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

背景。已知抗炎细胞因子白介素-1受体拮抗剂（IL-1Ra）可减轻肝缺血-再灌注损伤。因此，我们希望研究IL-1Ra基因转染大鼠肝脏对肝缺血再灌注损伤的影响。通过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入门静脉，将IL-1Ra cDNA送入大鼠肝脏。基因传递后24小时，大鼠肝脏局部缺血90分钟后再灌注。再灌注180分钟取肝组织和血清，检测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-1Ra基因递送对非缺血肝叶在再灌注后立即部分切除7天生存率的影响。在两种递送方法中，IL-1Ra基因转移导致血清IL-1Ra浓度显著升高，在递送后24小时达到最高水平。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。在给予IL-1Ra的大鼠中，在再灌注180分钟时，肝脏损伤以及促炎细胞因子的产生以循环IL-1Ra蛋白的浓度依赖性方式显着减少。经腺病毒载体基因转染的大鼠7天存活率高于对照动物。IL-1Ra基因进入肝脏可能用于治疗肝移植后肝缺血再灌注损伤。

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.