Induction of human airway smooth muscle apoptosis by neutrophils and neutrophil elastase.
Neutrophils are an important component of airway inflammation and may interact with human airway smooth muscle cells (HASMC). We investigated the effect of neutrophils and of neutrophil-derived proteases on HASMC survival. When co-incubated with neutrophils (0.1-1 x 10(6) cells/ml), attachment of human ASMC was reduced to 12.3 +/- 4.3% compared with untreated controls after 72 h. HASMC showed nuclear condensation and fragmentation (41.6 +/- 8.1% compared with baseline of 3.1 +/- 0.4%), and the biochemical markers of apoptosis, annexin V binding (9.7 +/- 0.7%; baseline 1.1 +/- 0.3%) and cleaved caspase-3 expression, were observed. The proteolytic activity released by neutrophils was essential for the proapoptotic effect because inhibition of elastase activity by alpha(1)-antitrypsin and MeOSuc-Ala-Ala-Pro-Ala-CMK (MSACK) reduced HASMC apoptosis. Human neutrophil elastase (0.1-3 microg/ml) induced apoptosis of HASMC, as well as other neutrophil serine proteases, cathepsin G, and proteinase 3. Fibronectin degradation products were present in HASMC supernatants exposed to neutrophil-conditioned media and to neutrophil elastase. The local release of proteases from neutrophils present in airway smooth muscle cells may lead to HASMC apoptosis as a result of matrix degradation and loss of cell attachment. This may limit pathologic changes such as ASMC hyperplasia and extracellular matrix deposition seen in airway remodeling.
Oltmanns
Ute
U
Thoracic Medicne, National Heart & Lung Institute, Imperial College, Dovehouse St., London SW3 6LY, UK.
Sukkar
Maria B
MB
Xie
Shaoping
S
John
Matthias
M
Chung
K Fan
KF
D017209:Apoptosis / Q000187:drug effects / Q000502:physiology*; D053148:Caspase 3; D020169:Caspases / Q000378:metabolism; D002478:Cells, Cultured; D018920:Coculture Techniques; D006801:Humans; D019272:Leukocyte Elastase / Q000037:antagonists & inhibitors / Q000494:pharmacology / Q000502:physiology*; D009130:Muscle, Smooth / Q000166:cytology* / Q000187:drug effects; D009504:Neutrophils / Q000166:cytology / Q000187:drug effects / Q000201:enzymology / Q000502:physiology*; D012137:Respiratory System / Q000166:cytology* / Q000187:drug effects; D015843:Serpins / Q000494:pharmacology
D015843:Serpins; D019272:Leukocyte Elastase; C505407:CASP3 protein, human; D053148:Caspase 3; D020169:Caspases