基质金属蛋白酶在大鼠慢性阻塞性肺疾病模型气道细胞外基质重塑中的作用  

The role of matrix metalloproteinases in extracellular matrix remodelling in chronic obstructive pulmonary disease rat models

作　　者：李红梅[1] 崔德健[1] 佟欣[1] 马楠[1] 高亚兵[2] 崔雪梅[2] 陆连荣[1] 王德文[2] 梁延杰[1] 

机构地区：[1]解放军第304医院呼吸科,北京100037 [2]解放军第304医院军事医学科学院病理科,北京100037

出　　处：《中华内科杂志》2002年第6期393-398,共6页Chinese Journal of Internal Medicine

摘　　要：目的　研究基质金属蛋白酶 (MMPs)及其组织抑制剂 (TIMP 1)在大鼠慢性阻塞性肺疾病 (COPD)模型气道和肺组织中的表达及其在细胞外基质重塑中的作用。方法　采用熏香烟加气管注内毒素法 ,建立大鼠COPD模型 ,观察其气道重塑的病理改变、肺功能及血气变化 ;用生化法测定支气管肺组织羟脯氨酸含量 ;用免疫组化法观察MMP 9、MMP 2及TIMP 1的蛋白定位及表达 ;用逆转录 聚合酶链反应法测定MMP 9、MMP 2及TIMP 1mRNA表达 ;用SDS PAGE明胶酶谱学测定支气管肺组织MMPs酶活性。结果　用熏香烟加气管注内毒素法建立的大鼠COPD模型 ,其病理形态学改变、肺功能及血气变化均与人类COPD的改变相似。COPD模型组支气管肺组织羟脯胺酸含量、支气管黏膜下成纤维细胞、淋巴细胞数和肺泡巨噬细胞数及以Ⅰ型胶原为主的细胞外基质含量显著高于健康对照组 (P值均 <0 .0 0 1)。COPD模型组MMP 9、MMP 2及TIMP 1在气道上皮、成纤维细胞、肺泡巨噬细胞、血管内皮细胞及部分肺泡壁细胞表达均明显增强 ,支气管肺组织MMP 9、MMP 2及TIMP 1mRNA表达亦显著增强 ,72 0 0 0MMP 2及 92 0 0 0MMP 9酶活性亦显著增高。结论　MMPs表达增强提示细胞外基质降解增加 ,支气管肺结构破坏增加。TIMP 1在抑制MMPs活性的同时 ,促进成纤维细胞增生及胶原?Objective To study the role of metalloproteinases(MMPs) in the airway extracellular matrix(ECM)remodelling of chronic obstructive pulmonary disease (COPD) rat models. Methods The COPD rat models which was established by intratracheal instillation of 200 μg lipopolysaccharide once for every two weeks(twice), and exposed to 5% smoke for 0.5 h/d for 4 weeks. The pathological changes were observed, lung function and blood gas changes were also determined. The fibroblasts, lymphocytes of bronchial walls and alveolar macrophages were counted. The hydroxyproline of bronchial lung tissue homogenates were determined by biochemistry method. The expression of MMP 9,MMP 2 and tissue inhibitor of metalloproteinase 1(TIMP 1) in bronchi and lung tissue was verfied by immunohistochemical analysis and by reverse transcription polymerase chain reaction analysis. The gelatinolytic activities of MMPs of lung tissue were performed by gelatin zymographic analysis. Results The pathological changes of bronchi and lung tissue, the changes of lung function and blood gas analysis were similar to those of the COPD patients. The number of fibroblasts, lymphocytes and alveolar macrophages of model group were significantly increased than those of control group( P <0.001). The hydroxyproline of model group was significantly increased than that of control group( P <0.001). By using image analyzer, immunoreactivity of MMP 9, MMP 2,TIMP 1 were markedly increased in epithelial cells of bronchi, fibroblasts, macrophages, endothelial cells and pneumocytes in model group as compared with those of control group. The protein expressions of MMP 9,MMP 2 and TIMP 1 in model group were significantly increased than those in control group( P <0.000 1 or P <0 01). The mRNA expression of MMP 2,MMP 9 and TIMP 1 in COPD model group ( 1.11±0.06,1.04±0.26 and 0.85±0.34,respectively)were significantly increased than those in control group (0.30±0.17,0.36±0.09 and 0.23±0.08,respectively) as well( P <0.00

关 键 词：基质金属蛋白酶 慢性阻塞性肺疾病 气道细胞外基质重塑 

分 类 号：R563.9[医药卫生—呼吸系统]