【摘要】 目的探讨慢性阻塞性肺疾病(COPD)患者类糜蛋白酶(chymase)活性,类胰蛋白酶(tryptase)、白细胞介素8(IL8)、嗜酸粒细胞趋化因子(eotaxin)水平和中性粒细胞(NEU)、嗜酸粒细胞(EOS)计数的相关性及临床意义。方法老年COPD患者73例(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,检测诱导痰IL8、eotaxin水平。在UniCAP100全自动体外变应原检测仪上进行tryptase检测,Chymase活性测定使用琥珀酰丙氨酸丙氨酸脯氨酸苯丙氨酸酰苯氨(SAAPP)作为底物,采用酶标仪在410nm连续监测吸光度的变化。结果(1)老年急性加重期COPD患者(重、中、轻)痰tryptase的中位数分别为2840、2150、595ng/L,治疗后各组痰tryptase水平显著下降(分别为1510、920、33ng/L,P均<001)。加重期重度、中度与轻度患者比较差异均有显著性(P均=0)。急性加重期痰IL8、痰eotaxin中位数分别为12998、4549、787ng/L;227、151、74ng/L。重度、中度组痰IL8、eotaxin水平均高于轻度组(P均<001)。治疗后痰IL8、eotaxin中位数分别为10375、3266、679ng/L;79、63、68ng/L。重度、中度患者痰IL8、eotaxin水平均比治疗前显著降低(P均<001)。(2)重、中度COPD患者急性加重期痰chymase活性高于轻度组,黄豆胰蛋白酶抑制剂(SBTI)、α1抗胰蛋白酶(α1A更多还原

【Abstract】 Objective 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 Methods 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 Results (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 α 1 antitrypsin(α 1 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 Conclusion 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更多还原