基质金属蛋白酶及其组织抑制剂与慢性阻塞性肺疾病气道炎症的关系

The Relationship of Matrix metalloproteinase-9 and Its Tissue Inhibitor of Metalloproteinease-1 and Airway Inflammation of Chronic Obstructive Pulmonary Disease

目的:通过检测慢性阻塞性肺疾病(COPD)急性期痰上清液MMP-9、TIMP-1的浓度和痰液炎症细胞数目,分析MMP- 9、TIMP-1的变化与气道炎症的关系。方法:随机收集26例住院COPD急性加重期及治疗后稳定期患者和16例健康人对照组的痰,对痰细胞悬液计细胞总数,离心痰作细胞涂片瑞氏染色后行分类细胞计数;采用双抗体夹心酶联免疫吸附法检测痰上清液MMP-9、TIMP-1水平。结果:(1)COPD急性期痰液细胞总数、Neu数及Neu%分别高于稳定期和对照组,均有显著差异(均P<0.01);稳定期和对照组比较亦有显著差异(P<0.01);急性期及稳定期的Lym、Macro、Eos数均高于对照组,均有显著差异(均P<0.01);而稳定期和对照组比较无显著差异(P>0.05)。(2)急性期痰上清液MMP-9的浓度344.14 ng/ml (IQR 216.01～771.35)高于稳定期145.90ng/ml(IQR 79.85～256.40)和对照组64.37 ng/m1(IQR 30.09～82.19),均有显著差异(均P<0.01),稳定期高于对照组(P<0.01);急性期痰上清液TIMP-1的浓度94.53 ng/ml(IQR 77.60～141.09)高于稳定期61.50ng/ml(IQR 49.93～84.73)和时照组45.01 ng/ml(IQR 34.11～51.08)(均P<0.01);稳定期高于对照组(P<0.01);急性期MMP-9/TIMP-1比值3.47(IQR 2.23～7.01)高于稳定期2.32(IQR 1.32～3.13)和对照组1.24(IQR 0.77～1.53)(均P<0.01);稳定期高于对照组(P<0.01)。(3)急性期痰上清液MMP-9浓度和MMP-9/TIMP-1比值分别与Neu、Lym、Macro、Eos数比较均呈正相关(均P<0.01);而痰TIMP-1浓度分别与Neu、Lym、Macro、Eos数比较均无相关性(均P>0.05);稳定期痰上清液MMP-9浓度和MMP-9/TIMP-1比值分别与Neu、Lym、Macro、Eos数比较均无相关性(均P>0.05);TIMP-1浓度与Macro数呈正相关(r=0.487,P=0.012),而与Neu、Lym、Eos数比较均无相关性(均P>0.05)。结论:在COPD的发病中存在MMP-9/TIMP-1的失衡。MMP-9参与了COPD的气道炎症过程,在一定程度上可成为COPD急性发作的一个标志物。

Objective： To inverstigate the changes of sputum supernatant concentration of matrix metalloproteinase-9 （MMP- 9） ,tissue inhibitor of metalloproteinease-1 （TIMP-1） in the acute exacerbation of patients with chronic obstructive pulmonary disease（COPD） ,and to compare the relationship of MMP-9, TIMP-1 and inflammatory cells in the sputum. Methods： Sputum supernatants of 26 in-patients with COPD during acute exacerbation and 16 health controls were analyzed. Sputum induction was performed by inhalation of hypertonic saline solution after pre-medication with inhaled salbutamol 400μg. Sputum cells were counted after cytospin preparation and staining with Right＇s staining to assess the quality of the samples. Sputum supernatant concentration of MMP-9, TIMP-1 were measured through enzyme-linked immunosorbent assay（ELISA）. Results： （1） The total counts of sputum, neutrophils and neutrophil/leukocyte ratio （Neu/Leu%） during exacerbation in COPD were higher than that of the stable （P〈0.01） and the control （P〈0.01）. The differences between the stable and the control were significant （P〈0.01）. The counts of the lymphocyte （Lym） , the macrophage（Macro）, the eosinophil （Eos） in sputum during the acute exacerbation and the stable were higher than that of the control （P〈0.01）. （2） The sputum supernatant concentrations of MMP-9 and TIMP-1 during exacerbation of COPD were higher than that of the stable （P〈0.01） and control （P〈0.01）, and the levels of MMP-9 and TIMP-1 in stable were higher than that in control（P〈0.01）; The ratio of MMP-9/TIMP-1 during exacerbation of COPD were higher than that in stable group of COPD （P〈0.01） and control （P〈0.01）, and the ratio of MMP-9/ TIMP-1 in stable were higher than that of control（P〈0.01） ; （3） The sputum supernatant concentrations of MMP-9 and the The ratio of MMP-9/TIMP-1 during exacerbation of COPD showed positive correlations with the counts of Neu, Lyre, Macro, Eos （P〈0.01, respectively）, and there are not correlations among the concentrations of TIMP-1 and the counts of Neu, Lyre, Macro, Eos （P〉0.05, respectively）; The sputum supernatant concentrations of MMP-9 and the The ratio of MMP-9/TIMP- 1 in the stable of COPD showed no correlations with the counts of Neu、Lym、Macro、Eos （P〉0. 05 ,respectively）, there were positive correlations between the concentrations of TIMP-1 and the counts of Macro （P〈0.05） and no correlations among the concentrations of TIMP-1 and the counts of Neu, Lym, Eos （P〉0. 05 , respectively）. Conclusion：The imbalance of MMP-9/ TIMP-1 exists during the acute exacerbation of COPD , MMP-9 participates the acute airway inflammation of COPD ,and to a certain extent, sputum MMP-9 can be used as a marker of acute exacerbation in COPD.