供肝自然杀伤细胞通过上调精氨酸酶-1表达减轻肝移植急性排斥反应  

Donor liver natural killer cells alleviate acute liver graft rejection by up-regulation expression of arginase-1

作　　者：黄永亨 钟东佳[1] 龙天柱[2] 俞建东[3] 林泽宇 李文滨[4] 万云乐[3] 

机构地区：[1]揭阳市人民医院肝胆外科 [2]广州市儿童医院乳腺外科 [3]中山大学附属第六医院肝胆外科 [4]中山大学附属孙逸仙纪念医院肝胆外科

出　　处：《中华实验外科杂志》2014年第2期301-303,共3页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金资助项目（30671987）

摘　　要：目的 探讨供体肝脏自然杀伤（NK）细胞减轻大鼠肝移植急性排斥反应的作用机制.方法 以Lewis大鼠为肝移植供体、BN大鼠为受体,采用改良“二袖套”法建立大鼠肝移植模型,并应用60Co射线照射清除供体免疫细胞以及经门静脉回输供肝NK细胞等技术,设立3个实验组：A：急性排斥组（Lewis-BN）,n=18;B：单纯60Co照射排斥组（Lewis-BN＋供体60Co照射）,n=18;C：60Co照射＋供肝NK门静脉回输排斥组（Lewis-BN＋供体60Co照射＋受体门静脉输注供肝NK细胞）,n=18.分别于术后第7天收集外周血清,采用酶联免疫吸附试验（ELISA）法检测转化生长因子-β1（TGF-β1）分泌水平,同时切取移植肝行病理学检查,Western blot法检测移植肝精氨酸酶-1（Arg-1）蛋白的表达.结果 与A组比较,B组移植肝组织急性排斥反应程度明显加重,外周血TGF-β1分泌水平[（28.00±6.08） μg/L比（11.00±0.57） μg/L）,P＜0.01]和移植肝组织Arg-1表达[（0.687±0.040）比（0.487 ±0.030）,P＜0.05]均降低;但是,经供肝NK细胞门静脉输注后的C组移植肝组织急性排斥反应程度较B组明显减轻,外周血TGF-β1分泌水平[（11.00±0.57）μg/L比（27.60±1.45） μg/L,P＜0.05]和移植肝组织Arg-1表达[（0.487±0.030）比（0.605±0.080）,P＜0.05]均升高.结论 供肝NK细胞可以减轻大鼠肝移植术后急性排斥反应,其机制可能与Arg-1蛋白的表达上调有关.Objective To investigate the machenism of donor liver natural killer（NK） cells alleviating actue rejection after liver transplantation.Methods Lewis rats wereused as liver graft donors and BN rats as recipients,＂Lewis-BN＂ as acute rejection model was established by the modified two-cuff method.The donor leukocytes were depleted by total body irradiation of 60Co source,and the donor passenger leukocyte immunity were reconstituted by transfusion donor liver NK cells via portal vein during the course of transplantation of the irradiate liver.Three various transplantation groups were designed：Group A.acute rejection group （Lewis-BN）,n =18,Group B.60Co irradiation acute rejection group （60Co-Lewis-BN）,n =18,C.60Co irradiation ＋ donor liver NK cells transfusionvia portal vein acute rejection group （60CoLewis-BN ＋ donor NK cells）.The recipients were sacrificed day7 posttransplantation respectively,TGF-β1level in peripheral blood,arginase-1 （Arg-1） protein expression in liver tissues and histo pathological of liver grafts were observed and detected.Results Acute rejection in B group was more severe than A group,peripheral blood TGF-β1 levels [（28.00 ± 6.08） μg/L vs.（11.00 ± 0.57） μg/L,P 〈 0.01] and Arg-1 expression （0.687 ± 0.040 vs.0.487 ± 0.030,P 〈 0.05） of the liver in group B were significantly lower than group A; But compared with B group,the C group alleviate the actue rejection donor following liver NK cells transfution via portal vein,and the peripheral blood TGF-β1 levels [（11.00 ±0.57） μg/L vs.（27.60 ± 1.45） μ.g/L,P 〈 0.05] and Arg-1 expression （0.487 ± 0.030 vs.0.605 ± 0.080,P 〈 0.05）of the liver in group C group were significantly lower than group B.Conclusion Donor liver NK cells can alleviate liver allograft acute rejection,the machenism may be associated with up-regulation expression of Arg-1 in the liver.

关 键 词：供肝自然杀伤细胞 精氨酸酶-1 急性排斥反应 肝脏移植 大鼠 

分 类 号：R657.3[医药卫生—外科学]