Production of proteolytic enzymes in mast cells, fibroblasts, vascular smooth muscle and endothelial cells cultivated under normoxic or hypoxic conditions.
Matrix metalloproteinases (MMPs) is a family of proteolytic enzymes involved in remodeling of extracellular matrix. Although proteolytic enzymes are produced by many cell types, mast cells seem to be more important than other types in remodeling of pulmonary arteries during hypoxia. Therefore, we tested in vitro production of MMPs and serine proteases in four cell types (mast cells, fibroblasts, vascular smooth muscle cells and endothelial cells) cultivated for 48 h under normoxic or hypoxic (3% O2) conditions. MMP-13 was visualized by immunohistochemistry, MMP-2 and MMP-9 were detected by zymography in cell lysates. Enzymatic activities (MMPs, tryptase and chymase) were estimated in the cultivation media. Hypoxia had a minimal effect on total MMP activity in the cultivation media of all types of cells, but immunofluorescence revealed higher intensity of MMP-13 in the cells exposed to hypoxia except of fibroblasts. Tryptase activity was three times higher and chymase activity twice higher in mast cells cultivated in hypoxia than in those cultured in normoxia. Among all cell types studied here, mast cells are the most abundant source of proteolytic enzymes under normoxic and hypoxic conditions. Moreover, in these cells hypoxia increases the production of both specific serine proteases tryptase and chymase, which can act as MMPs activators.
Maxová
H
H
Department of Pathological Physiology, Second Medical School, Charles University, Prague, Czech Republic. hana.maxova@lfmotol.cuni.cz
Bačáková
L
L
Lisá
V
V
Novotná
J
J
Tomášová
H
H
Vízek
M
M
Herget
J
J
D000818:Animals; D002417:Cattle; D002460:Cell Line; D053818:Chymases / Q000378:metabolism; D042783:Endothelial Cells / Q000166:cytology / Q000201:enzymology*; D005347:Fibroblasts / Q000166:cytology / Q000201:enzymology*; D000860:Hypoxia / Q000378:metabolism*; D008297:Male; D008407:Mast Cells / Q000166:cytology / Q000201:enzymology*; D034801:Mastocytoma; D053509:Matrix Metalloproteinase 13 / Q000378:metabolism; D020778:Matrix Metalloproteinase 2 / Q000378:metabolism; D020780:Matrix Metalloproteinase 9 / Q000378:metabolism; D051379:Mice; D009131:Muscle, Smooth, Vascular / Q000166:cytology; D032389:Myocytes, Smooth Muscle / Q000166:cytology / Q000201:enzymology*; D010100:Oxygen / Q000494:pharmacology; D010447:Peptide Hydrolases / Q000378:metabolism*; D011651:Pulmonary Artery / Q000166:cytology; D051381:Rats; D017208:Rats, Wistar; D053802:Tryptases / Q000378:metabolism
D010447:Peptide Hydrolases; D053818:Chymases; D053802:Tryptases; D053509:Matrix Metalloproteinase 13; D020778:Matrix Metalloproteinase 2; D020780:Matrix Metalloproteinase 9; D010100:Oxygen