Interleukin-1 Receptor Antagonist Gene Transfer into Rat Liver after Ischemia-Reperfusion

背景已知抗炎细胞因子白细胞介素-1受体拮抗剂（IL-1 Ra）可减少肝脏缺血-再灌注损伤。因此，我们希望研究IL-1 Ra基因导入大鼠肝脏对肝脏缺血-再灌注损伤的影响。通过使用质粒阳离子脂质体或重组腺病毒载体将转基因载体单次注射到门静脉中，将IL-1 Ra cDNA递送到大鼠肝脏中。在基因递送后24小时，使大鼠经受部分肝缺血90分钟，随后再灌注。于再灌注180 min取肝组织和血清，观察肝损伤程度及血清和组织中促炎细胞因子的表达水平。此外，我们评估了IL-1 Ra基因对再灌注后立即切除非缺血性肝叶的7天存活率的影响。在两种递送方法的情况下，IL-1 Ra的基因转移导致血清IL-1 Ra浓度显著升高，其在递送后24小时达到最大水平。然而，腺病毒载体的最高血清浓度是脂质体处理动物的1000倍。在IL-1 Ra递送的大鼠中，在再灌注180分钟时，肝损伤以及促炎细胞因子的产生以循环IL-1 Ra蛋白的浓度依赖性方式显著减少。与对照动物相比，接受腺病毒载体基因递送的大鼠具有更高的7天存活率。IL-1 Ra基因在肝内的表达可能对减轻肝移植后缺血再灌注损伤有一定的治疗作用。

Background. The anti-inflammatory cytokine interleukin-1 receptor antagonist (IL-1Ra) is known to reduce hepatic ischemia-reperfusion injury. Therefore, we wished to examine the effect of IL-1Ra gene delivery into the rat liver on hepatic ischemia-reperfusion injury.Methods. IL-1Ra cDNA was delivered into the rat liver by a single injection of the transgene vector into the portal vein using either the plasmid-cationic liposome or the recombinant adenoviral vector. At 24 hours after the gene delivery, rats were subjected to partial liver ischemia for 90 minutes followed by reperfusion. Liver tissue and serum samples were taken at 180 minutes of reperfusion, and the degree of the liver injury as well as the expression level of pro-inflammatory cytokines in the serum and tissue were investigated. In addition, we assessed the effect of IL-1Ra gene delivery on the 7-day survival rate when the nonischemic liver lobe was partially excised immediately following reperfusion.Results. In both cases of delivery methods, gene transfer of IL-1Ra resulted in significant elevation of serum IL-1Ra concentration, which reached maximal levels at 24 hours following the delivery. However, the highest serum concentration with the adenoviral vector was 1, 000-fold of that in the liposome-treated animals. In the IL-1Ra delivered rats, liver damage, as well as production of pro-inflammatory cytokines, at 180 minutes of reperfusion was significantly reduced in a concentration-dependent manner of the circulating IL-1Ra protein. Rats subjected to the adenoviral vector gene delivery had higher 7-day survival rates compared with control animals.Conclusions. IL-1Ra gene delivery into the liver may be of therapeutic use for abrogating hepatic ischemia-reperfusion injury after transplantation.