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Nitric Oxide is Involved in the Down-regulation of SOST Expression Induced by Mechanical Loading

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Abstract

Mechanical stimulation reduces sclerostin expression in rodents. However, few data are available about the effect of physical stimuli in human systems. Recently we observed that the demethylating agent AzadC induces SOST expression in bone cells. This allowed us in this study to explore the effect of mechanical loading on SOST expression by subjecting AzadC-treated human bone cells to pulsating fluid flow (PFF). PFF significantly decreased the AzadC-induced expression of SOST. This effect persisted for at least 24 h, and in fact SOST expression was lower at 24 h after PFF treatment than at 1 h after PFF treatment (PFF/static ratio 0.47 ± 0.04 vs. 0.63 ± 0.03 respectively, p = 0.03). The PFF-induced decrease in SOST expression was not due to a change in the methylation profile of the SOST promoter. However, PFF stimulated nitric oxide (NO) synthesis, which appeared essential for the PFF effect on SOST expression. In fact, the NO synthase inhibitor 1400 W prevented the effect of PFF on SOST expression. Moreover, the NO-donor SNAP decreased SOST mRNA in bone organ cultures. The conditioned medium (CM) of cells subjected to PFF induced a 38 ± 4 % decrease in SOST expression (p = 0.03) in static cultures and diminished the transcriptional activity of reporter vectors with the cloned SOST promoter (Static-CM: 1.47 ± 0.10 vs. PFF-CM: 0.78 ± 0.09, p = 0.02). This is consistent with a PFF-induced secretion of factors that modulate SOST. Our results suggest that NO and other soluble factors are involved in the inhibition of SOST expression by PFF.

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Acknowledgments

The authors are grateful to Cor Semeins and Jolanda Hogervorst (Department of Oral Cell Biology, ACTA, Amsterdam, The Netherlands) and Carolina Sañudo (Department of Internal Medicine, H. U. Marqués de Valdecilla, IFIMAV, University of Cantabria, Spain) for their expert technical assistance This work was supported in part by grants from Instituto de Salud Carlos III–Fondo de Investigaciones Sanitarias (Spanish Ministry of Health) PI12/615 and SEIOMM-FEIOMM 2011. J.D.-C. was recipient of a fellowship from IFIMAV.

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  1. Jesús Delgado-Calle

    Present address: Department of Anatomy and Cell Biology, Indiana University School of Medicine, Indianapolis, IN, USA

Authors and Affiliations

  1. Department of Internal Medicine, Hospital U. M. Valdecilla-IFIMAV, University of Cantabria, RETICEF, Av., Marqués de Valdecilla, s/n, 39008, Santander, Spain

    Jesús Delgado-Calle & José A. Riancho

  2. Department of Oral Cell Biology, Academic Centre for Dentistry Amsterdam (ACTA), University of Amsterdam and VU University Amsterdam, Research Institute MOVE, Amsterdam, The Netherlands

    Jenneke Klein-Nulend

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  1. Jesús Delgado-Calle
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  2. José A. Riancho
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  3. Jenneke Klein-Nulend
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Correspondence to José A. Riancho.

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All authors have read and approve this version of the article, and due care has been taken to ensure the integrity of the work. No part of this paper has published or submitted elsewhere. No conflict of interest exits in the submission of this manuscript.

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Delgado-Calle, J., Riancho, J.A. & Klein-Nulend, J. Nitric Oxide is Involved in the Down-regulation of SOST Expression Induced by Mechanical Loading. Calcif Tissue Int 94, 414–422 (2014). https://doi.org/10.1007/s00223-013-9821-8

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