Preconditioning decreases ischemia/reperfusion-induced release and activation of matrix metalloproteinase-2

doi:10.1016/S0006-291X(02)02019-3

Biochemical and Biophysical Research Communications

Volume 296, Issue 4, 30 August 2002, Pages 937-941

https://doi.org/10.1016/S0006-291X(02)02019-3 Get rights and content

Abstract

Release and activation of matrix metalloproteinases (MMPs) significantly contribute to myocardial stunning injury immediately after ischemia and reperfusion, however, their role in preconditioning remains unknown. We therefore examined the effects of preconditioning and subsequent ischemia/reperfusion on MMP activity in isolated rat hearts. Hearts were subjected to a preconditioning protocol (three consecutive 5-min periods of global ischemia interspersed with 5 min of reperfusion) followed by 30 min ischemia and 5 min reperfusion. To measure MMP release, coronary effluent was collected: (a) during aerobic perfusion, (b) in reperfusion following each preconditioning ischemia, and (c) during the final reperfusion following test ischemia. MMP-2 activities could be detected by gelatin zymography in the ventricles and coronary effluent samples from the perfused hearts. The levels of MMP-2 activity in the effluent were markedly increased in effluent following test ischemia from control hearts without preconditioning. This was accompanied by a decrease in corresponding tissue MMP activities. Preconditioning significantly decreased the MMP-2 activity in the coronary effluent following test ischemia/reperfusion and preserved the MMP-2 protein content and activity in the myocardium. Our results demonstrate that classic preconditioning inhibits ischemia/reperfusion induced release and activation of MMP-2. These results suggest that preconditioning may exert part of its cardioprotective effects through the reduction of MMP-2 release.

Section snippets

Materials and methods

The investigation conforms to the Guide for the Care and Use of Laboratory Animals published by the National Research Council (revised 1996) and was approved by local ethics committees.

Isolated heart preparation. Hearts from male Sprague–Dawley rats (250–300 g, anesthetized with 100 mg/kg pentobarbital, and given 500 U/kg heparin) were isolated and briefly rinsed by immersion in ice-cold Krebs–Henseleit buffer. Spontaneously beating hearts were retrogradely perfused according to the Langendorff

Results

Gelatinolytic activities could be detected by zymography at 75, 72, and 62 kDa in the ventricles and all coronary effluents of perfused rat hearts. The 72- and 62-kDa forms were identified as MMP-2 by comparison to the HT-1080 standard. The 75-kDa band has been identified as a rat/murine specific glycosylated form of MMP-2 (personal communication, Chris Overall, University of British Columbia). MMP-9 gelatinolytic activity could be detected in neither the effluent nor the ventricles. All

Discussion

This is the first demonstration that ischemic-preconditioning inhibits the release of MMP-2 induced by test ischemia/reperfusion.

MMP-2 is an important species of metalloproteinase which can be found in normal cardiac myocytes [14], cardiac fibroblasts [15], and endocardial cells [16]. MMPs are synthesized as pro-enzymes and are usually activated by proteolytic cleavage of an inhibitory pro-peptide domain. However, it has also been shown that peroxynitrite can activate this enzyme by oxidizing

Acknowledgements

This work was supported by grants from the North Atlantic Treaty Organization Cooperative Linkage grant (NATO, LST.CLG.976650), Hungarian Scientific Research Fund (OTKA-T029843), Hungarian Ministry of Education (FKFP-0340/2000, FKFP-0057/2001), the Hungarian National R&D Program (NFKP-1/040), and the Canadian Institutes of Health Research (MT-14741). P.F. is a Bolyai fellow of the Hungarian Academy of Sciences. R.S. is a senior scholar and T.C. is a fellow of the Alberta Heritage Foundation for

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It is well-established that reperfusion of the ischemic myocardium increases MMP-2 activity in the coronary effluent in vitro [4] and in plasma in vivo within minutes [67]. In addition, our previous studies have consistently demonstrated that the same IPC protocol used in the present experiment prevented the early release of MMP-2 from the isolated heart, preserved cardiac mechanical function and limited infarct size [68], indicating that inhibition of early MMP-2 activation is an effector mechanism in IPC-evoked cardioprotection. However, to our best knowledge, there is no available data on the effect of cardiac IPC in vivo on the time-course of plasma MMP-2 activity.
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The cardioprotective cellular mechanism in which MMP-2 inhibition might be involved is not known and has not been investigated in the present study. Although endogenous cardioprotection by early and late ischemic preconditioning as well as postconditioning involve an MMP-2 inhibition-dependent mechanism [3,5,6,32] the exact mechanism by which MMP inhibition results in cardioprotection is not known. Bell et al. reported that ilomastat protects the heart against reperfusion injury independently from the well-known cardioprotective Reperfusion Injury Salvage Kinase/mitochondrial permeability transition pore opening pathways [15].
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Preconditioning decreases ischemia/reperfusion-induced release and activation of matrix metalloproteinase-2

Abstract

Section snippets

Materials and methods

Results

Discussion

Acknowledgements

Biochem. Biophys. Res. Commun.

Anal. Biochem.

J. Biol. Chem.

Release of gelatinase A during platelet activation mediates aggregation

Nature

Vascular matrix metalloproteinase-2 cleaves big endothelin-1 yielding a novel vasoconstrictor

Circ. Res.

Vascular matrix metalloproteinase-2-dependent cleavage of calcitonin gene-related peptide promotes vasoconstriction

Circ. Res.

Inflammation dampened by gelatinase A cleavage of monocyte chemoattractant protein-3

Science

Matrix metalloproteinase-2 contributes to ischemia–reperfusion injury in the heart

Circulation

Peroxynitrite-induced myocardial injury is mediated through matrix metalloproteinase-2

Cardiovasc. Res.