支气管哮喘和慢性阻塞性肺疾病患者诱导痰中基质金属蛋白酶及其抑制剂与气道炎症及气流受限

Metalloproteinase-9/tissue inhibitor of metalloproteinase-1 in induced sputum in patients with asthma and chronic obstructive pulmonary disease and their relationship to airway inflammation and airflow limitation

目的探讨支气管哮喘(简称哮喘)和慢性阻塞性肺疾病(COPD)患者诱导痰中基质金属蛋白酶9(MMP-9)和基质金属蛋白酶抑制剂1(TIMP-1)的水平及其与炎性细胞数、肺功能的关系。方法分别选择14例缓解期哮喘患者(哮喘组)、12例稳定期 COPD 患者(COPD 组)和10名健康对照者(健康对照组)进行肺功能测定和用诱导痰检查方法对痰炎性细胞进行分类计数,并用酶联免疫吸附试验(ELISA)法测定诱导痰上清液中白细胞介素4(IL-4)、MMP-9和 TIMP-1浓度。结果哮喘组患者诱导痰中嗜酸粒细胞、中性粒细胞分别为0.181±0.067、0.30±0.07,健康对照组为0.007±0.005、0.26±0.06,COPD 组为0.042±0.017、0.50±0.10,3组细胞间比较差异有统计学意义(F 值分别为4.32、4.13,P 均<0.05)。哮喘组、COPD 组、健康对照组间诱导痰中 IL-4浓度分别为(19±7)×10^(-3)g/L、(14±6)×10^(-3)g/L、(11±4)×10^(-3)g/L,3组诱导痰中 IL-4浓度比较差异无统计学意义(F=1.56,P 均>0.05),且分别与嗜酸粒细胞、中性粒细胞和第一秒用力呼气容积占预计值百分比(FEV_1占预计值%)无相关(r 分别为0.33、0.11、0.19、0.25、0.39、0.40、0.21、0.35、0.17,P 均>0.05)。哮喘组和 COPD 组诱导痰中 MMP-9、TIMP-1浓度分别为(15.9±6.0)g/L、(13.4±5.1)g/L、(19.8±8.5)g/L、(16.7±7.6)g/L,健康对照组分别为(1.8±1.1)g/L、(1.3±0.9)g/L,两组MMP-9、TIMP-1浓度比较差异有统计学意义(F 值分别为2.99、4.22,P 均<0.05)。哮喘组 MMP-9浓度与嗜酸粒细胞呈正相关(r=0.71,P<0.05);COPD 组 MMP-9浓度与中性粒细胞呈正相关(r=0.59,P<0.05),但与 FEV_1占预计值%和第一秒用力呼气容积/用力肺活量(FEV_1/FVC)无相关(r 分别为0.22、0.16、0.25、0.30,P 均>0.05)。哮喘组和 COPD 组 TIMP-1浓度均与嗜酸粒细胞和中性粒细胞无相关(r 分别为0.27、0.31、0.20、0.35,P 均>0.05),但与 FEV_1占预计值%呈负相关(r 分别为-0.58、-0.62,P 均<0.05)。哮喘组和 COPD 组诱导痰中 MMP-9/TIMP-1比值分别为0.8±0.7、0.8±0.6,两组比较差异无统计学意义(F=1.78,P>0.05),但与健康对照组(1.5±0.6)比较差异有统计学意义(F=3.70,P<0.05),且与 FEV_1占预计值%呈正相关(r 分别为0.56、0.61,P 均<0.05)。结论哮喘组和 COPD 组患者诱导痰中 MMP-9/TIMP-1比值的失衡与气道炎症和气流受限有关,这种失衡在哮喘和 COPD 细胞外基质的重塑和气流受限的发病机制中发挥重要作用。

Objective To investigate the levels of metaUoproteinase-9 （MMP-9） and tissue inhibitor of metalloproteinase-1 （ TIMP-1 ） in induced sputum in patients with asthma and chronic obstructive pulmonary disease （COPD）, and its relationship to the number of inflammation cells and lung function. Methods Fourteen patients with asthma in remission stages, 12 patients with stable COPD and 10 normal control subjects were included in this study. Lung function was measured. Induced sputum was obtained and processed for cell differential and the supernatant was assayed for the concentrations of interleukin-4 （ IL-4 ）,MMP-9 and TIMP-1 by enzyme-linked immunosorbent assay （ELISA）. Results The percentage of eosinophils in induced sputum in asthmatics（0. 181 ± 0. 067） was significantly higher than that in normal control subjects （0. 007 ±0. 005） and in COPD （0. 042 ±0. 017, F=4. 32,P 〈0. 05）. The percentage of neutrophils in induced sputum in patients with COPD （0. 500 ±0. 101） was significantly higher than that in asthmatics （0. 30 ± 0. 07 ） and in normal control subjects （ 0. 26 ± 0. 06, F = 4. 13, P 〈 0. 05 ）. The concentrations of IL-4 in asthmatics, COPD and normal control subjects [ respectively, （ 19 ± 7 ） × 10 ^-3 g/L, （ 14 ± 6 ） × 10^ -3 g/L, （ 11 ± 4） × 10^ -3 g/L ] did not show significant difference （ F = 1.56, all P 〉 0. 05 ） and did not correlate with the number of eosinophils （r =0. 33,0. 11,0. 19, all P 〉0.05） and neutrophil （ r = 0. 25,0. 39,0. 40, all P 〉 0. 05 ） and FEV1 values （ predicted r = 0. 21, 0. 35, 0. 17, all P 〉 0. 05 ）. The concentrations of MMP-9 and TIMP-1 in induced sputum in asthmatics [ （ 15.9 ± 6. 0） g/L, （ 19. 8 ± 8.5） g/L, respectively] and COPD [（13.4 ±5.1） g/L, （16.7±7.6） g/L, respectively] were significantly higher than those in normal control subjects [ （ 1.8 ±1.1 ） g/L, （ 1.3 ±0. 9） g/L, respectively] （ F = 2. 99,4.22 ,respectively, all P 〈 0. 05 ）. Increased concentration of MMP-9 correlated positively with the percentage of eosinophils in asthmatics （r =0. 71, P 〈0. 05） and with the percentage of neutrophils in COPD（ r = 0. 59, P 〈 0. 05 ）, but did not correlate with FEV1 values （ predicted r = 0. 22,0. 16, all P 〉 0. 05） and FEV1/FVC （r =0. 25,0. 30, all P 〉0. 05）. Increased concentration of TIMP-1 did not correlate with the number of eosinophils （ r = 0. 27,0. 31, all P 〉 0.05 ） and neutrophil （ r = 0. 20,0. 35, all P 〉 0.05） in asthmatics and COPD, but correlated inversely with FEV1 values （predicted, respectively, r = - 0. 58, - 0. 62, all P 〈 0. 05 ）. The ratio of MMP-9/TIMP-1 was significantly lower in asthmatics 0. 8 ± 0. 7 and COPD 0. 8 ± 0. 6 than that in normal control subjects （ 1.5 ± 0. 6, F = 3.70, P 〈 0. 05 ）. The ratio was not statistically different between asthmatics and COPD （ F = 1.78, P 〉 0. 05 ）. In asthmatics and COPD patients, the ratio of MMP-9/TIMP-1 in induced sputum correlated positively with FEV1 % （respectively, r = 0. 56,0. 61, all P 〈 0. 05 ）. Conclusion An imbalance between MMP-9 and TIMP-1 in induced sputum in asthmatics and COPD is associated with airway inflammation and airflow limitation, which may play a role in the pathogenesis of extracellular matrix remodeling and airflow limitation.