Hayashi N, Takehara T, Kamada T
First Department of Medicine, Osaka University School of Medicine, Japan.
Princess Takamatsu Symp. 1995;25:143-9.
The lack of a small animal model for hepatitis C virus (HCV) infection has impeded elucidation of the pathogenesis of this virus. The aim of this study was to develop an HCV-expressing animal model using cationic liposome-mediated in vivo gene transfer. To examine the feasibility of this strategy, an expression vector composed of the LacZ gene driven by the beta-actin promoter, pActLacZ, was injected retrogradely into the common bile ducts of adult rats. X-Gal histochemical staining clearly showed that the LacZ gene was expressed in hepatocytes. Maximal expression was observed at a DNA:lipofectin ratio of 1:4. Based on this observation, an expression vector containing the full-length of HCV cDNA, pAGS3M091, was evaluated in adult rats. Two days after intrabiliary administration of pAGS3M091, PCR amplification of reverse-transcribed liver RNA demonstrated the 5' and 3' portions of HCV transcripts derived from pAGS3M091. Immunohistochemical analysis revealed the HCV core protein in a small number of hepatocytes scattered in the lobules. Thus, the full-length of the HCV genome was successfully expressed in adult rat liver using liposome-mediated in vivo gene transfer.
丙型肝炎病毒（HCV）感染缺乏小动物模型阻碍了对该病毒发病机制的阐明。本研究的目的是利用阳离子脂质体介导的体内基因转移建立一种表达HCV的动物模型。为了检验该策略的可行性，将由β-肌动蛋白启动子驱动的LacZ基因组成的表达载体pActLacZ逆行注入成年大鼠的胆总管。X-Gal组织化学染色清楚地表明LacZ基因在肝细胞中表达。在DNA:脂质体比例为1:4时观察到最大表达。基于这一观察结果，在成年大鼠中评估了含有HCV cDNA全长的表达载体pAGS3M091。在胆管内给予pAGS3M091两天后，对逆转录的肝脏RNA进行PCR扩增，证实了源自pAGS3M091的HCV转录本的5'和3'部分。免疫组织化学分析显示在小叶中散在的少数肝细胞中有HCV核心蛋白。因此，利用脂质体介导的体内基因转移成功地在成年大鼠肝脏中表达了HCV基因组的全长。
