Gene therapy-for live cirrhosis by multiplicable hepatocyte and antisense of collagenase-mRNA

基因治疗-通过增殖性肝细胞和胶原酶-mRNA反义治疗活肝硬化

• 批准号: 07671353
• 负责人: OHKOHCHI Nobuhiro
• 金额: $ 0.38万
• 依托单位: Tohoku University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1995
• 资助国家: 日本
• 起止时间: 1995 至 1996
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07671353/
• 关键词: Liver cirrhosis; Hepatocyte transplantation; Laser device; Gene therapy; Antisense therapy; レーザー; 遺伝子治療

[Purpose] The purpose of this study is to establish a therapy for the liver cirrhosis by a transplantation of proliferative hepatocyte and administration of antisense for mRNA of collagenase. [Materials and Methods] Process of the study were as followed. A) Model of liver cirrhosis is made in mouse and rat and mechanism of fibrosis is analyzed. B) Hybridomas of hepatic cells which can proliferate are made with embrional hepatocytes and parenchymal and nonparenchymal hepatocytes. C) The hybridomas are transplanted into the cirrhotic liver and antisense for the collagenese mRNA is also injected the liver. Conditions of regenerating normal liver are studied. [Results] We made liver cirrhosis using CCl_4 and examined hidroxyprolil activity during changing liver cirrhosis in rats. And we developped a new device for selective cell fusion by Laser device. Using this device we tried to make hybridoma of parenchymal hepatocyte and embrional hepatocyte, which can proliferate. We prepared HVJ liposome using Sendai virus and phospholipid for transfection of antisense of collagenase-mRNA.Studies of investigation in analysis of fibrosing process of liver were still remained.

[目的]建立肝细胞移植联合胶原酶反义基因治疗肝硬化的新方法。[材料与方法]研究过程如下。A）在小鼠和大鼠中制备肝硬化模型并分析纤维化的机制。B）用胚胎肝细胞以及实质和非实质肝细胞制备可增殖的肝细胞杂交瘤。C）将杂交瘤移植到患肝中，并将胶原mRNA的反义基因也注射到肝中。研究了正常肝再生的条件。[结果]采用四氯化碳（CCl_4）诱导大鼠肝硬化模型，并在肝硬化过程中检测羟脯氨酸的活性。并研制了一种新型的激光选择性细胞融合装置。利用该装置，我们尝试了制备具有增殖能力的肝实质细胞和胚性肝细胞杂交瘤。我们用仙台病毒和磷脂制备了HVJ脂质体，用于转染胶原酶-mRNA的反义核酸，并对肝纤维化过程进行了分析。

项目成果

板垣英雄、大河内信弘、他: "レーザーを用いた新しい選択的細胞融合法の開発" 薬理と臨床. 6. 1535-1538 (1996)

Hideo Itagaki、Nobuhiro Okochi 等人：“使用激光开发新的选择性细胞融合方法”药理学和临床研究 6. 1535-1538 (1996)。

Saya K,Ohkohchi N,Mori S: "Changes in the ability of ATD synthesis in the mitochondrial membrane in the rat liver injured carbon tetrachloride." Tohoku Exp. J.Med.Vol178. 425-430 (1996)

Saya K、Ohkohchi N、Mori S：“四氯化碳损伤大鼠肝脏线粒体膜中 ATD 合成能力的变化。”

Itagaki H.,et al: "Selective selection method by Laser de vice" Acta Histochemica et Cytochemica. vol29sup. 750-751 (1996)

Itagaki H.等人：“激光装置的选择性选择方法”Acta Histochemica et Cytochemica。

板垣英雄、他: "レーザーを用いた新しい選択的細胞融合法の開発" 薬理と臨床. vol6,8. 1535-1538 (1996)

Hideo Itagaki 等人：“使用激光开发新的选择性细胞融合方法”，药理学和临床研究，第 6 卷，1535-1538（1996 年）。

Seyak,Ohkohchi N,et al: "Changes in the ability of ATP synthesis in the mltochondrial membrane in the rat liver in jured by carbon tetrachloride" Tohoku J Exp Med. 178. 425-430 (1996)

Seyak，Ohkohchi N，等人：“四氯化碳损伤后大鼠肝脏线粒体膜中 ATP 合成能力的变化”Tohoku J Exp Med。