NF-κB圈套技术在小鼠重症急性胰腺炎并发肺损伤中的实验研究

摘要

目的 探讨NF-κB圈套技术对小鼠重症急性胰腺炎(SAP)肺损伤的影响.方法 36只BALB/c雌性小鼠随机分为正常对照组、SAP组、NF-κB decoy处理组、错配decoy处理组,每组9只.除正常对照组外,其他组采用雨蛙素联合脂多糖改良法制备小鼠SAP肺损伤模型.制模12 h后处死,采用EMSA法、Western印迹法检测肺脏NF-κB活性和P65蛋白含量的变化;采用逆转录-聚合酶联反应(RT-PCR)法测定肺脏TNF-α、ICAM-1、IL-1 mRNA表达;同时行肺脏病理学损伤评分.结果 12 h后急性胰腺炎组和错配decoy处理组NF-κB活性较正常对照组和圈套技术处理组显著增高;然而除正常对照组外,其他三组间细胞核中NF-κB P65蛋白含量表达没有显著差异;正常对照组下游调控因子TNF-α(0.362±0.043)、ICAM-1(0.198±0.065)、IL-1 mRNA(0.321±0.089)低表达;急性胰腺炎组和错配decoy处理组下游调控因子TNF-α(1.131±0.066;1.031±0.058)、ICAM-1(0.365±0.051;0.385±0.050)、IL-1(0.869±0.110;0.961±0.061)mRNA水平升高(P<0.05);圈套技术处理组下游调控因子TNF-α(0.329±0.059)、ICAM-1(0.186±0.086)、IL-1(0.318±0.136)mRNA水平较上述两组降低(P<0.05).急性胰腺炎组和错配decoy处理组,肺脏组织水肿、炎性浸润方面的病理损伤评分高于正常对照组和圈套处理组(P<0.05).结论 应用圈套技术抑制NF-κB的活性及减少下游炎症介质的表达,可减轻SAP并发的肺组织损伤.%Objective To investigate the effect of nuclear faetor-kappB decoy oligodeoxynucle-prides on severe acute pancreatitis complicated with lung injury. Methods Thirty-six female BALB/C mice were randomized into the normal group, SAP group, NF-κB decoy treated group and NF-κB scramble treated group. Severe acute pancratitis complicated lung injury was induced by intraperitoneal injection of cerlein (50 μg/kg/h) for 4 h and administration of intraperitoneal lipopolysaccharide (LPS 6 mg/kg) 6 h after the initial cerulean injection. Twelve hours after LPS injection, we detected nuclear factor-kappB/DNA binding activation and content, examined the mRNA expression of tumor necrosis factor (TNF-α), interleukin-1 (IL-1) and adhesion molecule-1 (ICAM-1) in lung tissues and assessed the score of histopathology of lung. Results EMSA showed transfection of NF-κB decoy ODN, but not of scrambled decoy DON, via intravenous injection resulted in a marked reduction in the binding of NF-κB nearly to the control group's level. But western blot showed that the content of NF-κB of the decoy treated group was almost the same as that of the other groups. RT-PCR indicated that the ex-pression of mRNA of TNF-α, IL-1 and ICAM-1 increased significantly in SAP group as compared with the normal group. Transfection of NF-κB decoy but not of its scrambled form resulted in a significant inhibition of overexpression of TNF-α, ICAM-1 and IL-1. In the SAP and the scrambled treated group, histological damage in lungs included wall thickening, inflammatory infiltration and hemor-rhage. These changes were strongly eliminated by the introduction of NF-κB decoy but not scrambled ODN. Conclusion With suppressed activation of NF-κB and decreased expression of the inflammatory cytokine, NF-κB decoy ODN can relieve acute pancreatitis complicated with lung injury.