Department of Molecular Microbiology and Immunology, Johns Hopkins Bloomberg School of Public Health, Baltimore, Maryland 21231, USA.
J Appl Physiol (1985). 2013 Jun;114(11):1637-44. doi: 10.1152/japplphysiol.00026.2013. Epub 2013 Apr 4.
Intratracheal elastase challenge of laboratory animals has long been established as a model for observing the physiological and morphological changes that result from alveolar destruction, the hallmark of emphysema. However, instillation of elastase suspended in buffer results in widespread inflammation and variable emphysematous lesions, which has made the identification of specific cellular and molecular events associated with the onset of emphysema difficult to define. Here we establish a bead-based elastase delivery system that induces localized tissue destruction, a key event in the initiation of emphysema. Elastase was coupled to bisacrylamide beads, which were shown to retain enzymatic activity prior to intratracheal administration in mice. C57BL/6 mice were given a single dose of 40,000 beads, which became distributed throughout the small airways and parenchyma of the lung. Elastase-coupled beads resulted in a quantifiable loss of alveolar tissue immediately surrounding the beads, an effect that was not observed with beads that lacked protein altogether or with beads containing elastase inactivated by an irreversible inhibitor. Furthermore, beads bound with active elastase elicited local recruitment of mononuclear cells, including macrophages, and polymorphonuclear neutrophils to the site of bead deposition, a feature consistent with the cellular infiltration observed following conventional solubilized elastase challenges. This work identifies a novel bead-based enzyme delivery system that also extends the elastase model of emphysema to permit the characterization of mechanisms that drive alveolar surface area loss following elastin degradation in focal emphysematous lesions.
气管内弹性蛋白酶挑战已被广泛应用于实验室动物模型,用于观察肺泡破坏(肺气肿的标志)导致的生理和形态学变化。然而,弹性蛋白酶在缓冲液中的注入会导致广泛的炎症和可变的肺气肿病变,这使得确定与肺气肿发作相关的特定细胞和分子事件变得困难。在这里,我们建立了一种基于珠粒的弹性蛋白酶传递系统,该系统可诱导局部组织破坏,这是肺气肿发生的关键事件。弹性蛋白酶与双丙烯酰胺珠粒偶联,在给予小鼠气管内给药之前,这些珠粒显示出保持酶活性。给予 C57BL/6 小鼠 40,000 个珠粒的单剂量,这些珠粒分布在小气道和肺实质中。弹性蛋白酶偶联珠粒导致紧邻珠粒的肺泡组织发生可量化的损失,而完全缺乏蛋白质的珠粒或含有通过不可逆抑制剂失活的弹性蛋白酶的珠粒则没有观察到这种效果。此外,与缺乏活性的弹性蛋白酶结合的珠粒会引起单核细胞(包括巨噬细胞和多形核中性粒细胞)局部募集到珠粒沉积部位,这一特征与传统可溶性弹性蛋白酶挑战后观察到的细胞浸润一致。这项工作确定了一种新型基于珠粒的酶传递系统,该系统还扩展了肺气肿的弹性蛋白酶模型,从而可以对弹性蛋白降解导致局灶性肺气肿病变中肺泡表面积损失的机制进行特征分析。