Role of plasmin and gelatinase in extracellular matrix degradation by cultured rat mesangial cells.
We have examined the ability of mesangial cells (MCs) to degrade extracellular matrix (ECM) using cultured rat MCs grown on thin films of radiolabeled Matrigel. ECM degradation by cultured MCs was observed only in presence of exogenously added plasminogen and was a function of plasminogen concentration (0-50 mU), cell number (0-50,000 cells), and length of incubation (0-72 h). A high positive correlation (r > 0.93) was observed between ECM degradation and plasmin activity in medium, suggesting an important role for plasmin in ECM degradation by cultured MCs. This suggestion was confirmed by ability of plasmin inhibitors, alpha 2-antiplasmin (40 micrograms/ml) and aprotinin (216 kallikrein inhibition units/ml), to inhibit (> 90%) ECM degradation. Inhibitors of cysteine proteinases [trans-epoxysuccinyl-L-leucylamido(4-guanidino)butane, 10 microM] and aspartic proteinases (pepstatin, 5.0 micrograms/ml) had no effect on ECM degradation. However, in presence of plasminogen, inhibitors of matrix metalloproteinases, TIMP-1 (40 micrograms/ml) and o-phenanthroline (100 microM), inhibited ECM degradation -42 +/- 4% and -43 +/- 3% (SE), respectively (n = 8-10). Thus, in addition to plasmin, a matrix metalloproteinase(s) is also involved in ECM degradation by cultured rat MCs. Zymography of culture medium obtained from MCs grown on radiolabeled Matrigel films in absence of plasminogen revealed only two closely migrating bands of gelatinase activity, relative mol wt of approximately 70,000-72,000. MCs grown in absence of plasminogen failed to degrade ECM despite presence of gelatinase in medium, indicating that, in absence of plasmin, gelatinase is present in an inactive form, either as a latent proenzyme or as a gelatinase-inhibitor complex.(ABSTRACT TRUNCATED AT 250 WORDS)
Wong
A P
AP
Department of Biochemistry, Tulane Medical School, New Orleans, Louisiana 70112.
Cortez
S L
SL
Baricos
W H
WH
D000818:Animals; D002478:Cells, Cultured; D003094:Collagen / Q000378:metabolism; D017364:Collagenases / Q000502:physiology*; D004338:Drug Combinations; D005109:Extracellular Matrix / Q000378:metabolism*; D005341:Fibrinolysin / Q000502:physiology*; D005920:Glomerular Mesangium / Q000166:cytology / Q000378:metabolism*; D007797:Laminin / Q000378:metabolism; D020780:Matrix Metalloproteinase 9; D011480:Protease Inhibitors / Q000494:pharmacology; D011509:Proteoglycans / Q000378:metabolism; D051381:Rats
D004338:Drug Combinations; D007797:Laminin; D011480:Protease Inhibitors; D011509:Proteoglycans; C059485:matrigel; D003094:Collagen; D005341:Fibrinolysin; D017364:Collagenases; D020780:Matrix Metalloproteinase 9