The effect of simultaneous transfection of HGF gene-plasmid and NFκB-decoy using ultrasound exposure with contrast agent in a rat kidney allograft model

我们之前已经证明，在慢性同种异体移植肾病（CAN）的发展机制中，肾移植过程中由于缺血-再灌注损伤导致的功能肾小球数量减少以及急性移植排斥反应是高度相关的；核因子κ b （NFκB）的活化、粘附分子表达的增强和细胞因子的产生，是该过程中的重要因素。与Morishita， Tomita等人一起，我们先前开发了“NFκB诱饵”，即针对NFκB的顺式元件寡聚脱氧核糖核酸诱饵，它调节炎症细胞因子和粘附分子的表达，并通过超声心动图造影剂暴露将其转染到急性排斥反应的大鼠肾移植模型中（Gene Ther.2003 Mar; 10(5):415-25）。在本研究中，基于我们对诱导肾小管移植失败的原因的调查结果，我们考虑了超声造影剂、Optison和超声暴露可能直接损害肾小管的可能性，并进行了实验来减少这种不良影响。为了通过同时抑制超声暴露引起的急性移植物排斥反应和组织损伤，以及减轻肾移植过程中的缺血再灌注损伤，诱导移植物永久存活，我们开发了一种保护肾小管损伤的肝细胞生长因子（HGF）基因质粒，并将其与nf κ b诱饵转染到供肾中。(我们之前描述了HGF对肾小管损伤的抑制作用。参见Azuma等人：J.Am Soc Nephrol；田中等人：美国J移植。)目前在急性排斥反应的大鼠肾移植模型中获得的数据表明，与单独转染诱饵的对照组相比，同时转染HGF基因质粒和nfκ b诱饵的动物移植存活时间明显延长。不幸的是，治疗未能诱导永久性移植物存活，但我们将进一步研究开发永久性移植物存活的肾移植方法。少

We have previously demonstrated that in the mechanism of development of chronic allograft nephropathy (CAN), a decreased number of functioning glomeruli due to ischemic-reperfusion injury during renal transplantation as well as acute graft rejection are highly involved ; and activation of nuclear factor kappaB (NFκB) and enhancement of expression of adhesion molecules and cytokine production, mediated by such a factor, constitute important factors in that process. Together with Morishita, Tomita and others, we previously developed "NFκB-decoy", decoy cis-elements oligo deoxyribonucleic acid against NFκB, which modulates expression of inflammatory cytokines and adhesion molecules, and transfected them into kidney transplants in a rat kidney transplantation model for acute rejection using ultrasound exposure with an echocardiographic contrast agent (Gene Ther.2003 Mar ; 10(5):415-25). In the present study, based on the results from our investigation for causes of the failure to induce pe … More rmanent graft survival, we considered the possibility that the renal tubules may directly be impaired by echocardiographic contrast agent, Optison, and ultrasound exposure, and conducted experiments to reduce such adverse effects. In order to induce permanent graft survival by simultaneously suppressing acute graft rejection and tissue damage caused by ultrasound exposure with Optison, as well as reducing ischemic-reperfusion injury during renal transplantation, we developed a plasmid of the gene for hepatocyte growth factor (HGF), which provides protection against renal tubule damage, and transfected it and NFκB-decoy into the donor kidney. (We previously described the inhibitory effect of HGF on renal tubule damage. See Azuma, et al. : J.Am Soc Nephrol ; Tanaka et al. : Am J Transplant.) The present data obtained in the rat kidney transplantation model for acute rejection have shown a significantly prolonged graft survival for animals receiving simultaneous transfection of HGF gene-plasmid and NFκB-decoy, compared with the control transfected with the decoy alone. Unfortunately the treatment failed to induce permanent graft survival, but we will further investigate development of a kidney transplantation method for permanent graft survival. Less