Abstract
目的 通过观察活体或冷保存期离体门静脉灌注供肝转染白细胞介素-1受体相关激酶-4(IRAK-4)特异短发夹RNA(shRNA)对再灌注后受体TNF-α产生的影响,判断以IRAK-4为肝移植缺血再灌注损害(I/RI)治疗靶点的可行性并探索可行的shRNA治疗途径.方法雄性SD大鼠,随机分为冷缺血转染组、活体转染组、对照组,以两袖套法建立同种异体肝移植模型.冷缺血转染组于冷缺血期经门静脉灌注转染携带染IRAK-4-shRNA的转染质粒pSIIRAK-4;活体转染组在门静脉袖套吻合完成后,经门静脉分支注入pSIIRAK-4;对照组不予任何处理.按门静脉血流恢复后第0 min、60 min及180 min分为三个亚组,逆转录-聚合酶链式反应及蛋白免疫印记法测定肝组织的染IRAK-4 mRNA和蛋白表达水平;酶连免疫吸附法检测肝组织NF-κB活性及血清TNF-α含量.结果再灌注后活体转染组、对照组的IRAK-4蛋白与mRNA表达水平、NF-κB活性以及TNF-α含量均高于冷缺血转染组(P＜0.01);冷缺血转染组的IRAK-4表达明显抑制,再灌注后各时点差异无显著性(P＞0.05).结论以IRAK-4为靶点的冷缺血shRNAs转染途径能有效减轻肝移植时的I/RI,但能否以IRAK-4作为预防其他器官的I/RI的治疗靶点尚需深入研究。
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