Mechanisms of extracellular matrix proteoglycan degradation by human neutrophils.
Alterations in proteoglycans (PG) located in the pulmonary interstitium may influence extracellular matrix (ECM) structure and assembly during the development of diseases in which increased numbers of neutrophils enter the lung. To evaluate potential mechanisms of PG degradation, neutrophils or purified neutrophil products were incubated with ECM that had been produced by cultured neonatal rat vascular smooth muscle cells (SMC) or lung fibroblasts (LF) and metabolically labeled with 35SO4. Matrix PG solubilization was expressed as a percentage of the spontaneous [35SO4]PG solubilization that occurred in the presence of buffer alone. Solubilization by unstimulated neutrophils was 105.8 +/- 3.1% (mean +/- SEM, n = 6) and 101.7 +/- 3.05 (n = 8) using ECM that had been produced by LF and SMC, respectively. Solubilization by neutrophils that had been stimulated with formyl-methionine-leucine-phenylalanine (FMLP) in the presence of cytochalasin B (CB) was 189.7 +/- 5.8% and 298.2 +/- 26.2% using ECM produced by LF and SMC, respectively. Matrix that had been produced by SMC was used to evaluate which neutrophil products were responsible for the degradation of PG. Addition of a specific inhibitor of neutrophil elastase (NE) to stimulated polymorphonuclear leukocytes (PMN) reduced PG solubilization by 88.3 +/- 4.8% (n = 8). Addition of an inhibitor of cathepsin G (CG), as well, did not further reduce PG degradation. Purified CG and myeloperoxidase solubilized significantly more PG, 125.8 +/- 6.2% (n = 9) and 143.2 +/- 8.1% (n = 6), respectively (P less than 0.01), than was solubilized spontaneously.(ABSTRACT TRUNCATED AT 250 WORDS)
McGowan
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Department of Veterans Affairs Research Service, IA.
D000818:Animals; D056649:Cathepsin G; D002403:Cathepsins / Q000378:metabolism; D002478:Cells, Cultured; D002850:Chromatography, Gel; D005109:Extracellular Matrix / Q000378:metabolism*; D005347:Fibroblasts; D006025:Glycosaminoglycans / Q000378:metabolism; D006801:Humans; D008168:Lung / Q000378:metabolism*; D009131:Muscle, Smooth, Vascular / Q000166:cytology / Q000378:metabolism; D009504:Neutrophils / Q000201:enzymology / Q000378:metabolism*; D010196:Pancreatic Elastase / Q000378:metabolism; D009195:Peroxidase / Q000378:metabolism; D011509:Proteoglycans / Q000378:metabolism*; D051381:Rats; D011919:Rats, Inbred Strains; D012697:Serine Endopeptidases; D012995:Solubility
D006025:Glycosaminoglycans; D011509:Proteoglycans; D009195:Peroxidase; D002403:Cathepsins; D012697:Serine Endopeptidases; C534094:CTSG protein, human; D056649:Cathepsin G; C534097:Ctsg protein, rat; D010196:Pancreatic Elastase