Fera T, Abboud R T, Richter A, Johal S S
Am Rev Respir Dis. 1986 Apr;133(4):568-73. doi: 10.1164/arrd.1986.133.4.568.
In order to determine the acute effect of smoking on elastase in bronchoalveolar lavage fluid (BAL), we obtained BAL from 30 smokers twice, the first before smoking (after 8 h of abstinence) and the second 2 min to 1 h after the subjects had smoked either 2 or 4 cigarettes. Bronchoalveolar lavage was concentrated 100-fold and was assayed for elastaselike activity against succinyl-trialanyl-p-nitroanilide (SLAPN) as substrate. Elastolytic activity against insoluble 3H-elastin and immunologic neutrophil elastase levels tested with a sensitive enzyme-linked immunosorbent assay were determined in some subjects. No activity against insoluble 3H-elastin was detected using a sensitive assay capable of detecting subnanogram quantities of elastase. Total elastaselike activity in BAL against SLAPN was significantly (p less than 0.02) increased in smokers prior to smoking (47.4 +/- SD 20.2 ng human neutrophil elastase (HNE) equivalents) when compared with BAL from 7 nonsmokers (26.3 +/- SD 13.4 ng HNE); however, there was no significant change in enzyme activity in smokers' BAL after smoking. Assays with EDTA and phenyl-methyl-sulfonyl-fluoride as inhibitors suggested that approximately two thirds of elastaselike activity was due to a metalloprotease and that there was negligible serine protease activity.(ABSTRACT TRUNCATED AT 250 WORDS)
为了确定吸烟对支气管肺泡灌洗(BAL)液中弹性蛋白酶的急性影响,我们对30名吸烟者进行了两次BAL采集,第一次在吸烟前(禁欲8小时后),第二次在受试者吸2支或4支香烟后2分钟至1小时。将支气管肺泡灌洗液浓缩100倍,并以琥珀酰-丙氨酰-对硝基苯胺(SLAPN)为底物检测弹性蛋白酶样活性。在一些受试者中测定了针对不溶性3H-弹性蛋白的弹性水解活性以及用灵敏的酶联免疫吸附测定法检测的免疫中性粒细胞弹性蛋白酶水平。使用能够检测亚纳克量弹性蛋白酶的灵敏测定法未检测到针对不溶性3H-弹性蛋白的活性。与7名不吸烟者的BAL(26.3±标准差13.4纳克人中性粒细胞弹性蛋白酶(HNE)当量)相比,吸烟者吸烟前BAL中针对SLAPN的总弹性蛋白酶样活性显著增加(p<0.02)(47.4±标准差20.2纳克HNE当量);然而,吸烟者吸烟后BAL中的酶活性没有显著变化。用乙二胺四乙酸(EDTA)和苯甲基磺酰氟作为抑制剂的测定表明,大约三分之二的弹性蛋白酶样活性归因于金属蛋白酶,而丝氨酸蛋白酶活性可忽略不计。(摘要截短于250字)