脂质体介导核因子-κB诱捕物寡聚脱氧核苷酸对重症急性胰腺炎大鼠胰腺炎症因子mRNA表达和胰腺损伤的影响

Effect of Iiposome-mediated nuclear factor kB decoy oligodeoxynucleotide on the mRNA expression of inflammatory factors and injury of pancreas in rats with severe acute pancreatitis

目的:探讨脂质体(liposome)介导核因子-κB (nuclear factorκB,NF-κB)诱捕物(decoy)寡聚脱氧核苷酸(ODN)对重症急性胰腺炎大鼠胰腺NF-κB活性及受其调控炎症因子基因mRNA表达和胰腺损伤的影响．方法:除假手术组(n=10)外,其余SD大鼠以牛磺胆酸钠(STC)诱导建立重症急性胰腺炎模型后,分别于建模后1 h静脉注射裸ODN(n =10)、脂质体／decoy ODN复合物(n=10)、脂质体／scrambled ODN复合物(n=10)和生理盐水(n=10),注射4h应用电泳迁移率变动分析(EMSA)NF-κB的活性,利用逆转录－聚合酶链反应(RT-PCR)法检测胰腺组织ICAM－1,IL－1α,IL-2,TNF-α,VCAM-1 mRNA表达,同时检测血淀粉酶、胰腺组织湿／干重比率和胰腺组织髓过氧化物酶(MPO)．结果:EMSA显示,脂质体／decoy ODN复合物组NF-κB活性明显低于生理盐水组、脂质体／scrambled ODN复合物组和裸ODN组,均有显著性差异(P<0．05)．RT-PCR结果显示,脂质体／decoy ODN复合物组ICAM-1,IL-1α．IL-2,TNF-α和VCAM-1 mRNA表达低于生理盐水组、脂质体／scrambled ODN复合物组和裸ODN组,均有显著性差异(ICAM-1:0．75±0．13 vs 1．39±0．15,1.37±0．16,1.32±0．17, P<0．05;IL-1α:0．64±0．09 vs 1.34±0．20．1．30±0．14,1．25±0．20,P<0．05;IL-2:0．23±0．08 vs 0．74±0．13,0．71±0．12,0．69±0．14,P<0．05; TNF-α:0．41±0．13 vs 1．30±0．17,1．26±0．17, 1．23±0．20,P<0．05;VCAM-1:0．21±0．06vs 0．68±0．13,0．69±0．15,0．63±0．13,P<0．05)．与生理盐水组、脂质体／scrambled ODN复合物组和裸ODN组相比,脂质体／decoy ODN复合物组淀粉酶、胰腺组织湿／干重比率和胰腺组织MPO活性显著性降低(淀粉酶:5093 1．85±22432．15 nkat／L vs 188024．26±38659．56．188412．68±37988．26,183119．95±33636．23 nkat／L,all P<0．05;湿／干重比率:5．76±0．20 vs 6．77±0．18,6．72±0．18,6．35±0．12．P<0．05; MPO活性:46．68±3．00 nkat／g vs 99．02±2．50．98．1 9±2．83,98．52±2．50 nkat／g,P<0．05)．结论:NF-κB decoy ODN可特异性抑制胰腺NF-κB活性及其调控的炎症因子ICAM-1, IL-1α,IL-2,TNF-α和VCAM-1 mRNA的表达,减轻胰腺损害．

AIM： To investigate the influences of liposome-mediated nuclear factor κB （NF-κB） decoy oligodeoxynucleotide （ODN） on the NF- κB activation, inflammatory factor mRNA expression and pancreatic injury in rats with severe acute pancreatitis （SAP）. METHODS： Except for those in sham operation group, the rest rats were injected with sodium taurocholate to establish the model of SAP, and then intravenously injected with naked ODN, liposome/decoy ODN complexes, liposome/ scrambled ODN complexes, and normal saline, respectively, 1 hour later. Four hours after injection, the NF-κB activity was analyzed by electrophoretic mobility shift assay （EMSA）, and the mRNA expression of intercellular adhesion molecule-1 （ICAM-1）, interleukin-1α （IL-α）, IL-2, tumor necrosis factor-α （TNF-α） and vascular cell adhesion molecule-1 （VCAM-1） were assessed by reverse transcription-polymerase chain reaction （RT-PCR）. Meanwhile, the serum amylase level pancreatic wet/dry weight ratio and myeloperoxidase （MPO） content were detected. RESULTS： The NF-κB activity and its related inflammatory factors were observably inhibited in liposome/decoy ODN group as compared with those in normal saline group, liposom./scrambled ODN group and naked ODN group （NF-κB activation： P 〈 0.05; ICAM-1： 0.75 ± 0.13 vs 1.39 ± 0.15,1.37 ± 0.16,1.32 ± 0.17, all P 〈 0.05; IL-1α 0.64 ± 0.09 vs 1.34 ± 0.20,1.30 ± 0.14,1.25 ± 0.20, all P 〈 0.05; IL-2：0.23 ± 0.08 vs 0.74 ± 0.13, 0.71 ± 0.12, 0.69 ± 0.14, all P 〈 0.05; TNF-α： 0.41 ± 0.13 vs 1.30 ± 0.17,1.26 ± 0.17,1.23 ± 0.20, all P 〈 0.05; VCAM-1： 0.21 ± 0.06 vs 0.68 ± 0.13, 0.69 ± 0.15, 0.63 ± 0.13, all P 〈 0.05）. In comparison with normal saline, liposome/scrambled ODN and naked ODN group, the level of serum amylase, the ratio of pancreaic wet/dry weight, and the MPO content of pancreatic tissues in liposome/decoy ODN group were remarkably decreased （amylase： 50931.85 ± 22432.15 nkat/L vs 188024.26 ± 38659.56, 188412.68 ± 37988.26, 183119.95 ± 33636.23 nkat/L, P 〈 0.05; wet/dry weight ration： 5.76 ± 0.20 vs 6.77 ± 0.18, 6.72 ± 0.18, 6.35 ± 0.12, P 〈 0.05; MPO： 46.68 ± 3.00 nkat/g vs 99.02 ± Z50, 98.19 ± Z83, 98.52 ± Z50 nkat/g, V 〈 0.05）. CONCLUSION： NF-κB decoy ODN is effective in alleviating pancreatic injury during SAP through suppressing the activation of NF-κB and mRNA expression of ICAM-1, IL-1α, IL-2, TNF-1α and VCAM-1.