Elevated Levels of Matrix Metalloproteinase 9 and Tissue Inhibitor of Metalloproteinase 1 during the Acute Phase of Kawasaki Disease

Abstract

Kawasaki disease (KD) is an acute, self-limiting, multisystem vasculitis of unknown etiology affecting infants and young children. Unless treated promptly with high-dose intravenous gamma globulin and aspirin, patients frequently develop coronary aneurysms. Previously, matrix metalloproteinase 9 (MMP-9), which is secreted complexed to tissue inhibitor of metalloproteinase 1 (TIMP-1), has been implicated in abdominal aortic aneurysm formation. Since the clinical and pathological features of KD include inflammation and weakening of blood vessels, we analyzed acute-and convalescent-phase paired plasma or serum samples from 31 KD patients, 7 patients who did not completely meet the criteria for KD, and 26 non-KD controls (9 febrile and 17 afebrile patients) for pro-MMP-9 (92 kDa) enzyme activity by gelatin zymography and for active MMP-9 (83 kDa), pro-MMP-9, and TIMP-1 protein levels by enzyme-linked immunosorbent assay. Statistical analysis was performed by using Student t tests, linear regression, and the Wilcoxon rank-sum test. Markedly elevated pro-MMP-9 enzymatic activity, pro-MMP-9 protein levels, and TIMP-1 protein levels were found during the acute phase of illness in patients with clinically established KD and in patients who were suspected of having KD but did not meet all of the criteria. There was no significant difference in active MMP-9 levels. Furthermore, pro-MMP-9 and TIMP-1 protein levels were significantly elevated among KD patients, compared to those of febrile and afebrile non-KD controls. The significantly elevated pro-MMP-9 enzyme and protein levels during the acute phase of KD may reflect vascular remodeling or an inflammatory response to a microbial agent, suggesting a pathophysiological role for MMP-9 in coronary aneurysm formation.

Matrix metalloproteinase 9 activity leads to elastin breakdown in an animal model of Kawasaki disease

“…But despite the fact that MMP-9 levels were elevated in affected hearts in our murine model of KD, there was no correlation between circulating MMP-9 levels and enzymatic activity in affected coronary arteries (25). This is in accordance with data from cases of fatal KD, which show MMP-9 expression specifically localized to the site of coronary artery lesions (26), and the conflicting reports regarding circulating MMP-9 levels in affected children (22)(23)(24)(25). Degradation of elastin is one of the hallmarks of aneurysm formation.…”

“…MMP-9 may have a role in coronary artery aneurysm formation in KD. Despite conflicting data on serum levels of MMP-9 during the acute phase of KD (22)(23)(24)(25), local MMP-9 expression at the site of coronary artery aneurysms in children with KD has been documented (26). Circulating levels of molecules known to act locally may not be useful biomarkers of disease; this is especially relevant to enzymatic activity, which is tightly regulated at multiple levels including the local tissue environment (25).…”

IL-10 Is a Negative Regulatory Factor of CAWS-Vasculitis in CBA/J Mice as Assessed by Comparison with Bruton’s Tyrosine Kinase-Deficient CBA/N Mice

“…3). It was reported that there was no correlation between blood TIMP and MMP production levels and disease severity in multiple patients with Kawasaki disease, and an imbalance of these levels contributed to the development of vasculitis (24,25). It was also suggested that the imbalance of TIMP and MMP is a factor contributing to the development of vasculitis.…”

Curcumin blocks CCL2-induced adhesion, motility and invasion, in part, through down-regulation of CCL2 expression and proteolytic activity

“…The proteolytic enzyme activity of pro-MMP-9 was measured by gelatin zymography (18). Briefly, protein concentrations were determined with the Bio-Rad DC (Bio-Rad Laboratories; Hercules, CA) protein assay kit from conditioned media and cell-lysate of PC-3 cells cultured under growing conditions as described previously (18).…”

“…The proteolytic enzyme activity of pro-MMP-9 was measured by gelatin zymography (18). Briefly, protein concentrations were determined with the Bio-Rad DC (Bio-Rad Laboratories; Hercules, CA) protein assay kit from conditioned media and cell-lysate of PC-3 cells cultured under growing conditions as described previously (18). Total protein (15 μg) was mixed with non-denaturing buffer (125 mM Tris-HCl, 4% SDS, 20% glycerol, 0.004% bromophenol blue) and the protein samples were subjected to electrophoresis through a 10% Zymogram Ready Gel (Bio-Rad Laboratories).…”