A DEVELOPMENT OF THE NEW METHOD WHICH ACCELERATES LIVER REGENERATION BY USING ADENOVIRUS EXPRESSION VECTOR SYSTEM

利用腺病毒表达载体系统加速肝脏再生新方法的开发

• 批准号: 09470262
• 负责人: KOGURE Kimitaka
• 金额: $ 8.64万
• 依托单位: Gunma University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 1997
• 资助国家: 日本
• 起止时间: 1997 至 1999
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09470262/
• 关键词: activin A; follistatin; TGF-β; liver regeneration; adenovirus vector; dominant negative receptor of activin A; gene transfer; virus vector; TGF-β受容体; ペプチド合成

Augmentation of liver regeneration, with an artificial support to maintain liver function necessar for survival, would be beneficial for the treatment of fulminant hepatic failure caused by viral infection and hepatic toxins. As well, massive hepatectomy to resect liver tumors can be performed safely if the growth of the remnant liver is promoted effectively. Recently, the roles of various growth factors in the regulation of hepatocyte growth have been elucidated. Activin A which belongs to the transforming growth factor β super gene family is a potent autocrine growth inhibitor in hepatocytes. Activin A is induced in vivo after partial hepatectomy. Low levels of the messenger RNA for the bA-subunit of activin are markedly increased 20 hours after hepatectomy. When follistatin, which binds activin and blocks its action, is infused into the portal vein immediately after hepatectomy of 70 %, DNA is synthesized several hours earlier than in control rats and liver regeneration is significa … More ntly augmented. Follistatin provides a potential therapeutic approach to promoting liver regeneration because it specifically blocks autocrine growth inhibitor. The results were reported in Gastroenterology (vol 108 : 1136-1142, 1995). Then, we obtained the results that the intravenous administration of follistatin effectively promoted liver regeneration after partial hepatectomy. These were reported in Hepatology (vol : 24 : 361-366, 1996). We then investigated the effects of follistatin on liver regeneration under more critical condition, 90 % hepatectomy in rats and obtained the results that follistatin was quite effective in promoting liver regeneration (J Hepatol 29: 977-984, 1998). This raises the possibility that liver regeneration may be accelerated by the internal blockade of the autocrins of the autocrine actions of activin and TGF-β. The present study was conducted to address this issue. For this purpose, we constructed the dominant negative activin and TGF-β receptors on the hepatocytes. The adenovirus (5-dlX) vector was transferred with dominant negative activin and TGF-β type II receptors and cultured in the 293 cells. The virus vector was directly injected via the portal vein into the liver of rats and DNA synthesis was analyzed by measuring labeling index of BrDU and the DNA content. As a control, adenovirus which was transferred lacZ gene was also injected directly via the portal vein into hepatocytes and the β-galactosidase activity was analyzed by measuring the staining index of 5-bromo-4-chloro-indolyl-b-D-galactoside. We are investigating the effects of the dominant negative activin and TGF-β type II receptors expressed on the hepatocytes for liver regeneration. Less

肝脏再生的增强，并提供人工支持以维持生存所必需的肝功能，将有益于治疗由病毒感染和肝毒素引起的暴发性肝衰竭。同样，如果有效促进残余肝脏的生长，可以安全地进行大规模的肝术以恢复肝肿瘤。最近，已经阐明了各种生长因子在肝增长调节中的作用。属于转化生长因子β超基因家族的激活素A是肝细胞中潜在的自分泌生长抑制剂。部分肝术后体内诱导活化素A。肝切开术后20小时，用于激活素的BA亚基的信使RNA水平较低。当肝蛋白结合并阻断其作用的卵泡素在70％的肝术后立即被感染到门静脉中时，与对照大鼠相比，DNA合成了几个小时，而肝脏再生则是显着的……更加迅速地增强。 Follistatin提供了一种潜在的治疗方法来促进肝脏再生，因为它专门阻止了自分泌抑制剂。结果报道了胃肠病学（第108：1136-1142、1995）。然后，我们获得了结果，即肝静脉曲张素的静脉内给药可在部分肝切开术后有效促进肝脏再生。这些在肝病学中报道（第24卷：361-366，1996）。然后，我们研究了卵泡素对肝脏再生的影响，在更为危急的情况下，大鼠的肝术90％，并获得了follistatin在促进肝脏再生方面非常有效的结果（J HEPATOL 29：977-984，1998）。这增加了肝脏再生的可能性，可以通过激活素和TGF-β的自分泌作用的内部阻断来加速肝脏再生。本研究是为了解决这个问题。为此，我们在肝细胞上构建了主要的阴性激活素和TGF-β受体。腺病毒（5-DLX）载体用显性阴性激活素和TGF-βII型受体转移，并在293个细胞中培养。通过门静脉直接注入病毒载体进入大鼠的肝脏，并通​​过测量BRDU和DNA含量的标记指数来分析DNA合成。作为对照，也直接通过门静脉直接注入了转移的LACZ基因的腺病毒，并通过测量5-溴-4-氯-4-氯 - 氯米 - indolylyl-b-d-galactoside的染色指数来分析β-半乳糖苷酶活性。我们正在研究在肝细胞上表达的主要阴性激活素和TGF-βII型受体的作用。较少的

项目成果

Kimitaka Kogure: "Immediate onset of DNA synthesis in remnant rat liver after 90% hepatectomy by an administration of follistatin" Journal of Hepatology. 29. 977-984 (1998)

Kimitaka%20Kogure:%20“立即%20onset%20of%20DNA%20synthesis%20in%20remnant%20rat%20liver%20after%2090%%20hepatection%20by%20an%20administration%20of%20follistatin”%20Journal%20of%20Hepatology.%

Kimitaka kogure: "Uric acid changes in serum during different forms of hepatic vascular inflow occlusion"Life Science. 60(20). 1781-1791 (1997)

Kimitaka kogure：“不同形式的肝血管流入阻塞期间血清中尿酸的变化”生命科学。

Kimitaka Kogure: "A comparative study of the anatomy of rat and human livers"Journal of Hepatobiliary Pancreatic Surgeries. 6. 171-175 (1999)

Kimitaka Kogure：“大鼠和人类肝脏解剖学的比较研究”肝胆胰外科杂志。

Kimitaka Kogure: "Evaluation of serum uric acid changes in different forms of hepatic vascular inflow occlusion in human liver surgeries"Life Science. 64(5). 305-313 (1999)

Kimitaka Kogure：“人类肝脏手术中不同形式的肝血管流入阻塞中血清尿酸变化的评估”生命科学。

Kimitaka Kogure: "Uric acid chages in serum during different forms of hepatic vascular inflow occlusion" Life Science. 60(20). 1781-1791 (1997)

Kimitaka Kogure：“不同形式的肝血管流入阻塞期间血清中的尿酸发生变化”生命科学。