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目的探讨核因子κB(NF-κB)活化对急性坏死性胰腺炎(ANP)肺泡巨噬细胞(AM)分泌炎症介质的影响和对肺损伤的影响。方法30只成年SD大鼠随机分为正常对照组、ANP后1、3、6、12 h组,每组6只。逆行性胰胆管注射5％牛磺酸钠建立ANP大鼠模型,正常对照组大鼠自胆胰管内逆行注入生理盐水。经支气管肺泡灌洗获取肺泡巨噬细胞,检测支气管肺泡灌洗液(BALF)中蛋白含量、肺组织髓过氧化物酶(MPO)水平、肺泡巨噬细胞分泌肿瘤坏死因子α(TNFα),一氧化氮(NO)水平,测定肺泡巨噬细胞TNFαmRNA、诱导型一氧化氮合酶(iNOS)mRNA表达情况。行肺组织的病理学检查并评分,同时行免疫组化检查肺组织中NF-κB的表达。结果正常肺组织较少见到NFκB活化的AM,ANP各组肺组织均可见到NF-κB活化,并且随时间进展NF-κB逐渐由细胞质更多进入细胞核。ANP大鼠肺损伤随着病情进展而逐渐加重,肺组织MPO及支气管肺泡灌洗液中蛋白含量逐渐升高;肺泡巨噬细胞分泌TNFα,NO水平逐渐升高,至6 h达到高峰,12 h又回落。ANP发生后,肺泡巨噬细胞TNFαmRNA、iNOSmRNA的表达情况与TNFα,NO的变化趋势相似。ANP大鼠各组指标与正常对照组相比均有统计学差异(P<0．05)。结论ANP形成后,NF-κB发生活化,使AM炎症介质表达增强,并促进肺损伤。
Objective To investigate the effect of nuclear factor κB (NF-κB) activation on inflammatory mediators secreted by alveolar macrophages (AM) in acute necrotizing pancreatitis (ANP) and its effect on lung injury. Methods Thirty adult SD rats were randomly divided into normal control group, 1, 3, 6, 12 h groups after ANP. The ANP rat model was established by injecting 5% sodium taurinate into the pancreaticobiliary duct. Rats in the normal control group were injected with normal saline retrogradely into the pancreatic duct. Alveolar macrophages were obtained by bronchoalveolar lavage. Protein levels in bronchoalveolar lavage fluid (BALF), levels of myeloperoxidase (MPO) and alveolar macrophages were assayed for the secretion of tumor necrosis factor α (TNFα) Nitric oxide (NO) was measured to determine the mRNA expression of TNFαmRNA and iNOS in alveolar macrophages. The pathological examination of lung tissue was performed and scored. Meanwhile, the expression of NF-κB in lung tissue was examined by immunohistochemistry. Results NF-κB activation was not found in normal lung tissue. Activation of NF-κB was observed in lungs of ANP groups. NF-κB gradually increased from cytoplasm to nucleus. The lung injury in ANP rats gradually aggravated with the progression of the disease. The content of protein in lung tissue MPO and bronchoalveolar lavage fluid gradually increased. The levels of TNFα and NO secreted by alveolar macrophages increased gradually and peaked at 6 h, Down again. After ANP, the expression of TNFαmRNA and iNOSmRNA in alveolar macrophages were similar to that of TNFα and NO. ANP rats in each group indicators compared with the normal control group were statistically significant (P <0.05). Conclusion After the formation of ANP, NF-κB is activated and the expression of AM inflammatory mediators is enhanced and lung injury is promoted.