摘要
目的观察氯化钆(GdCl3)诱导急性坏死性胰腺炎(ANP)肺泡巨噬细胞(AM)凋亡时核因子-κB(NF-κB)表达情况,探讨NF-κB在其中的作用及机制。方法36只SD大鼠随机分为正常对照组,ANP组,GdCl3治疗组。逆行性胰胆管注射5%牛磺胆酸钠建立ANP大鼠模型,正常对照组大鼠以同法注入生理盐水,GdCl3治疗组制模后即刻自阴茎背静脉注射GdCl3。各组大鼠于成模后6 h经支气管肺泡灌洗获取AM,检测支气管肺泡灌洗液(BALF)中TNF-α和IL-1β含量及肺组织髓过氧化物酶(MPO)水平。用琼脂糖凝胶电泳、流式细胞仪检测AM的凋亡情况;Western-blot检测AM中NF-κB活性。同时对肺组织行病理学检查。结果ANP组TNF-α和IL-1β含量高于对照组(P<0.05),而治疗组显著低于ANP组(P<0.05)。仅治疗组DNA电泳见典型的细胞凋亡的梯状条带。对照组,ANP组,治疗组AM凋亡率分别为(10.81±0.86)%,(6.47±1.52)%,(17.41±3.36)%,3组间差异均有统计学意义(P<0.05);Western-blot检测3组AM中NF-κB表达的相对灰度值分别为(0.80±0.05),(1.96±0.15),(1.42±0.10),3组间差异亦有统计学意义(P<0.05)。AM的凋亡率与NF-κB活性呈负相关(r=-0.554,P<0.01)。结论GdCl3可能通过抑制NF-κB的活化诱导大鼠ANP肺泡巨噬细胞凋亡,从而减轻肺损伤。
Objective To discuss the role of activation of nuclear factor Kappa B （ NF-κB） in apoptosis of alveolar macrophages （AM） induced by gadolinium chloride （GdCl3 ） in acute neerotizing pancreatitis （ANP）. Methods Thirty sixty adult SD rats were randomized into three groups： normal control group, ANP group and the group treated by GdCl3. ANP was induced by intraductal administration of 5 % sodium taurocholate, while the normal control rats received an infusion of normal saline. In GdCl3 treatment group, GdCl3 was injected into dorsal vein of penis fight after ANP model was established. AM were harvested by bronchoalveolar lavage six hours after model was established. The generation of TNF-α and IL-1β in bronchoalveolar lavage fluid （ BALF ） , and the level of myeloperoxidase in lung tissue were evaluated. The expression of NF-κB protein in AM was determined by western blot. The apoptosis of AM was detected by agarose gel electrophoresis and flow cytometer. The histological examination of lung tissue was checked. Results The levels of TNF-α and IL-1β in ANP group were significantly higher than the control group and GdCl3 treatment group （ P 〈 0.05 ）. Agarose gel electrophoresis showed the typical apoptotic DNA ladder only seeing in GdCl3 treatment group. The rates of apoptosis of AM in the control group, ANP group and GdCl3 treatment group were （10. 81±0. 86）% ,（6.47 ±1. 52）%, and（ 17.41±3. 36）% respectively, with statistical significance between the different groups（ P 〈 0.05 ）. The relative gray scale of NF-KB was （ 0.80±0.05 ） , （ 1.96±0. 15 ） , and （ 1.42±0. 10 ） respectively, with statistical significance between the different groups （ P 〈 0.05 ）. The apoptotic rate of AM had negative correlation （ r = - 0. 554, P = 0. 005 ）with the positive expression of NF-κB. Conclusions GdCl3 could induce the apoptosis of AM by inhibiting expression of NF-κB, and then ameliorate the lung injury of ANP rats.
出处
《中国普通外科杂志》
CAS
CSCD
北大核心
2009年第9期922-925,共4页
China Journal of General Surgery
基金
北京市科委科技新星计划资助课题(020821500190)
关键词
胰腺炎
急性坏死性
核因子-κB
氯化钆
巨噬细胞
肺泡
细胞凋亡
Panereatitis, Acute Necrotizing
Nuclear Faetor Kappa B
Gadolinium Chloride
Macrophages, Alveolar
Apoptosis
