title: Moderate inhibition of myocardial matrix metalloproteinase-2 by ilomastat is cardioprotective.
abstract: Pharmacological inhibition of matrix metalloproteinase-2 (MMP-2) is a promising target for acute cardioprotection against ischemia/reperfusion injury. Therefore, here we investigated if the MMP inhibitor ilomastat administered either before ischemia or before reperfusion is able to reduce infarct size via inhibition of MMP-2, the most abundant MMP in the rat heart. Infarct-size limiting effect of ilomastat (0.3-6.0μmol/kg) was tested in an in vivo rat model of myocardial infarction induced by 30min coronary occlusion/120min reperfusion. Ilomastat at 0.75 and 1.5μmol/kg decreased infarct size significantly as compared to the vehicle-treated (dimethyl sulfoxide) group (from 66.1±4.6% to 45.3±7.0% and 46.7±5.5% of area at risk, p<0.0.5, respectively), when administered 5min before the onset of ischemia. Ilomastat at 6.0μmol/kg significantly reduced infarct size from its control value of 65.4±2.5% to 52.8±3.7% of area at risk (p<0.05), when administered 5min before the onset of reperfusion. Area at risk was not significantly affected by ilomastat treatments. To further assess the cytoprotective effect of ilomastat, primary cardiomyocytes isolated from neonatal rats were subjected to 240min simulated ischemia followed by 120min simulated reperfusion in the presence of ilomastat (5nM-5μM). Ilomastat at 500nM and 5μM significantly increased cell viability when compared to vehicle treated group. To assess the in situ MMP-2 inhibitory effect of ilomastat, in separate experiments in situ zymography was performed in cardiomyocytes. The cytoprotective concentration of ilomastat (500nM) showed a moderate (approximately 25%) inhibition of intracellular MMP-2 in ischemic/reperfused cardiomyocytes. In these cells, MMP-2 immunostaining showed a 90% colocalization with the in situ gelatinolytic activity. We conclude that the MMP inhibitor ilomastat reduces infarct size when administered either before the onset of ischemia or before the onset of reperfusion in vivo. Furthermore, this is the first demonstration that a moderate inhibition of intracellular MMP-2 is sufficient to confer cardiocytoprotection.
authors.affiliation: Cardiovascular Research Group, Department of Biochemistry, University of Szeged, Szeged, Hungary; Pharmahungary Group, Szeged, Hungary.
authors.forename: Péter
authors.initials: P
authors.lastname: Bencsik
authors.affiliation: Cardiovascular Research Group, Department of Biochemistry, University of Szeged, Szeged, Hungary.
authors.forename: János
authors.initials: J
authors.lastname: Pálóczi
authors.affiliation: Cardiovascular Research Group, Department of Biochemistry, University of Szeged, Szeged, Hungary.
authors.forename: Gabriella F
authors.initials: GF
authors.lastname: Kocsis
authors.affiliation: Cardiovascular Research Group, Department of Biochemistry, University of Szeged, Szeged, Hungary.
authors.forename: Judit
authors.initials: J
authors.lastname: Pipis
authors.affiliation: Department of Medical Biology, Faculty of Medicine, University of Szeged, Szeged, Hungary.
authors.forename: István
authors.initials: I
authors.lastname: Belecz
authors.affiliation: Cardiovascular Research Group, Department of Biochemistry, University of Szeged, Szeged, Hungary.
authors.forename: Zoltán V
authors.initials: ZV
authors.lastname: Varga
authors.affiliation: Cardiovascular Research Group, Department of Biochemistry, University of Szeged, Szeged, Hungary; Pharmahungary Group, Szeged, Hungary.
authors.forename: Csaba
authors.initials: C
authors.lastname: Csonka
authors.affiliation: Cardiovascular Research Group, Department of Biochemistry, University of Szeged, Szeged, Hungary; Pharmahungary Group, Szeged, Hungary.
authors.forename: Anikó
authors.initials: A
authors.lastname: Görbe
authors.affiliation: Cardiovascular Research Group, Department of Biochemistry, University of Szeged, Szeged, Hungary; Pharmahungary Group, Szeged, Hungary.
authors.forename: Tamás
authors.initials: T
authors.lastname: Csont
authors.affiliation: Pharmahungary Group, Szeged, Hungary; Department of Pharmacology and Pharmacotherapy, Semmelweis University, Budapest, Hungary. Electronic address: peter.ferdinandy@pharmahungary.com.
authors.forename: Péter
authors.initials: P
authors.lastname: Ferdinandy
mesh_terms: D000818:Animals; D002316:Cardiotonic Agents / Q000494:pharmacology / Q000627:therapeutic use*; D002470:Cell Survival / Q000187:drug effects; D004305:Dose-Response Relationship, Drug; D018093:Gelatinases / Q000037:antagonists & inhibitors; D006321:Heart / Q000187:drug effects*; D006877:Hydroxamic Acids; D007211:Indoles / Q000494:pharmacology / Q000627:therapeutic use*; D008297:Male; D061965:Matrix Metalloproteinase Inhibitors / Q000494:pharmacology / Q000627:therapeutic use*; D009203:Myocardial Infarction / Q000188:drug therapy / Q000201:enzymology / Q000473:pathology; D009206:Myocardium / Q000166:cytology / Q000201:enzymology* / Q000473:pathology; D032383:Myocytes, Cardiac / Q000187:drug effects / Q000201:enzymology; D061251:Primary Cell Culture; D051381:Rats; D015427:Reperfusion Injury / Q000188:drug therapy / Q000473:pathology
chemical_list: D002316:Cardiotonic Agents; D006877:Hydroxamic Acids; D007211:Indoles; D061965:Matrix Metalloproteinase Inhibitors; D018093:Gelatinases; C119203:ilomastat
keywords: Cardiomyocyte; Coronary occlusion; Ilomastat; In situ zymography; Ischemia/reperfusion injury; Matrix metalloproteinase