Development of a super liver graft resistant to various kinds of injuries by a gene-transfer technique

通过基因转移技术开发出抗各种损伤的超级肝移植物

基本信息

批准号：
13357011
负责人：
SHIMADA Mitsuo
金额：
$ 32.86万
依托单位：
Kyushu University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (A)
财政年份：
2001
资助国家：
日本
起止时间：
2001 至 2002
项目状态：
已结题

项目摘要

This study was conducted to clarify the mechanism of ischemia-reperfusion (I/R) injury or small-for-size graft (SSG) injury, and to create a super liver graft resistant to various kinds of injuries using a gene-transfer technique.(1) Development of gene-transfer technique :Instead of retrovirus vector with critical risk of leukemia (Science 2002), electroporation-mediated gene transfer to cold preserved graft was carried out, using a new instrument designed for a rat liver graft. Using a combined method of electroporation and hydrodynamic (=positive pressure to the portal vein) technique, luciferase-gene expression in the graft after liver transplantation was found to be increased. But a significant graft damage was also observed. A new gene-transfer method using nano-carrier is, therefore, now applied.(2) Liver transplant (LT) model using a small-for-size graft :In a new rat LT model using small-for-size graft (SSG : 30% of whole liver volume), survival of transplanted rats was suggested to prolong either by either creation of partial porto-systemic shunt (splenopexy beneath the skin) or administration of somatostatin analogue having effect of reduced portal pressure. Therefore, excessive portal perfusion of SSG was proved to be a critical cause of SSG injury. A new LT model was, furthermore, completed in mice.(3) Role of Rho-kinage on ischemia-reperfusion injury :Rho-kinase, a molecular substance related to vascular contraction or neutrophil migration, was found to played an important role on I/R injury. Its inhibitor (Fasudil) inhibited cytokine production (TNFa and IL1b) and free radical production in I/R injury. By administration of Fasudil, postoperative elevation of AST and ALT, and necrotic areas in the specimen, were reduced, furthermore, rat's survival rate after LT improved in a 24-hour cold-preserved LT model. Either anti-sense or dominant negative of Rho-kinase gene is now applied to liver graft using a non-viral gene-transfer technique.

进行了这项研究是为了阐明缺血性灌注（I/R）损伤或小大小的移植（SSG）损伤的机制，并使用基因转移技术的基因转移技术（而不是基因转移技术的开发（而不是逆转录病毒）的转移（而不是逆转录病毒），而不是逆转录病毒，而不是逆转录病毒，则对各种类型的损伤产生了抗性的损伤。使用专为大鼠肝移植的新仪器进行冷保留的移植物。使用一种组合的电穿孔和流体动力学方法（=门静脉静脉的正压）技术，发现肝移植后移植物中的荧光素酶基因表达增加。但是也观察到了显着的移植损伤。因此，现在已经应用了使用纳米载体的一种新的基因转移方法。（2）肝移植（LT）模型，使用小型移植物使用：在新的大鼠LT模型中，使用小型移植物（SSG：SSG：SSG：30％的全肝脏体积的30％），建议通过移植的大鼠的生存来延长替代的替代，以延长两种形式的良好型，而替代了一部分（替代）。或给予生长抑素类似物，其门户压力降低。因此，事实证明，过度的SSG门户灌注是SSG损伤的关键原因。此外，在小鼠中完成了一种新的LT模型。（3）Rho-timage在缺血 - 灌注损伤上的作用：Rho-激酶是一种与血管收缩或中性粒细胞迁移有关的分子物质，在I/R损伤上起着重要作用。它的抑制剂（Fasudil）抑制了I/R损伤中的细胞因子产生（TNFA和IL1B）和自由基产生。通过施用Fasudil，AST和ALT的术后升高以及样品中的坏死区域降低，此外，在24小时冷藏的LT模型中，LT改善了LT后的大鼠存活率。现在使用非病毒基因转移技术将反义或主要的rho-激酶基因的抗渗透或显性阴性应用于肝移植。