半乳凝集素-9在诱导大鼠肝移植免疫耐受中的作用.doc

半乳凝集素-9在诱导大鼠肝移植免疫耐受中的作用 刘一鸣，何 勇，张 艳，赖星 李金政 龚建平 (400010 重庆，重庆医科大学附属第二医院肝胆外科) [摘要] 目的 通过观察半乳凝集素-9(Galectin-9)在大鼠肝移植急性排斥模型中的作用，探讨其在诱导肝移植免疫耐受中的机制。方法 建立Lewis到BN大鼠肝移植急性排斥反应模型（n=30），受体随机分为3组，每组10只。转染组于肝移植冷缺血期经门静脉转染重组galectin-9腺病毒质粒2 ml，空质粒组灌注2 ml空腺病毒载体，对照组灌注2 ml生理盐水。术后7 d各组处死5只大鼠，余观察生存率，采用Banff分级观察移植物排斥程度；实时荧光定量PCR检查肝组织Galectin-9、T-bet、RORγt mRNA表达水平；Western blot检测肝组织IFN-γ及IL-17蛋白表达水平。结果 术后7 d转染组Banff评分Ⅰ～Ⅱ级，空白质粒组及对照组为Ⅲ级；转染组、空白质粒组及对照组Galectin-9 mRNA水平依次为（1.14±0.05）、（0.46±0.03）、（0.49±0.07），转染组Galectin-9mRNA水平较其余2组明显增加(P0.05)。转染组、空白质粒组及对照组T-bet 及RORγt mRNA表达水平分别为（0.27±0.07）、（1.26±0.12）、（1.31±0.08）和（0.57±0.13）、（1.57±0.09）、（1.58±0.07），转染组T-bet及RORγt mRNA表达水平明显低于其余2组(P0.05)；转染组、空白质粒组及对照组IFN-γ及IL-17蛋白表达水平分别为（0.39±0.06）、（1.28±0.11）、（1.47±0.05）和（0.64±0.07）、（1.52±0.05）、（1.67±0.04），转染组IFN-γ及IL-17蛋白表达水平低于空白质粒组及对照组(P0.05)。结论 Galectin-9通过清除Th1/Th17细胞诱导肝移植免疫耐受形成。 [关键词] 半乳凝集素-9；肝移植；免疫耐受；大鼠 [中图法分类号] R392.4 [文献标志码] A Role of galectin-9 in inducing immune tolerance of rats after liver transplantation Liu Yiming, He Yong, Zhang Yan, Lai Xing, Li Jingzheng, Gong Jianping (Department of Hepatobiliary Surgery, Second Affiliated Hospital, Chongqing Medical University, Chongqing,400010,China) [Abstract] Objective To study the mechanism of galectin-9 in inducing immune tolerance of rats after liver transplantation by observing its role in an acute rejection model of rats. Methods An acute rejection model of Lewis-BN rats after liver transplantation was established. Thirty rats were randomly divided into transfection group, blank plasmid group, and control group (10 in each group). Rats in transfection group were injected with 2 mL recombinant Ad-galectin-9 through the portal vein, those in blank plasmid group were injected with 2 mL adenoviral vector, and those in control group were injected with 2 mL saline during the cold ischemia period. Five rats in each group were sacrificed on day 7 after operation. Survival time of the remaining rats was observed. Degree of acute graft reje