急性肺损伤大鼠肺部细胞因子水平变化及肺泡上皮细胞的凋亡情况  

Changes of pulmonary cytokine levels and apoptosis of alveolar epithelial cells in acute lung injury rats

作　　者：宋小莲[1] 白冲[2] 傅佩芳[1] 颜正茂[1] 张国良[1] 彭爱梅[1] 王昌惠[1] 

机构地区：[1]同济大学附属第十人民医院呼吸科,上海200072 [2]第二军医大学附属长海医院呼吸科

出　　处：《上海医学》2010年第11期1027-1030,I0001,共5页Shanghai Medical Journal

基　　金：国家自然科学基金项目资助(30670913)

摘　　要：目的观察急性肺损伤(ALI)时肺部几个主要凋亡相关细胞因子的水平变化及其与肺泡上皮细胞凋亡的相关性。方法采用高浓度氧吸入、脂多糖(LPS)及盐酸(HCL)气道注入等方法制作ALI模型,酶联免疫吸附法检测大鼠肺泡灌洗液(BALF)中表面活性物质A(SPA)、血管紧张素Ⅱ(AngⅡ)、凋亡蛋白-1(Fas)、凋亡蛋白-1配体(FasL)、肿瘤坏死因子(TNF)-α等的水平,免疫组织化学法检测肺组织中Fas/FasL的表达,原位缺口末端标记(TUNEL)法检测肺泡上皮细胞的凋亡情况。同时使用FasL干预体外培养的Ⅱ型肺泡上皮细胞,观察其凋亡情况。结果各组动物模型上表现出的肺组织有不同程度ALI病理表现,其中LPS2组最典型。与对照组相比,LPS1、LPS2、HCl、高浓度氧组的BALF中SPA均显著降低(P值均<0.01),Fas、FasL均显著升高(P值分别<0.05、0.01)。LPS2组肺组织标本Fas和FasL的阳性表达率分别为(24.7±4.8)%、(17.9±5.2)%,均显著高于对照组的(2.7±0.7)%、(2.3±0.8)%(P值均<0.05)。用FasL处理RLE-6TN细胞24 h后,有(39.2±6.5)%的细胞出现调亡。结论多种细胞因子在ALI肺部炎症中发挥作用,其中Fas/FasL介导的肺泡上皮细胞凋亡是ALI的重要致伤因素。Objective To observe the changes of several major apoptosis-related cytokines in the lung and its correlation with apoptosis of alveolar epithelial cells（AECs） in acute lung injury（ALI）.Methods Rat ALI models were induced by hyperoxia,infusion of lipopolysaccharide or hydrochloric acid via airway.The levels of surfactant protein A（SPA）,AngiotensinⅡ,Fas,FasL and tumor necrosis factor-α（TNF-α） in the alveolar lavage fluid（BALF） were assayed by ELISA.The apoptotic AECs were detected by in situ nick end-labeling（TUNED technique,and the pulmonary expression of Fas/FasL was detected by immunohistochemisty staining. Furthermore,the apoptosis ofⅡtype alveolar epithelial cells pretreated with FasL was also detected.Results Classic pathological changes of ALI of different degrees were found in all animal groups,especially in LPS2 group. Compared to control group,SPA concentrations in BALF were significantly lower（P0.01） and Fas/FasL was significantly higher（P0.05,0.01） in the LPS1,LPS2,HCI,and high oxygen groups.The positive rates of Fas and FasL in LPS 2 group（[24.7±4.8]%and[17.9±5.2]%respectively） were significantly higher than those of the control group（[2.7±0.7]%and[2.3±0.8]%,respectively,P0.05）.The apoptotic rate of RLE-6TN cells was（39.2±6.5）%24 h after treatment with FasL.Conclusion Multiple cytokines may be involved in the lung inflammation of ALI.Apoptosis of AECs mediated by Fas/FasL may play an important role in the development and progression of ALI.

关 键 词：急性肺损伤 肺泡上皮细胞 凋亡 凋亡蛋白-1/凋亡蛋白-1配体 

分 类 号：R563[医药卫生—呼吸系统]