Inhibition of matrix metalloproteinases prevents peroxynitrite-induced contractile dysfunction in the isolated cardiac myocyte

H. León, I. Baczkó, G. Sawicki, P. E. Light, R. Schulz

Research output: Contribution to journalArticle

30 Citations (Scopus)

Abstract

Background and purpose: The potent oxidant peroxynitrite (ONOO -) induces mechanical dysfunction in the intact heart in part through activation of matrix metalloproteinase-2 (MMP-2). This effect may be independent of the proteolytic actions of MMPs on extracellular matrix proteins. The purpose of this study was to examine the effects of ONOO - on contractile function at the level of the single cardiac myocyte and whether this includes the action of MMPs. Experimental approach: Freshly isolated ventricular myocytes from adult rats were superfused with Krebs-Henseleit buffer at 21°C and paced at 0.5 Hz. Contractility was measured using a video edge-detector. ONOO - or decomposed ONOO - (vehicle control) were co-infused over 40 min to evaluate the contraction cease time (CCT). The effects of ONOO - on intracellular [Ca 2+] were determined in myocytes loaded with calcium green-1 AM. MMP-2 activity was measured by gelatin zymography. Key results: ONOO - (30-600 μM) caused a concentration-dependent reduction in CCT. Myocytes subjected to 300 μM ONOO - had a shorter CCT than decomposed ONOO - (14.9+1.5 vs 32.2+3.5 min, n=7-8; P-. ONOO - caused shorter calcium transient cease time and significant alterations in intracellular [Ca 2+] homoeostasis which were partially prevented by doxycycline. Conclusions and implications: This is the first demonstration that inhibition of MMPs protects the cardiac myocyte from ONOO --induced contractile failure via an action unrelated to proteolysis of extracellular matrix proteins.

Original languageEnglish
Pages (from-to)676-683
Number of pages8
JournalBritish Journal of Pharmacology
Volume153
Issue number4
DOIs
Publication statusPublished - Feb 2008

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Peroxynitrous Acid
Matrix Metalloproteinases
Cardiac Myocytes
Muscle Cells
Extracellular Matrix Proteins
Matrix Metalloproteinase 2
Doxycycline
Gelatin
Oxidants
Proteolysis
Homeostasis
Calcium

Keywords

  • Calcium homoeostasis
  • Cardiac myocyte
  • Doxycycline
  • Matrix metalloproteinases
  • Peroxynitrite

ASJC Scopus subject areas

  • Pharmacology

Cite this

Inhibition of matrix metalloproteinases prevents peroxynitrite-induced contractile dysfunction in the isolated cardiac myocyte. / León, H.; Baczkó, I.; Sawicki, G.; Light, P. E.; Schulz, R.

In: British Journal of Pharmacology, Vol. 153, No. 4, 02.2008, p. 676-683.

Research output: Contribution to journalArticle

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AB - Background and purpose: The potent oxidant peroxynitrite (ONOO -) induces mechanical dysfunction in the intact heart in part through activation of matrix metalloproteinase-2 (MMP-2). This effect may be independent of the proteolytic actions of MMPs on extracellular matrix proteins. The purpose of this study was to examine the effects of ONOO - on contractile function at the level of the single cardiac myocyte and whether this includes the action of MMPs. Experimental approach: Freshly isolated ventricular myocytes from adult rats were superfused with Krebs-Henseleit buffer at 21°C and paced at 0.5 Hz. Contractility was measured using a video edge-detector. ONOO - or decomposed ONOO - (vehicle control) were co-infused over 40 min to evaluate the contraction cease time (CCT). The effects of ONOO - on intracellular [Ca 2+] were determined in myocytes loaded with calcium green-1 AM. MMP-2 activity was measured by gelatin zymography. Key results: ONOO - (30-600 μM) caused a concentration-dependent reduction in CCT. Myocytes subjected to 300 μM ONOO - had a shorter CCT than decomposed ONOO - (14.9+1.5 vs 32.2+3.5 min, n=7-8; P-. ONOO - caused shorter calcium transient cease time and significant alterations in intracellular [Ca 2+] homoeostasis which were partially prevented by doxycycline. Conclusions and implications: This is the first demonstration that inhibition of MMPs protects the cardiac myocyte from ONOO --induced contractile failure via an action unrelated to proteolysis of extracellular matrix proteins.

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