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Effects of the 1, 4-dihydropyridine L-type calcium channel blocker benidipine on bone marrow stromal cells

Abstract

Osteoporosis (OP) often increases the risk of bone fracture and other complications and is a major clinical problem. Previous studies have found that high blood pressure is associated with bone formation abnormalities, resulting in increased calcium loss. We have investigated the effect of the antihypertensive drug benidipine on bone marrow stromal cell (BMSC) differentiation into osteoblasts and bone formation under osteoporotic conditions. We used a combination of in vitro and in vivo approaches to test the hypothesis that benidipine promotes murine BMSC differentiation into osteoblasts. Alkaline phosphatase (ALP), osteocalcin (OCN), runt-related transcription factor 2 (RUNX2), β-catenin, and low-density lipoprotein receptor-related protein 5 (LRP5) protein expression was evaluated in primary femoral BMSCs from C57/BL6 mice cultured under osteogenic conditions for 2 weeks to examine the effects of benidipine. An ovariectomized (OVX) mouse model was used to investigate the effect of benidipine treatment for 3 months in vivo. We found that ALP, OCN, and RUNX2 expression was up-regulated and WNT/β-catenin signaling was enhanced in vitro and in vivo. In OVX mice that were intragastrically administered benidipine, bone parameters (trabecular thickness, bone mineral density, and trabecular number) in the distal femoral metaphysis were significantly increased compared with control OVX mice. Consistently, benidipine promoted BMSC differentiation into osteoblasts and protected against bone loss in OVX mice. Therefore, benidipine might be a suitable candidate for the treatment of patients with postmenopausal osteoporosis and hypertension.

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Acknowledgments

We thank Hua Wu (Hohhot Zhongke Medical Technology, Inner Mongolia, PR China) for excellent technical support with micro-CT.

Author information

Correspondence to Dao-zhang Cai.

Additional information

Z-p.M. and J-c.L. contributed equally to this work.

Design of the study: Z-p.M., J-c.L., and D-z.C. Acquisition of data: Z-p.M., J-c.L., D-z.C., and C.Z. Interpretation of data: Z-p.M., J-c.L., D-z.C., and C.Z. Manuscript preparation: Z-p.M., J-c.L., D-z.C., and C.Z. The authors declare no competing interests. This work was not commisioned and was externally peer-reviewed. Ethical approval was given by the Medical Ethics Committee of Inner Mongolia Medical University.

This project was funded by the Inner Mongolia Medical University Science and Technology Project (YKD2012KJBW003).

Electronic supplementary material

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Supplementary Fig. 1
figure7

BD is not able to promote changes in osteogenic markers such as OCN (b) and RUNX2 (c) in the absence of differentiation medium for 2 weeks. Columns represent the means ± SD from three wells per group (b, c). (GIF 29 kb)

Supplementary Fig. 2
figure8

BD has no effect on endogenous stem cells. As shown by the results of immunohistochemistry with the characteristic markers of marrow stromal cells, such as CD105 (a) and CD106 (b), endogenous stem cells do not change significantly after BD treatment. CD105- (c) and CD106- (d) positive cells are evenly distributed in all groups as demonstrated microscopically by immunohistochemical staining, with no significant differences between each other. Columns represent the means ± SD from six mice per group (c, d). (GIF 218 kb)

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Ma, Z., Liao, J., Zhao, C. et al. Effects of the 1, 4-dihydropyridine L-type calcium channel blocker benidipine on bone marrow stromal cells. Cell Tissue Res 361, 467–476 (2015). https://doi.org/10.1007/s00441-015-2115-x

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Keywords

  • Osteoblast
  • Benidipine
  • WNT signal pathway
  • Bone metabolism
  • Osteoporosis