virus （HBV）genome replication in mice by HBV-3p-siRNA. Methods HBV-3p-siRNA was designed by targeting specific sequence of HBV S/P mRNA and was generated by in vitro transcription.Negative control siRNA （NC-siRNA）and non-modified HBV-siRNA were used as control groups.Bloosamples were collected from tail vein of mice and the model of HBV-infected mice were established by hydrodynamic injection.Forty mice were divided into 4 groups with 10 in each group.The model group was only injected with pGL3.0-HBV1 .2 copy plasmid.The negative control group received peritoneal injection of NC-siRNA.HBV-siRNA group received peritoneal injection of HBV-siRNA and HBV-3p-siRNA group received peritoneal injection of HBV-3p-siRNA.The interferon-β（IFN-β）and hepatitis B surface antigen （HBsAg）in serum were detected by enzyme linked immunosorbent assay （ELISA）.The copies of HBV DNA were assessed by fluore scence quantitative polymerase chain reaction （PCR ）.The statistical difference between groups was determined using One way-ANOVA analysis by LSD or Dunnett T3.Results Serum level of IFN-β was （12.37±5 .32）pg/mL in model group,（22.61 ±6.29 ）pg/mL in negative control group,（26.40±5 .39）pg/mL in HBV-siRNA group and （68.37± 21 .00 ） pg/mL in HBV-3p-siRNA group.The secretions of IFN-β into serum were significantly enhanced by HBV-siRNA and HBV-3p-siRNA compared with model group （F =23.988 and 46.523,respectively,both P 〈0.01）.Serum level of HBsAg was （2 864.86±907.11 ）ng/mL in model group,（2 198.86±456.89 ）ng/mL in negative control group,（1 049.71 ± 396.28 ）ng/mL in HBV-siRNA group and （640.86±383.08）ng/mL in HBV-3p-siRNA group.The expressions of HBsAg were inhibited by HBV-3p-siRNA and HBV-siRNA compared with model group （F = 23.537 and 39.144, respectively;P =0.025 and 0.010,respectively）.Serum level of HBV DNA was （2.54 ×10^4 ±1 .46