Matrix metalloproteinase and elastase activities in LPS-induced acute lung injury in guinea pigs.
Matrix metalloproteinases (MMPs) and elastase are proteolytic enzymes specifically directed against extracellular matrix (ECM) components. They are secreted by inflammatory cells and may consequently contribute to the lesions of the ECM observed during acute pulmonary edema. We therefore evaluated the MMP and elastase activities, which are secreted by cultured alveolar macrophages (AMACs) and polymorphonuclear neutrophils (PMNs) and present in the bronchoalveolar lavage (BAL) fluid in a guinea pig model of acute lung injury induced by intratracheal instillation of lipopolysaccharide (LPS). The control group was given 0.9% NaCl. 24 h after instillation, a BAL was performed, the BAL fluid was separated from the cells by centrifugation, and AMACs and PMNs were separately cultured for 24 h. In BAL fluid from LPS-treated guinea pigs, we found 1) an increase in free gelatinase activity, tested on [3H]gelatin (0.7 +/- 0.2 micrograms.200 microliters BAL fluid-1.48 h-1 vs. 0.2 +/- 0.1 in controls, P < 0.05), and 2) increased total gelatinase activities, as assessed by zymography. The molecular masses of the major gelatinase species found in BAL fluid by zymography were 92 and 68 kDa. The 92-kDa gelatinase was secreted by both AMACs and PMNs, as demonstrated by zymography of their respective culture media. When tested on [3H]elastin, the elastase activity of BAL fluid of LPS-treated animals exhibited no increase, but when tested on a synthetic peptidic substrate [N-succinyl-(L-alanine)3-p-nitro anilide (SLAPN)], increased elastase-like activity was observed (from 17 +/- 4 nmol of SLAPN.200 microliters BAL fluid-1.24 h-1 in control group to 34 +/- 8 in LPS group, P < 0.05). This increase was attributable to the activity of a metalloendopeptidase that was inhibited by the metal chelator EDTA but not by the specific tissue inhibitor of MMPs.
D'Ortho
M P
MP
Institut National de la Santé et de la Recherche Médicale U 296, Faculté de Médecine, Créteil, France.
Jarreau
P H
PH
Delacourt
C
C
Macquin-Mavier
I
I
Levame
M
M
Pezet
S
S
Harf
A
A
Lafuma
C
C
D000818:Animals; D001992:Bronchoalveolar Lavage Fluid / Q000737:chemistry / Q000166:cytology; D004549:Elastin / Q000378:metabolism; D005109:Extracellular Matrix / Q000201:enzymology*; D006168:Guinea Pigs; D008070:Lipopolysaccharides / Q000494:pharmacology*; D008168:Lung / Q000187:drug effects* / Q000201:enzymology* / Q000473:pathology; D008297:Male; D008666:Metalloendopeptidases / Q000378:metabolism*; D010196:Pancreatic Elastase / Q000378:metabolism*; D010447:Peptide Hydrolases / Q000378:metabolism
D008070:Lipopolysaccharides; D004549:Elastin; D010447:Peptide Hydrolases; D010196:Pancreatic Elastase; D008666:Metalloendopeptidases