Pathogenesis of liver injury and carcinogenesis in HCV-expressing model animals by in vivo gene transfer.

通过体内基因转移表达 HCV 模型动物的肝损伤和癌变的发病机制。

• 批准号: 05404030
• 负责人: KAMADA Takenobu
• 金额: $ 14.59万
• 依托单位: Osaka University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (A)
• 财政年份: 1993
• 资助国家: 日本
• 起止时间: 1993 至 1995
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-05404030/
• 关键词: in vivo gene transfer; hepatitis C virus; cationic liposome; asialoglycoprotein; rat; liver injury; hepatocarcinogenesis; expression vector; 肝発泡機構; β-ガラクトシダーゼ; C形肝炎ウィルス; in vivo遺伝子導入; リポソーム; PCR; 免疫組織化学; Lac Z; 肝炎; 肝癌

The lack of small animal model of hepatitis C virus (HCV) infection impeded elucidation of the pathogenesis of this virus. The aim of this study is to develop HCV-expressing model animals using in vivo gene transfer and use them to study the pathogenesis of liver injury and hepatocarcinogenesis. In this study, we used cationic liposome-mediated and asialoglycoprotein receptor-mediatd in vivo gene transfer. To examine the feasibility of these procedures, pActLacZ in which the LacZ gene was driven by the beta-actin promoter was used as an reporter vector and successful transfection into rat hepatocytes in vivo was confermed by X-Gal histochemistry. For expression of HCV,HCV-expression vector in which the full-coding sequence of the HCV genome was driven by the beta-actin promoter was used. The expression of HCV in rat hepatocytes in vivo was confermed by RT/PCR and immunohistochemical staining using anti-HCV core monoclonal antibody. These HCV-expressing model rats will be useful for determining the pathological role of HCV in vivo.

由于缺乏丙型肝炎病毒(HCV)感染的小动物模型，阻碍了对该病毒发病机制的阐明。本研究的目的是建立体内基因转移表达丙型肝炎病毒的动物模型，并利用其研究肝损伤和肝癌的发生机制。在本研究中，我们使用了阳离子脂质体介导和去唾液酸糖蛋白受体介导的体内基因转移。以β-肌动蛋白启动子驱动LacZ基因的pActLacZ为报告载体，用X-Gal组织化学方法证实了该方法的可行性。对于丙型肝炎病毒的表达，使用了由β-肌动蛋白启动子驱动的丙型肝炎病毒基因组全长编码序列的表达载体。用抗丙型肝炎病毒核心单抗进行免疫组织化学染色和RT/PCR法检测大鼠肝细胞内丙型肝炎病毒的表达。这些表达丙型肝炎病毒的模型大鼠将有助于确定丙型肝炎病毒在体内的病理作用。

项目成果

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