VDR dependent and independent effects of 1,25-dihydroxyvitamin D 3 on nitric oxide production by osteoblasts

Hubertine Willems, Ellen G.H.M. Van Den Heuvel, Geert Carmeliet, Anne Schaafsma, Jenneke Klein-Nulend, Astrid D. Bakker

Research output: Contribution to journalArticle

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Abstract

1,25-Dihydroxyvitamin D 3 (1,25(OH) 2D 3) strongly mediates bone mass. Mechanical stimulation also affects bone mass, partly via enhancing nitric oxide (NO) production by osteoblasts. We aimed to determine whether 1,25(OH) 2D 3 affects NO production by osteoblasts in the presence or absence of mechanical stimulation. We hypothesised that 1,25(OH) 2D 3 stimulates NO production via nuclear actions of the vitamin D receptor (VDR), which requires hours of incubation with 1,25(OH) 2D 3 to occur. MC3T3-E1 osteoblasts and long-bone osteoblasts of adult wildtype and VDR -/- mice were pre-incubated for 24 h with or without 1,25(OH) 2D 3 (10 -13-10 -9 M), followed by 30 min pulsating fluid flow (PFF; 0.7 ± 0.3 Pa, 5 Hz) or static culture with or without 1,25(OH) 2D 3. NO production and NO synthase (NOS) expression were quantified. 10 -11 M 1,25(OH) 2D 3 for 24 h, but not 30 min, stimulated NO production by MC3T3-E1 osteoblasts (eightfold). 1,25(OH) 2D 3 for 24 h increased inducible-NOS gene-expression (twofold), suggesting that 1,25(OH) 2D 3 stimulated NO production via activation of NOS gene transcription. PFF rapidly increased NO production by MC3T3-E1 osteoblasts, wildtype osteoblasts, and VDR -/- osteoblasts. This PFF effect was abolished after incubation with 1,25(OH) 2D 3 for 24 h, or during PFF only. Our results suggest that 1,25(OH) 2D 3 stimulates inducible-NOS expression and NO production by osteoblasts in the absence of mechanical stimulation, likely via genomic VDR action. In contrast, 1,25(OH) 2D 3 may affect mechanical loading-induced NO production independent of genomic VDR action, since 1,25(OH) 2D 3 diminished PFF-induced NO production in VDR -/- bone cells. In conclusion, 1,25(OH) 2D 3 and mechanical loading interact at the level of mechanotransduction, whereby 1,25(OH) 2D 3 seems to act independently of VDR genomic mechanism.

Original languageEnglish (US)
Pages (from-to)126-131
Number of pages6
JournalSteroids
Volume77
Issue number1-2
DOIs
StatePublished - Jan 1 2012

Fingerprint

Calcitriol Receptors
Osteoblasts
Nitric Oxide
Nitric Oxide Synthase
Bone
Bone and Bones
1,25-dihydroxyvitamin D
Pulsatile Flow
Transcription
Gene expression
Flow of fluids
Genes
Chemical activation
Gene Expression

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Endocrinology
  • Pharmacology
  • Clinical Biochemistry
  • Organic Chemistry

Cite this

Willems, H., Van Den Heuvel, E. G. H. M., Carmeliet, G., Schaafsma, A., Klein-Nulend, J., & Bakker, A. D. (2012). VDR dependent and independent effects of 1,25-dihydroxyvitamin D 3 on nitric oxide production by osteoblasts. Steroids, 77(1-2), 126-131. https://doi.org/10.1016/j.steroids.2011.10.015

VDR dependent and independent effects of 1,25-dihydroxyvitamin D 3 on nitric oxide production by osteoblasts. / Willems, Hubertine; Van Den Heuvel, Ellen G.H.M.; Carmeliet, Geert; Schaafsma, Anne; Klein-Nulend, Jenneke; Bakker, Astrid D.

In: Steroids, Vol. 77, No. 1-2, 01.01.2012, p. 126-131.

Research output: Contribution to journalArticle

Willems, H, Van Den Heuvel, EGHM, Carmeliet, G, Schaafsma, A, Klein-Nulend, J & Bakker, AD 2012, 'VDR dependent and independent effects of 1,25-dihydroxyvitamin D 3 on nitric oxide production by osteoblasts', Steroids, vol. 77, no. 1-2, pp. 126-131. https://doi.org/10.1016/j.steroids.2011.10.015
Willems, Hubertine ; Van Den Heuvel, Ellen G.H.M. ; Carmeliet, Geert ; Schaafsma, Anne ; Klein-Nulend, Jenneke ; Bakker, Astrid D. / VDR dependent and independent effects of 1,25-dihydroxyvitamin D 3 on nitric oxide production by osteoblasts. In: Steroids. 2012 ; Vol. 77, No. 1-2. pp. 126-131.
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AU - Klein-Nulend, Jenneke

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