哮喘患者血浆类胰蛋白酶、IL-8和嗜酸性粒细胞趋化因子水平与类糜蛋白酶活性测定及相关性分析

Detection and correlation of tryptase, IL-8, eotaxin and chymase activity in plasma of patients with asthma

目的 :探讨哮喘患者血浆类胰蛋白酶 (tryptase)、白介素 8(IL 8)、嗜酸性粒细胞趋化因子 (eotaxin)水平和类糜蛋白酶 (chymase)活性及相关性 .方法 :分别取健康人、哮喘急性期中度和重度患者各 30例 ,静脉抽取抗凝血 .在UniCAP1 0 0全自动体外变应原检测仪上进行tryptase测定 ,采用双抗体夹心酶联免疫吸附实验 (SELISA)检测血浆IL 8,eotaxin水平 .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)做为底物 ,采用酶标仪在 4 1 0nm连续监测吸光度的变化 .结果 :急性期哮喘患者血浆tryptase(2 .8μg/L)值与对照组 (1 .2 μg/L)比较明显升高 (P <0 .0 5 ) ;IL 8值 (1 4 1 .5 μg/L)显著高于对照组 (6 .5 μg/L) (P <0 .0 5 ) ;Eotaxin值 (1 5 4 .9μg/L)显著高于对照组 (4 9.9μg/L) (P <0 .0 5 ) .1 8.3%的哮喘患者血浆chymase活性[(1 0 9.9± 1 32 .0 )nkat]高于对照组 (0nkat) ,其活性可被类胰蛋白酶抑制剂SBTI和α1 AT分别抑制 87.0 %和 88.2 % .IL 8与Eotaxin之间存在弱相关性 (r=0 .4 ,P =0 .0 4 6 ) ;Tryptase ,Eotaxin ,IL 8与血涂片中的嗜中性粒细胞、淋巴细胞、单核细胞和嗜酸性粒细胞之间均无明显相关性 .结论 :肥大细胞、中性粒细胞、嗜酸性粒细胞及其介质均参?

AIM: To investigate the correlaitons and levels of tryptase, IL 8, eotaxin and chymase activity in the plasma of patients with asthma. METHODS: A total of 60 patients with asthma at acute attack stage (30 severe and 30 moderate) were recruited into the study. IL 8 and eotaxin levels were detected by sandwich ELISA procedures and tryptase levels were measured using antibody against tryptase with UniCAP system. The chymase activity was determined spectrophotometrically (410 nm) by the rate of hydrolysis of N succinyl L Ala L Ala L Pro L Phe p nitroanilide (SAAPP) 0.7 mmol/L in NaCl 1.5 mol/L and Tris 0.3 mol/L. The actions of protease inhibitors on chymase activities were examined with enzyme assay. RESULTS: At acute attack stage, the plasma levels of tryptase (2.8 μg/L) were higher than those of healthy subjects(1.2 μg/L). The plasma levels of IL 8 (141.5 μg/L) and eotaxin (154.9 μg/L)were significantly elevated in patients with asthma than those of healthy subjects(6.5 μg/L and 49.9 μg/L, respectively, P <0.05). The specific chymase activities of 18.3 % of patients with asthma[(109.9±132.0)nkat] were higher than those of controls (0 nkat). The native protease inhibitors α1 antitrypsin (α1 AT) and soy bean trypsin inhibitor (SBTI) inhibited 87.0% and 88.2% chymase enzymatic activity, respectively. There were correlations between the levels of IL 8 and eotaxin ( r=0.4, P =0.046). There were no correlations between tryptase, IL 8, eotaxin, neutrophils, lymphocytes, monocytes and eosinophils. CONCLUSION: Mast cells, neutrophils, eosinophils and cytokines participate in the pathogenesis of allergic inflammation. Among these cells, mast cells may play a key role as they can release mediators such as tryptase, chymase, IL 8 or eotaxin which in turn recruit other inflammation cells including neutrophils and eosinophils to the involved areas.