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.
- MeSH
- arteria pulmonalis cytologie MeSH
- buněčné linie MeSH
- chymasy metabolismus MeSH
- endoteliální buňky cytologie enzymologie MeSH
- fibroblasty cytologie enzymologie MeSH
- financování organizované MeSH
- hypoxie metabolismus MeSH
- krysa rodu rattus MeSH
- kyslík farmakologie MeSH
- mastocytom MeSH
- mastocyty cytologie enzymologie MeSH
- matrixová metaloproteinasa 13 metabolismus MeSH
- matrixová metaloproteinasa 2 metabolismus MeSH
- matrixová metaloproteinasa 9 metabolismus MeSH
- myocyty hladké svaloviny cytologie enzymologie MeSH
- myši MeSH
- potkani Wistar MeSH
- proteasy metabolismus MeSH
- skot MeSH
- svaly hladké cévní cytologie MeSH
- tryptasy metabolismus MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- mužské pohlaví MeSH
- myši MeSH
- skot MeSH
- zvířata MeSH
haracterized by fibrotization and muscularization of the walls of peripheral pulmonary arteries. This vessel remodeling is accompanied by an increase in the amount of lung mast cells (LMC) and the presence of small collagen cleavage products in the vessel walls. We hypothesize that hypoxia activates LMC, which release matrix metalloproteinases (MMPs) cleaving collagen and starting increased turnover of connective tissue proteins. This study was designed to determine whether in vitro hypoxia stimulates production of MMPs in rat LMC and increases their collagenolytic activity. The LMC were separated on the Percoll gradient and then were divided into two groups and cultivated for 24 h in 21 % O2+ 5 % CO2 or in 10 % O2 + 5 % CO2. Presence of the rat interstitial tissue collagenase (MMP-13) in LMC was visualized by immunohistological staining and confirmed by Western blot analysis. Total MMPs activity and tryptase activity were measured in both cultivation media and cellular extracts. Exposure to hypoxia in vitro increased the amount of cells positively labeled by anti-MMP-13 antibody as well as activities of all measured enzymes. The results therefore support the concept that LMC are an important source of increased collagenolytic activity in chronic hypoxia.
- Klíčová slova
- Hypoxia in vitro, Rat lung mast cells, Tissue metalloproteinases, Tryptase, Remodeling of pulmonary vessels,
- MeSH
- financování organizované MeSH
- hypoxie buňky MeSH
- imunohistochemie MeSH
- kolagen metabolismus MeSH
- krysa rodu rattus MeSH
- kultivační média metabolismus MeSH
- kultivované buňky MeSH
- mastocyty enzymologie MeSH
- matrixová metaloproteinasa 13 metabolismus MeSH
- matrixová metaloproteinasa 2 metabolismus MeSH
- matrixová metaloproteinasa 9 metabolismus MeSH
- matrixové metaloproteinasy metabolismus MeSH
- plíce cytologie enzymologie MeSH
- potkani Wistar MeSH
- separace buněk MeSH
- tryptasy metabolismus MeSH
- upregulace MeSH
- western blotting MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- mužské pohlaví MeSH
- zvířata MeSH
BACKGROUND: Secondary brain injury contributes to poor outcome for patients sustaining brain trauma. Matrix metalloproteinase-9 (MMP-9) is a potential marker, as well as effector of secondary brain injury. This enzyme degrades components of extracellular matrix, and thus it can contribute to blood-brain barrier disruption.
- MeSH
- časové faktory MeSH
- dospělí MeSH
- ELISA metody MeSH
- financování organizované MeSH
- Glasgowská stupnice kómat MeSH
- lidé středního věku MeSH
- lidé MeSH
- matrixová metaloproteinasa 9 krev MeSH
- mladý dospělý MeSH
- nelineární dynamika MeSH
- pilotní projekty MeSH
- poranění mozku krev MeSH
- senioři nad 80 let MeSH
- senioři MeSH
- Check Tag
- dospělí MeSH
- lidé středního věku MeSH
- lidé MeSH
- mladý dospělý MeSH
- mužské pohlaví MeSH
- senioři nad 80 let MeSH
- senioři MeSH
- ženské pohlaví MeSH
- MeSH
- dítě MeSH
- exokrinní pankreatická insuficience diagnóza MeSH
- feces enzymologie MeSH
- finanční podpora výzkumu jako téma MeSH
- lidé MeSH
- pankreatická elastasa analýza diagnostické užití MeSH
- Check Tag
- dítě MeSH
- lidé MeSH
- Publikační typ
- kongresy MeSH
