Melby K, Toews G B, Pierce A K
Am Rev Respir Dis. 1985 Apr;131(4):559-63. doi: 10.1164/arrd.1985.131.4.559.
Elastase activity generated during lung defense against aerobic bacteria was studied in an animal model. Bronchoalveolar lavage (BAL) fluid from hamsters inoculated with bacteria was assayed for elastase activity at 0, 2, 4, 6, and 8 h after inoculation using a synthetic substrate of elastase, succinyl-trialanine-nitroanilide (SLAPN). Streptococcus pneumoniae type 25 inoculation led to a peak elastase activity of 0.72 +/- 0.27 X 10(-3) units, not significantly different from baseline (0.41 +/- 0.08 X 10(-3) units) or saline control (0.33 +/- 0.18 X 10(-3) units). In contrast, inoculation with Pseudomonas aeruginosa strain PAO-1 (a species known to produce elastase as well as other virulence factors) produced peak elastase activity of 3.0 +/- 1.2 X 10(-3) units in BAL fluid, significantly higher than either pneumococcus type 25 or saline control (p less than 0.025). Inoculation with Pseudomonas aeruginosa strain E-64, an isogenic mutant of PAO-1 that produces a nonfunctional elastase, led to peak levels similar to the PAO-1 strain, suggesting that the presence of bacterial elastase was not the primary factor in BAL fluid elastase activity. Total numbers of granulocytes in BAL fluid from pneumococcus-inoculated animals (144 +/- 31 X 10(6] was significantly higher (p less than 0.05) than from either the PAO-1 (74 +/- 31 X 10(6] or E-64 (99 +/- 27 X 10(6] strains of Pseudomonas, Use of selective enzyme inhibitors of elastase, diisopropyl fluorophosphate and disodium ethylenediaminetetraacetate, implied that the majority of elastase activity in BAL fluid was due to a serine protease, of which granulocyte elastase is the primary source.(ABSTRACT TRUNCATED AT 250 WORDS)
在一个动物模型中研究了肺部抵御需氧菌过程中产生的弹性蛋白酶活性。对接种细菌的仓鼠的支气管肺泡灌洗(BAL)液,在接种后0、2、4、6和8小时使用弹性蛋白酶的合成底物琥珀酰 - 丙氨酰 - 硝基苯胺(SLAPN)测定弹性蛋白酶活性。接种25型肺炎链球菌导致弹性蛋白酶活性峰值为0.72±0.27×10⁻³单位,与基线(0.41±0.08×10⁻³单位)或生理盐水对照(0.33±0.18×10⁻³单位)无显著差异。相比之下,接种铜绿假单胞菌PAO - 1菌株(一种已知会产生弹性蛋白酶以及其他毒力因子的菌种)在BAL液中产生的弹性蛋白酶活性峰值为3.0±1.2×10⁻³单位,显著高于25型肺炎球菌或生理盐水对照(p<0.025)。接种PAO - 1的同基因突变体、产生无功能弹性蛋白酶的铜绿假单胞菌E - 64菌株,导致峰值水平与PAO - 1菌株相似,这表明细菌弹性蛋白酶的存在不是BAL液中弹性蛋白酶活性的主要因素。接种肺炎球菌的动物的BAL液中粒细胞总数(144±31×10⁶)显著高于铜绿假单胞菌的PAO - 1(74±31×10⁶)或E - 64(99±27×10⁶)菌株(p<0.05)。使用弹性蛋白酶的选择性酶抑制剂二异丙基氟磷酸酯和乙二胺四乙酸二钠表明,BAL液中大部分弹性蛋白酶活性归因于一种丝氨酸蛋白酶,其中粒细胞弹性蛋白酶是主要来源。(摘要截短于250字)