核因子-κB在白细胞介导大鼠移植胰腺缺血再灌注损伤中的作用

An essential role for NF-kappa B in neutrophil-dependent ischemia/reperfusion injury following pancreaticoduodenal transplantation in rats

目的探讨核因子（NF）-κB在大鼠移植胰腺再灌注损伤中的作用及其可能机制。方法应用同系大鼠异位全胰十二指肠移植模型，大鼠移植术前和术后5min静脉注射NF-κB抑制剂脯氨酸二硫代氨基甲酸酯（ProDTC）（15mg／kg），移植术后24h经腹主动脉取血测定大鼠血清肿瘤坏死因子（TNF）-α、巨噬细胞炎性蛋白（MIP）-2浓度酶联免疫吸附试验（ELISA）、血糖、淀粉酶、脂肪酶水平。测定胰腺组织NF-κBp65蛋白含量（Westernblot）、TNF-αmRNA、MIP-2mRNA、细胞间黏附分子（ICAM）-1mRNA表达逆转录-聚合酶链反应（RT—PCR）、髓过氧化物酶（MPO）活性。并进行组织学观察。结果移植后24hProDTC组和对照组血清中TNF-α、MIP-2水平、胰腺组织中NF-κBp65蛋白含量、TNF-α mRNA、MIP-2mRNA、ICAM-1mRNA表达、MPO活性均升高，而ProDTC组水平明显低于对照组（P〈0．05）。移植后24h血糖、脂肪酶水平在ProDTC组[（9．1±2．8）mmol／L，（192±26）U／L]明显低于对照组[（13．0±3．1）mmol／L，（297±31）U／L，P〈0．05]。ProDTC组胰小叶间质水肿和胰小叶内中性粒细胞浸润较轻。结论移植胰腺缺血再灌注后，NF—κB活化上调了TNF-α、MIP-2、ICAM-1表达从而增加中性粒细胞浸润，外源性应用NF-κB抑制剂ProDTC可以减轻移植胰腺缺血再灌注损伤。

Objective To investigate the effects of nuclear factor-κB （NF-κB） on reperfusion injury following pancreaticoduodenal transplantation in rats. Methods The （homologous） male Wistar rat model of heterotopic total pancreaticoduodenal transplantation was used. The proline dithiocarbamates （ProDTC）-treated rats received the intravenous injection of ProDTC （15 mg/kg） 5 min before and after operation. The content of NF-κBp65 in pancreas nuclear extracts （Western blot） ,the expression of TNF-α, MIP-2 and ICAM-1 transcripts （RT-PCR）, the concentrations of serum TNF-α and MIP-2 （ELISA）, blood sugar （BG） and serum concentrations of amylase and lipase, and pancreas myeloperoxidase （MPO） were analyzed. Results The content of NF-κBp65, the transcription of TNF-α, MIP-2 and ICAM-1, and the release of serum TNF-α and MIP-2 were significantly increased after transplantation both in the untreated and ProDTC-treated rats, while the levels in the ProDTC-treated group were obviously higher than in the untreated group. Administration of ProDTC resulted in a statistically significant decrease in blood sugar [ （9.1 ＋2.8） mmol/L] and serum concentration of lipase [ （ 192 ＋26） U/L] against the levels in the untreated rats [ （13.0 ＋3.1） mmol/L, （297 ＋31） U/L,P 〈0.05]. Coneltmion NF-KB inhibitor ProDTC can decrease the amount of NF-κB, and inhibit the elevation of TNF-α, MIP-2 and ICAM-1 and early neutrophil accumulation in the pancreas. NF-κB inhibitor ProDTC has protective effects on the pan- creatic ischemia/reperfusion injury after pancreaticoduodenal transplantation.