Mechanisms of neutrophil damage to human alveolar extracellular matrix: the role of serine and metalloproteases.
Many syndromes of lung injury are associated with accumulation of neutrophils within the pulmonary parenchyma. These neutrophils have the capacity to produce lung injury by products including proteases and reactive oxygen species (ROS). We examined the ability of activated neutrophils to solubilize human alveolar extracellular matrix (ECM), and by use of scavengers and inhibitors, evaluated the role of ROS and proteases in this process. Supernatants of phorbol myristate acetate-activated neutrophils routinely solubilized 10.2% +/- 0.8% (n = 30) of collagen in human alveolar ECM, as measured by hydroxyproline release. Scavengers of ROS had no significant effect on ECM solubilization. Inhibitors of metalloproteases partially inhibited ECM solubilization (38.5% +/- 4.6% inhibition by ethylenediaminetetraacetic acid [n = 6], and 37.0% +/- 14.7% by 1,10-phenanthroline [n = 6]; p less than 0.05). Inhibitors of the neutrophil serine proteases, elastase and cathepsin G, markedly inhibited ECM solubilization (100.9% +/- 3.7% by alpha 1-protease inhibitor [alpha 1-PI] [n = 6] and 81.9% +/- 0.1% by soybean trypsin inhibitor [n = 6]; p less than 0.01). Since alpha 1-PI completely inhibited solubilization, metalloprotease activity appeared to be related to serine protease activity. This finding was confirmed by the observation that addition of a metalloenzyme activator, p-aminophenylmercuric acetate, in the presence of alpha 1-PI, restored solubilization to the same level as that inhibited by metal chelators. We conclude that human neutrophil metalloproteases and serine proteases directly solubilize human alveolar ECM. Furthermore, neutrophil serine proteases activate latent metalloproteases. However, ROS were not demonstrated to play a major role in ECM solubilization in our system.
Palmgren
M S
MS
Department of Medicine, Tulane University School of Medicine, New Orleans, La.
deShazo
R D
RD
Carter
R M
RM
Zimny
M L
ML
Shah
S V
SV
D001485:Basement Membrane / Q000187:drug effects; D003094:Collagen / Q000378:metabolism; D005109:Extracellular Matrix / Q000187:drug effects* / Q000378:metabolism; D006801:Humans; D007678:Kidney Glomerulus; D008163:Luminescent Measurements; D008666:Metalloendopeptidases / Q000502:physiology; D009504:Neutrophils / Q000502:physiology*; D017830:Octoxynol; D011092:Polyethylene Glycols / Q000494:pharmacology; D011650:Pulmonary Alveoli / Q000502:physiology; D012128:Respiratory Distress Syndrome / Q000097:blood*; D012694:Serine / Q000502:physiology; D012995:Solubility
D011092:Polyethylene Glycols; D012694:Serine; D017830:Octoxynol; D003094:Collagen; D008666:Metalloendopeptidases