Zhang Xiao-jun, He Shao-heng, Xie Hua, Wei Ji-fu, Liu Chang-ting
Respiratory Department of South Building, General Hospital of PLA, Beijing 100853, China.
Zhonghua Jie He He Hu Xi Za Zhi. 2004 Oct;27(10):649-53.
To investigate the correlations between chymase activity, levels of tryptase, interleukin-8 (IL-8), eotaxin, neutrophil and eosinophil numbers in the sputum of elderly patients with chronic obstructive pulmonary disease (COPD) and their clinical implications.
A total of 73 elderly patients with COPD (21 severe, 21 moderate and 31 mild) were recruited into the study. IL-8 and eotaxin levels were detected with Sandwich enzyme-linked immunosorbent assay (ELISA) procedures and tryptase levels were measured with antibody against tryptase with Uni CAP system. Chymase activity was determined spectrophotometrically (410 nm) by the rate of hydrolysis of 0.7 mmol/L N-succinyl-L-Ala-L-Ala-L-Pro-L-Phe-p-nitroanilide (SAAPP) in NaCl 1.5 mol/L and Tris 0.3 mol/L The specific activity of chymase was expressed as mU/ml. Actions of protease inhibitors on chymase activities were also examined with enzyme assay.
(1) At acute attack stage, the sputum levels of tryptase of patients with severe, moderate and mild COPD were 284.0, 215.0 and 59.5 ng/L, respectively. The sputum levels of tryptase in the severe and moderate patients were greater than that in mild patients (P <0.01). The tryptase levels in the sputum of all patients were significantly decreased following treatment( 151.0, 92.0, 3.3 ng/L respectively; P <0. 01). At acute attack stage, the sputum levels of IL-8 and eotaxin were 1 299. 8,454. 9,78.7 ng/L; 22.7, 15.1, 7.4 ng/L, respectively. Severe and moderate-patients were greater than that of patients at mild stages. At convalescent stage,the sputum levels of IL-8 and eotaxin were 1 037. 5,326.6, 67.9 ng/L; 7.9, 6.3, 6.8 ng/L, respectively. The IL-8 and eotaxin levels in the sputum of severe and moderate patients were significantly decreased following treatment (P < 0.01). (2) The specific chymase activities of severe and moderate patients were higher than mild patients. Native protease inhibitors alpha1-antitrypsin (alpha1-AT) and soy bean trypsin inhibitor (SBTI) inhibited 83% and 89% enzymatic chymase activity, respectively. (3) The numbers of neutrophils and eosinophils in sputum were increased in patients with COPD at the acute stage compared with the mild convalescent stage. At all remission stages, the elevated numbers of cells were significantly reduced. (4) At acute attack stage of patients with COPD, there were correlations between the sputum levels of tryptase, IL-8, eotaxin, neutrophils and eosinophils; and there were correlations between the sputum levels of tryptase, chymase and neutrophils.
COPD is not merely a neutrophil associated disease, but mast cells, eosinophils and their released mediators also actively contribute to the pathogenesis of the disease.
探讨老年慢性阻塞性肺疾病（COPD）患者痰液中糜酶活性、类胰蛋白酶、白细胞介素-8（IL-8）、嗜酸性粒细胞趋化因子水平、中性粒细胞及嗜酸性粒细胞数量之间的相关性及其临床意义。
共纳入73例老年COPD患者（重度21例、中度21例、轻度31例）。采用双抗体夹心酶联免疫吸附测定（ELISA）法检测IL-8和嗜酸性粒细胞趋化因子水平，用Uni CAP系统的抗类胰蛋白酶抗体检测类胰蛋白酶水平。通过测定在1.5 mol/L氯化钠和0.3 mol/L Tris中0.7 mmol/L N-琥珀酰-L-丙氨酸-L-丙氨酸-L-脯氨酸-L-苯丙氨酸-对硝基苯胺（SAAPP）的水解速率，用分光光度法（410 nm）测定糜酶活性。糜酶的比活性以mU/ml表示。还用酶法检测了蛋白酶抑制剂对糜酶活性的作用。
（1）在急性发作期，重度、中度和轻度COPD患者痰液中类胰蛋白酶水平分别为284.0、215.0和59.5 ng/L。重度和中度患者痰液中类胰蛋白酶水平高于轻度患者（P<0.01）。所有患者治疗后痰液中类胰蛋白酶水平均显著降低（分别为151.0、92.0、3.3 ng/L；P<0.01）。在急性发作期，痰液中IL-8和嗜酸性粒细胞趋化因子水平分别为1 299.8、454.9、78.7 ng/L；22.7、15.1、7.4 ng/L。重度和中度患者高于轻度患者。在恢复期，痰液中IL-8和嗜酸性粒细胞趋化因子水平分别为1 037.5、326.6、67.9 ng/L；7.9、6.3、6.8 ng/L。重度和中度患者治疗后痰液中IL-8和嗜酸性粒细胞趋化因子水平显著降低（P<0.01）。（2）重度和中度患者的糜酶比活性高于轻度患者。天然蛋白酶抑制剂α1-抗胰蛋白酶（α1-AT）和大豆胰蛋白酶抑制剂（SBTI）分别抑制糜酶活性83%和89%。（3）与轻度恢复期相比，COPD患者急性期痰液中中性粒细胞和嗜酸性粒细胞数量增加。在所有缓解期，升高的细胞数量均显著减少。（4）在COPD患者急性发作期，痰液中类胰蛋白酶、IL-8、嗜酸性粒细胞趋化因子、中性粒细胞和嗜酸性粒细胞水平之间存在相关性；痰液中类胰蛋白酶、糜酶和中性粒细胞水平之间存在相关性。
COPD不仅是一种与中性粒细胞相关的疾病，肥大细胞、嗜酸性粒细胞及其释放的介质也在该疾病的发病机制中发挥积极作用。
