CAJ | 학술논문

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.
목적 탐토공체간장자연살상(NK)세포감경대서간이식급성배척반응적작용궤제.방법 이Lewis대서위간이식공체、BN대서위수체,채용개량“이수투”법건립대서간이식모형,병응용60Co사선조사청제공체면역세포이급경문정맥회수공간NK세포등기술,설립3개실험조:A:급성배척조(Lewis-BN),n=18;B:단순60Co조사배척조(Lewis-BN+공체60Co조사),n=18;C:60Co조사+공간NK문정맥회수배척조(Lewis-BN+공체60Co조사+수체문정맥수주공간NK세포),n=18.분별우술후제7천수집외주혈청,채용매련면역흡부시험(ELISA)법검측전화생장인자-β1(TGF-β1)분비수평,동시절취이식간행병이학검사,Western blot법검측이식간정안산매-1(Arg-1)단백적표체.결과 여A조비교,B조이식간조직급성배척반응정도명현가중,외주혈TGF-β1분비수평[(28.00±6.08) μg/L비(11.00±0.57) μg/L),P＜0.01]화이식간조직Arg-1표체[(0.687±0.040)비(0.487 ±0.030),P＜0.05]균강저;단시,경공간NK세포문정맥수주후적C조이식간조직급성배척반응정도교B조명현감경,외주혈TGF-β1분비수평[(11.00±0.57)μg/L비(27.60±1.45) μg/L,P＜0.05]화이식간조직Arg-1표체[(0.487±0.030)비(0.605±0.080),P＜0.05]균승고.결론 공간NK세포가이감경대서간이식술후급성배척반응,기궤제가능여Arg-1단백적표체상조유관.
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.