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The effect of chronic nitric oxide synthases inhibition on regulatory proteins in rat hearts

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Abstract

The aim of the present study was to characterize the effects of chronic nitric oxide synthase (NOS) inhibition on the alterations of regulatory myocardial proteins of intracellular signaling pathways (mitogen-activated protein kinase (MAPK) and Akt kinase cascades) and matrix metalloproteinases (MMP). Chronic NO deficiency (NOD) was induced by NG-nitro-L-arginine methyl ester (L-NAME, 40mg/kg/day, 4 weeks). Protein levels and activation of protein kinases were determined using specific antibodies, activities of MMP were analyzed by zymography in gels containing gelatin as a substrate. The development of NOD was associated with decreased activation of endothelial NOS (eNOS) and down-regulation of protein level of inducible NOS (iNOS). Investigation of kinase pathways revealed that the activation of extracellular signal-regulated kinases (ERK) and the levels of upstream activators of ERK (aFGF, H-Ras) were decreased after L-NAME treatment. Western blot analysis revealed that chronic application of L-NAME also decreased the activation of Akt kinase as compared with control hearts. Study of MMPs showed that in L-NAME-treated rat hearts activities of tissue MMP-2 were decreased. It is concluded that development of NOD resulted in inhibition of ERK and Akt kinase pathways and these changes suggest the involvement of these cascades in responses of myocardium to NOD. The results also point to the possible relationship between ERK and Akt kinase pathways and activation of eNOS and/or MMP-2.

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Acknowledgment

Supported by VEGA SR grants No. 2/6170/26, 2/5110/27, 2/6148/27, APVV 51-027404, SP51/028 09 00/028 09 01.

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Authors and Affiliations

  1. Institute for Heart Research, Centre of Excellence for Cardiovascular Research, Slovak Academy of Sciences, Dúbravská cesta 9, P.O. Box 104, 840 05, Bratislava, Slovak Republic

    Anna Špániková, Petra Šimončíková, Táňa Ravingerová & Miroslav Barančík

  2. Institute of Molecular Physiology and Genetics, Centre of Excellence for Cardiovascular Research, Slovak Academy of Sciences, Bratislava, Slovak Republic

    Anna Špániková

  3. Institute of Normal and Pathological Physiology, Centre of Excellence for Cardiovascular Research, Slovak Academy of Sciences, Bratislava, Slovak Republic

    Oľga Pecháňová

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  1. Anna Špániková
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  2. Petra Šimončíková
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  3. Táňa Ravingerová
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  4. Oľga Pecháňová
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  5. Miroslav Barančík
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Correspondence to Miroslav Barančík.

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Anna Špániková and Petra Šimončíková have contributed equally to the study.

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Špániková, A., Šimončíková, P., Ravingerová, T. et al. The effect of chronic nitric oxide synthases inhibition on regulatory proteins in rat hearts. Mol Cell Biochem 312, 113–120 (2008). https://doi.org/10.1007/s11010-008-9726-4

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  • DOI: https://doi.org/10.1007/s11010-008-9726-4

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