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Inhibiting matrix metalloproteinase-2 reduces protein release into coronary effluent from isolated rat hearts during ischemia-reperfusion

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Abstract

Background

Previous studies have shown that the disruption of the coronary endothelium and the increase in its permeability during ischemia-reperfusion (I/R), are linked to matrix metalloproteinase-2 (MMP-2) activity. Studies from our group have shown that during I/R, activity of MMP-2 in the coronary effluent increases and this increase is associated with cardiac dysfunction, which in turn, can be prevented by MMP inhibitors. Therefore, we hypothesize that inhibiting MMPs reduces the MMP-2 dependent disruption of the coronary endothelium and subsequent protein release during I/R.

Methods

Isolated rat hearts were perfused in the Langendorff mode at a constant pressure and subjected to 15, 20 or 30 min no-flow ischemia followed by 30 min of reperfusion. The MMP inhibitors, o-phenanthroline (Phen, 100 µM) or doxycycline (Doxy, 30 µM) an inhibitors of MMPs, were added to the perfusion solution 10 min before ischemia and for the first 10 min of reperfusion. The coronary effluents were collected during perfusion for protein analysis. Creatine kinase was measured as an index of cellular damage. Endothelial integrity was assessed by measuring coronary flow and by measuring the levels of serotransferrin and interstitial albumin in the coronary effluent. Additionally, damage to the endothelium was assessed histologically by light microscopy analysis of the cellular structure of the myocardium. MMP-2 activity was measured by zymography in hearts subjected to 15, 20 and 30 min of ischemia without reperfusion.

Results

MMP-2 activity was increased in heart tissue at the end of ischemia and was correlated with duration of ischemia. The post-ischemia decrease in coronary flow, and the increase in the release of serotransferrin and albumin were attenuated by Phen. Edema (another indirect marker of endothelial damage) was observed in I/R heart and the edema was abolished in I/R heart treated with MMP inhibitors.

Conclusion

MMP inhibition not only reduces cardiac mechanical dysfunction but also reduces endothelial damage resulting from cardiac I/R injury.

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Article Open access 06 August 2014

Heleen M Oudemans-van Straaten, Angelique ME Spoelstra-de Man & Monique C de Waard

Abbreviations

I/R:

Ischemia-reperfusion

MMPs:

Matrix metalloproteinases

Phen:

o-Phenanthroline

Doxy:

Doxycycline

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Acknowledgments

We thank Dr. J. S Richardson for help with manuscript preparation. This work was supported by grants from Canadian Institutes of Health Research and the Heart and Stroke Foundation of Canada.

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Author notes

  1. Justyna Fert-Bober

    Present address: Dept. of Pharmacology, College of Medicine, University of Saskatchewan, 107 Wiggins Road, Saskatoon, SK, S7N 5E5, Canada

Authors and Affiliations

  1. Dept. of Pharmacology, College of Medicine, University of Saskatchewan, 107 Wiggins Road, Saskatoon, SK, S7N 5E5, Canada

    Jolanta Sawicka & Grzegorz Sawicki

  2. Dept. of Clinical Chemistry, Wroclaw Medical University, Wroclaw, Poland

    Justyna Fert-Bober

  3. Dept. of Pediatrics and Pharmacology, University of Alberta, Edmonton, AB, Canada

    Hernando Leon & Richard Schulz

  4. Dept. of Medicine, Division of Cardiology, University of Saskatchewan, Saskatoon, SK, Canada

    Rashpal S. Basran

  5. Dept. of Anatomy and Cell Biology, University of Saskatchewan, Saskatoon, SK, Canada

    Richard M. Devon

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  1. Justyna Fert-Bober
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Corresponding author

Correspondence to Grzegorz Sawicki.

Additional information

Returned for 1. Revision: 13 December 2007 1. Revision received: 17 March 2008

Returned for 2. Revision: 3 April 2008 2. Revision received: 8 April 2008

J. Fert-Bober and H. Leon contributed equally to this work.

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Fert-Bober, J., Leon, H., Sawicka, J. et al. Inhibiting matrix metalloproteinase-2 reduces protein release into coronary effluent from isolated rat hearts during ischemia-reperfusion. Basic Res Cardiol 103, 431–443 (2008). https://doi.org/10.1007/s00395-008-0727-y

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