核因子-κB抑制剂对大鼠胰十二指肠移植后缺血-再灌注损伤的保护作用

Protective effects of nuclear factor-κB inhibitors on ischemia/ reperfusion injury in rat pancreaticoduodenal transplantation

目的:探讨大鼠胰十二指肠移植后核因子(NF)-κB活化对炎性介质表达和中性粒细胞浸润集聚的影响。方法:建立大鼠腔静脉内分泌引流、肠道外分泌引流的动物模型,脯氨酸二硫代氨基甲酸酯(ProDTC)组供体胰腺保存在ProDTC的UW液中,受体鼠于移植前30 min腹腔注射ProDTC(15 mg/kg);对照组供体胰腺仅保存在UW液中,受体鼠腹腔注射等量生理盐水,分别于再灌注后第0、1、3、6、12、24 h等时点,处死大鼠,取血后切取移植胰腺。采用Western-blot法检测不同组胰腺组织中NF-κBp65蛋白的表达,逆转录-聚合酶链反应(RT-PCR)法检测肿瘤坏死因子-α(TNF-α)、巨噬细胞炎性蛋白-2(MIP-2)、胞间黏附分子-1(ICAM-1)mRNA表达,检测胰腺组织髓过氧化物酶(MPO)的活性,酶联免疫吸附试验(ELISA)检测血清TNF-α和MIP-2的含量。结果:与对照组相比,ProDTC组NF-κB p65蛋白和TNF-α、MIP-2、ICAM-1 mRNA表达明显下降,MPO活性明显减低(P均<0.01)。结论:ProDTC抑制NF-κB活化,下调TNF-α、MIP-2、ICAM-1的表达,从而减轻中性粒细胞浸润及胰十二指肠移植后的缺血再灌注损伤。

Objective： To discuss the effect of nuclear factor-κB（NF-κB） activating on cytokines expression and neutrophil accumulation in rat pancreaticoduodenal transplantation.Methods： To establish a rat model of pancreaticoduodenal transplantation with venacava endocrine drainage and enteric eccrine drainage.The recipient rats were introperitoneally injected with 15mg/kg of proline dithiocarbamate（ProDTC）in experiment group and with equal amount of normal saline in control group 30 minutes before transplantation.The donor pancreases were kept in UW solution containing ProDTC in experiment group and UW solution alone in control group for 12 hours.At 0,1,3,6,12,24 h after transplantation,animals were sacrificed at each time point in each group.The pancreatic grafts were removed for MPO,and NF-κB p65 protein and TNF-α,MIP-2,ICAM-1 mRNA analysis by using Western blot and RT-PCR assay,and the TNF-α,MIP-2 analysis in serus by using ELISA methods.Results：Compared with NS control rats the level of NF-κ B p65 significantly decreased suppressing the transcription of TNF-α,MIP-2 and ICAM-1 and the release of serum TNF-α and MIP-2 as well as the activity of myeloperoxidase（MPO） in rats receiving ProDTC（all P 〈 0.01）.Conclusion： ProDTC protects against pancreasticoduodenal ischemia-reperfusion injury by suppressing NF-κ B activation and subsequent expression of proinflammatory mediators.