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Integrin stimulation-induced hypertrophy in neonatal rat cardiomyocytes is NO-dependent

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Abstract

Prolonged myocardial stretch typically leads to hypertrophy of cardiomyocytes. As integrins are cellular receptors of stretch, we hypothesize that integrin stimulation induces cardiomyocyte hypertrophy. Integrins of neonatal rat cardiomyocytes (NRCMs) were stimulated with a peptide containing the Arg-Gly-Asp (RGD) sequence for 24 h. For comparison, α1-adrenergic stimulation by phenylephrine (PE) for 24 h was applied. Saline-treated NRCMs were used as control. The hypertrophic response was quantified by measuring cell surface area (CSA). Phosphorylation of NO-synthase-1 (NOS1) was assessed by immunocytochemistry. CSA was increased by 38% (IQR 31–44%) with RGD and by 68% (IQR 64–84%) with PE versus control (both P < 0.001). NOS-1 phosphorylation was increased by 61% with RGD and by 21% with PE versus control (both P < 0.01). A general NOS-inhibitor (l-NAME) inhibited RGD-induced hypertrophy completely, but had no significant effect on PE-induced hypertrophy. Administration of NO-donor to NRCMs co-incubated with RGD + l-NAME partly restored hypertrophy (to 62% of the hypertrophic effect of RGD alone), but had no effect if incubated with PE + l-NAME. Ryanodine and BAPTA-AM inhibited RGD-induced hypertrophy completely but not that induced by PE. Integrin stimulation of NRCMs by RGD leads to hypertrophy, likely by activation of NOS-1. Abrogation of RGD-induced hypertrophic response upon NOS-inhibition and rescue of this hypertrophic effect by NO-donor suggest that integrin stimulation-induced hypertrophy of NRCMs is NO-dependent.

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Abbreviations

FAK:

Focal adhesion kinase

nNOS, NOS1:

Neuronal nitric oxide synthase

NO:

Nitric oxide

PBS:

Phosphate-buffered saline

ERK:

Extracellular signal-regulated kinase

MAP kinase:

Mitogen-activated protein kinase

MEK:

Mitogen-activated protein kinase kinase, also indicated by MAPK/ERK kinase

NRCM:

Neonatal rat cardiomyocyte

PE:

Phenylephrine

RGD:

Arg-Gly-Asp-sequence

l-NAME:

Nω-nitro-l-arginine methyl ester

SMTC:

S-methyl-l-thiocitrulline

PKC:

Protein kinase C

BAPTA-AM:

1,2-Bis(2-aminophenoxy)ethane-N,N,N′,N′-tetraacetic acid tetrakis-acetoxymethyl ester

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Acknowledgment

Mrss. W.H. Bax and C.I. Schutte are gratefully acknowledged for expert technical support.

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Authors and Affiliations

  1. Department of Cardiology, Leiden University Medical Center, P.O. Box 9600, 2300 RC, Leiden, The Netherlands

    S. Umar, E. J. M. van der Valk, M. J. Schalij, E. E. van der Wall, D. E. Atsma & A. van der Laarse

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  1. S. Umar
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  2. E. J. M. van der Valk
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  3. M. J. Schalij
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  4. E. E. van der Wall
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Correspondence to A. van der Laarse.

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Umar, S., van der Valk, E.J.M., Schalij, M.J. et al. Integrin stimulation-induced hypertrophy in neonatal rat cardiomyocytes is NO-dependent. Mol Cell Biochem 320, 75–84 (2009). https://doi.org/10.1007/s11010-008-9900-8

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