供肝转染IL-10对大鼠肝脏移植术后排斥反应的影响

Allograft transduction of IL-10 effects on rejection reaction following rat liver transplantation

目的观察供肝转染白细胞介素-10(IL-10)后基因表达,并明确IL-10对大鼠肝脏移植急性排斥反应的作用。方法在建立稳定的大鼠肝脏移植模型的基础上,应用改良“二袖套法”行Lewis到BN大鼠肝移植34例,A组12例,为对照组;B组11例,为空载体转染组,术中供肝冷保存期门静脉注射真核细胞转染剂Lipofectamine20000-pcR3.1空载体质粒复合物,保存45min后行肝移植;C组11例,为重组白细胞介素-10(RIL-10)转染组,术中供肝冷保存期门静脉注射真核细胞转染剂Lipofectamine20000-pcR3.1RIL-10复合物,保存45min后行肝移植。术后第6天,每组处死BN大鼠3只。取血清检测肝功能指标、IL-10和白细胞介素-2(IL-2)水平及RT-PCR检测肝细胞IL-10表达水平,了解肝移植排斥反应情况。结果A、B、C组大鼠术后第6天检查A、B组丙氨酸转氨酶(ALT)、天冬氨酸转氨酶(AST)、总胆红素(TBIL)、碱性磷酸酶(ALP)和IL-2水平均较C组升高,差异显著;而血白蛋白(ALB)水平则明显低于C组;检查移植肝脏为中、重度急性排斥反应。C组血清IL-10水平明显升高。肝上下腔静脉(SHVC)IL-10水平可达639.72ng/ml,是肝下下腔静脉(IHVC)IL-10水平的近1.4倍。排斥反应轻微或不明显。肝脏组织RT-PCRRIL-10表达在治疗组明显处于高水平。结论脂质体介导,经门静脉途径进行供肝转染IL-10,可以使IL-10在肝脏获得较高水平的表达。IL-10对肝移植术后的急性排斥有抑制作用,可以有效地延长移植物存活时间。脂质体介导RIL-10转染对肝移植术后的慢性排斥没有抑制作用。

Objective To investigate the effect of genetic modulation of the hepatic graft with IL-10 during liver preservation in rat liver transplantation. Methods Thirty four cases of orthotopic liver transplantation from Lewis to BN rats were performed using the two cuff technique.All rats were devided to 3 groups.Groups A(n=12)was control.Grafts were transvescularly transfected through introportal injection of Lipofectamine 2000-pcR3.1 complex (group B,n=11)or Lipofectamine 2000-pcR3.1 RIL-10 complex(group C,n=11)during cold preservayion ex tivo.Three rats of each group were killed on day 6 postoperation.Transgene expression of RIL-10 was assessed by means of RT-PCR.Histologic rejection score,serum IL-2,ALT,AST,ALP,TBIL and ALB levels were also assessed. Results Serious acute rejection was found in the liver graft of group A and B.Serum IL-2,ALT,AST,ALP and TBIL levels were higher in group A and B than group C.Levels of IL-10 from SHVC were significant higher than those from IHVC.Transgene expressions of RIL-10 in group C were higher than those of qroup A and B. Conclusion Liposome mediated gene transfection through portal vein during cold preservation ex vivo to doner liver can successfully achieve local gene exression.IL-10 gene transfer into rat liver allografts improved graft function,reduced acute rejection,downregulated serum IL-2 level,and prolonged survival of liver allograft.