Matrix-dependent mechanism of neutrophil-mediated release and activation of matrix metalloproteinase 9 in myocardial ischemia/reperfusion.
BACKGROUND
A key component of reperfusion of myocardial infarction is an immediate inflammatory response, which enhances tissue repair. Matrix turnover is crucial to tissue repair, and matrix metalloproteinases (MMPs) are key enzymes involved in matrix degradation. The hypothesis tested is that one inflammation-based effector of tissue repair is the secretion and activation of MMP-9 by infiltrating neutrophils.


RESULTS
Cardiac lymph and tissue were assayed for atent and active MMP-2 and MMP-9 by zymography and immunochemistry. Dual-labeling immunofluorescence determined the cellular source of MMP-9 protein. Isolated canine neutrophils were incubated with preischemic and postischemic cardiac lymph in the presence and absence of collagen-fibronectin pads, and the supernatants were assayed for latent and active MMP-9. MMP-9 increased during the first hours of reperfusion in both lymph supernatants and myocardial extracts, and this increase was of neutrophil origin. MMP-9 in the cardiac lymph remained latent but was activatable. In contrast, MMP-9 in the myocardium was in both latent and active forms. In situ zymography demonstrated that activated MMP-9 surrounded the infiltrated neutrophils. When postischemic cardiac lymph was incubated with neutrophils in vitro, MMP-9 secretion and activation occurred only in the presence of a collagen-fibronectin substrate; preischemic cardiac lymph did not induce significant secretion or activation.


CONCLUSIONS
Infiltrating neutrophils are an early source of MMP-9 after reperfusion, and a portion of MMP-9 in the myocardium is active. Infiltrating neutrophils may localize MMP-9 activation by secreting MMP-9 and as a source of activating proteases.
Lindsey
M
M
Section of Cardiovascular Sciences, DeBakey Heart Center, Department of Medicine, Methodist Hospital, Houston, Texas, USA.
Wedin
K
K
Brown
M D
MD
Keller
C
C
Evans
A J
AJ
Smolen
J
J
Burns
A R
AR
Rossen
R D
RD
Michael
L
L
Entman
M
M
D000818:Animals; D002478:Cells, Cultured; D002634:Chemotaxis, Leukocyte; D003094:Collagen; D003594:Cytoplasmic Granules / Q000201:enzymology; D004285:Dogs; D004789:Enzyme Activation; D005109:Extracellular Matrix / Q000502:physiology*; D005260:Female; D005353:Fibronectins; D007249:Inflammation; D008196:Lymph / Q000166:cytology; D008297:Male; D020780:Matrix Metalloproteinase 9 / Q000378:metabolism / Q000502:physiology*; D017202:Myocardial Ischemia / Q000201:enzymology* / Q000473:pathology; D015428:Myocardial Reperfusion Injury / Q000201:enzymology* / Q000473:pathology; D009504:Neutrophils / Q000201:enzymology / Q000378:metabolism*
D005353:Fibronectins; D003094:Collagen; D020780:Matrix Metalloproteinase 9