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Nitric oxide/cGMP signalling mediates the cardioprotective action of adrenomedullin in reperfused myocardium

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Abstract

We demonstrated previously that adrenomedullin (AM), when given during early reperfusion, limited infarct size in rat heart. The present study was undertaken to provide direct evidence of the NO-dependency of AM’s cardioprotective action by assessing NO biosynthesis and involvement of the soluble guanylyl cyclase (sGC) pathway. Perfused hearts from male CD-1 mice were subjected to 30-min left coronary occlusion and 60-min reperfusion. Infarct size was determined by tetrazolium staining. AM 10 nM was administered from 20 min after coronary occlusion until 10 min after reperfusion. Coronary effluent was analysed for NO2 − and NO3 −, and myocardial samples were analysed for NO2 −, NO3 −, nitroso-adducts and cGMP concentration. To examine the role of NO/sGC signalling in the infarct-limiting action of AM, further hearts received the sGC inhibitor ODQ 2 μM. AM treatment stimulated NO synthesis, indicated by increased NO2 − efflux in coronary effluent throughout reperfusion (summarised as area under curve, AM 29.2 ± 3.9 vs. control 14.4 ± 2.8 μmol min2 mL−1, P < 0.05). AM limited infarct size (35.4 ± 2.7 vs. 12.2 ± 2.3%, P < 0.01), associated with a 2.45-fold increase (P < 0.05) in myocardial cGMP concentration at 10 min after reperfusion. ODQ abolished the infarct size-limiting effect of AM (28.9 ± 4.3%). These data provide the first evidence that AM increases NO bioavailability in intact murine myocardium and confirm that the NO/sGC/cGMP pathway is central to the cytoprotective action of AM against ischaemia–reperfusion injury.

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Abbreviations

AM:

Adrenomedullin

cGMP:

Guanosine-3′, 5′-cyclic monophosphate

mPTP:

Mitochondrial permeability transition pore

NO:

Nitric oxide

NO2 − :

Nitrite

NO3 − :

Nitrate

NOS:

Nitric oxide synthase

ODQ:

1H[1,2,4]oxodiazole-[4,3-a]-quinoxalin-1-one

PI3K:

Phosphoinositol-3-kinase

PKG:

Protein kinase G

RXNOs:

Total nitros(yl)ated chemical species

sGC:

Soluble guanylyl cyclase

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Acknowledgments

GFB and SAH acknowledge support for this work provided by the British Heart Foundation (PG/03/030). The work was also sponsored in part by the German DFG (RA969/4-1 to TR, and GRK1089/TP3 to TR, MT and CD).

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Authors and Affiliations

  1. Division of Pharmacology, Welsh School of Pharmacy, Cardiff University, King Edward VII Avenue, Cardiff, CF10 3NB, UK

    Shabaz A. Hamid & Gary F. Baxter

  2. Division of Cardiology, Pulmonology, and Angiology, University Hospital Aachen, Aachen, Germany

    Matthias Totzeck, Christina Drexhage & Tienush Rassaf

  3. The Royal Veterinary College, University of London, London, UK

    Shabaz A. Hamid, Iain Thompson & Robert C. Fowkes

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  1. Shabaz A. Hamid
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  2. Matthias Totzeck
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  3. Christina Drexhage
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  4. Iain Thompson
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  5. Robert C. Fowkes
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  6. Tienush Rassaf
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  7. Gary F. Baxter
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Correspondence to Gary F. Baxter.

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Hamid, S.A., Totzeck, M., Drexhage, C. et al. Nitric oxide/cGMP signalling mediates the cardioprotective action of adrenomedullin in reperfused myocardium. Basic Res Cardiol 105, 257–266 (2010). https://doi.org/10.1007/s00395-009-0058-7

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  • DOI: https://doi.org/10.1007/s00395-009-0058-7

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