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The angiotensin–calcineurin–NFAT pathway mediates stretch-induced up-regulation of matrix metalloproteinases-2/-9 in atrial myocytes

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Abstract

Aim

During atrial fibrillation, arterial hypertension and systolic or diastolic heart failure, atrial myocytes are exposed to increased baseline stretch. Atrial stretch has been shown to induce cellular hypertrophy and extracellular matrix remodeling (ECM) via angiotensin-II dependent pathways and the matrix metalloproteinases system (MMPs). We hypothesized that atrial myocytes exposed to static stretch may increase their ECM remodeling activity via up-regulation of MMP-2/-9. We then tested the hypothesis that the membrane bound angiotensin-II type 1 (AT1) receptor and the intracellular calcineurin (Cn)-NFAT signaling pathway are potential mediators of stretch-induced MMP alterations, since Cn-NFAT is one important contributor to myocyte hypertrophy.

Methods and results

Neonatal rat atrial myocytes (NRAM) were cultured under conditions of static stretch by 21%. The differential effects of selective AT1 receptor blockade by losartan, Cn blockade by Cyclosporine-A (CsA) or NFAT inhibition by 11R-VIVIT (VIV), were analyzed. Stretch resulted in a significant up-regulation of active-MMP-2/-9 protein amount (active-MMP-2 ng/µg: control 8.95 ± 0.64 vs. stretch 13.11 ± 0.74 / active-MMP-9 ng/µg: control 1.45 ± 0.18 vs. stretch 1.94 ± 0.21, all n = 5) and enzyme activity (MMP-2 in %: control 1 ± 0.0 vs. stretch 1.87 ± 0.25, n = 7) associated with a significant increase of the membrane-type-1-MMP (MT1-MMP) protein expression (MT1-MMP in %: control 1 ± 0.0 vs. stretch 2.17 ± 0.21, n = 8). These observations were accompanied by an activation of the Cn-NFAT pathway (Cn-activity in nmol PO4 release/20 µg protein/30 min: control 0.37 ± 0.08 vs. stretch 0.65 ± 0.09, n = 3 / NFATc1-DNA binding activity in %: control 1 ± 0.0 vs. stretch 1.53 ± 0.17, n = 3). Losartan, CsA or VIV abolished stretch-induced alterations in MMP-2/-9 and MT1-MMP expression and enzyme activity by normalizing the Cn-activity and the DNA binding activity of NFATc1.

Conclusion

Our results present new insights in molecular mechanisms of ECM remodeling activity of atrial myocytes exposed to static stretch. The AT1-Cn-NFAT pathway is a potential mediator of MMP activation.

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Acknowledgments

We thank the entire Institute of Laboratory Animal Science, University Hospital, RWTH Aachen in Germany for the helpful assistance in animal research. This work was supported in part by Deutsche Forschungsgemeinschaft (DFG) grant Lu 869/4-1 and IZKF Biomat. RWTH Aachen, Germany to A.L. and by the Network of Competence Atrial Fibrillation.

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Authors and Affiliations

  1. Dept. of Cardiology, Medical Clinic I, University Hospital Rheinisch Westfälische Technische Hochschule Aachen, Pauwelsstraße 30, 52074, Aachen, Germany

    Erol Saygili MD, Obaida R. Rana, Christian Meyer, Christopher Gemein, Karl Mischke, Tienush Rassaf, Malte Kelm & Patrick Schauerte

  2. Institute for Molecular Cardiovascular Research, University RWTH Aachen, Aachen, Germany

    Michael G. Andrzejewski, Andreas Ludwig & Christian Weber

  3. Dept. of Physiology, University of Maastricht, Maastricht, The Netherlands

    Ulrich Schotten

  4. Institute for Neuropathology, University RWTH Aachen, Aachen, Germany

    Alexander Krüttgen & Joachim Weis

  5. Medical Clinic II, Klinikum Weiden, Weiden, Germany

    Robert H. G. Schwinger

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Correspondence to Erol Saygili MD.

Additional information

Returned for 1. Revision: 15 August 2008 1. Revision received: 17 November 2008

E. Saygili and O. R. Rana contributed equally to this work.

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Saygili, E., Rana, O.R., Meyer, C. et al. The angiotensin–calcineurin–NFAT pathway mediates stretch-induced up-regulation of matrix metalloproteinases-2/-9 in atrial myocytes. Basic Res Cardiol 104, 435–448 (2009). https://doi.org/10.1007/s00395-008-0772-6

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