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

[目的]本研究的目的是通过移植增殖性肝细胞并应用胶原酶基因反义核酸治疗肝硬变。[材料和方法]研究过程如下。1)建立小鼠和大鼠的肝硬变模型，分析肝纤维化的发生机制。B)可增殖的肝细胞杂交瘤是用栓性肝细胞、实质肝细胞和非实质肝细胞组成的杂交瘤。C)将杂交瘤移植到肝硬变小鼠的肝脏内，并将反义的胶原酶mRNA注入肝脏。对再生正常肝脏的条件进行了研究。[结果]用CCl4复制大鼠肝纤维化模型，观察羟丙醇在大鼠肝硬变过程中的变化。并研制了一种新的激光选择性细胞融合装置。利用该装置，我们尝试制作了实质肝细胞和栓塞型肝细胞的杂交瘤，该杂交瘤可以增殖。我们利用仙台病毒和磷脂制备了hVJ脂质体，用于胶原酶-mRNA反义基因的转导，在分析肝纤维化过程中的研究仍在进行中。

[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.