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Quercetin prevents bone loss in hindlimb suspension mice via stanniocalcin 1-mediated inhibition of osteoclastogenesis

Abstract

Recent studies demonstrate that diet quercetin (Quer) has obvious bone protective effects on ovariectomized rodents but thus far there is no direct evidence to support the inhibitory effect of Quer on bone loss caused by long-term unloading. In the present study, we investigated whether Quer could prevent bone loss induced by unloading in mice. Mice were subjected to hindlimb suspension (HLS) and received Quer (25, 50, 100 mg· kg−1 ·day−1, ig) for 4 weeks. Before euthanasia blood sample was collected; the femurs were harvested and subjected to MicroCT analysis. We showed that Quer administration markedly improved bone microstructure evidenced by dose-dependently reversing the reduction in bone volume per tissue volume, trabecular number, and bone mineral density, and the increase of trabecular spacing in mice with HLS. Analysis of serum markers and bone histometric parameters confirmed that Quer at both middle and high doses significantly decreased bone resorption-related markers collagen type I and tartrate-resistant acid phosphatase 5b, and increased bone formation-related marker procollagen 1 N-terminal propeptide as compared with HLS group. Treatment with Quer (1, 2, 5 μM) dose-dependently inhibited RANKL-induced osteoclastogenesis through promoting the expression of antioxidant hormone stanniocalcin 1 (STC1) and decreasing ROS generation; knockdown of STC1 blocked the inhibitory effect of Quer on ROS generation. Knockdown of STC1 also significantly promoted osteoclastogenesis in primary osteoclasts. In conclusion, Quer protects bones and prevents unloading-caused bone loss in mice through STC1-mediated inhibition of osteoclastogenesis. The findings suggest that Quer has the potential to prevent and treat off-load bone loss as an alternative supplement.

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Fig. 1: Quer, 3,5,7,3’,4’-pentahydroxy flavone used in this study.
Fig. 2: Quer treatment suppressed bone loss induced by unloading.
Fig. 3: Effect of Quer treatment on serum chemical markers of bone resorption and bone formation in male mice subjected to hindlimb suspension (HLS) in a 4-week trial that were sacrificed at 13 weeks of age.
Fig. 4: Effect of Quer treatment on femoral biomechanics as assessed by three-point bending in male mice subjected to hindlimb suspension (HLS) in a 4-week trial that were sacrificed at 13 weeks of age.
Fig. 5: Effect of Quer on bone formation and bone resorption as evaluated by bone histomorphometry parameters in male mice subjected to hindlimb suspension (HLS) for 4 weeks that were sacrificed at 13 weeks of age.
Fig. 6: Quer suppressed RANKL-induced osteoclastogenesis.
Fig. 7: Quer treatment inhibited RANKL-induced osteoclastogenesis via the STC1-mediated antioxidant pathway.
Fig. 8: Effects of Quer on STC1, NFATc1, and Ctsk expression and the osteoclastogenesis of osteoclasts induced to differentiate from primary bone marrow cells.

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Acknowledgements

This study was financially supported by grants from the Natural Science Foundation of Shaanxi Province (Nos. 2020JM-128 and 2019JM-260), the Fundamental Research Funds for the Central Universities (No. 3102017zy051), and the Graduate Creative Innovation Seed Fund of Northwestern Polytechnical University (No. ZZ2019271).

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YBN, YHL, and QBM designed the experiments. YBN wrote the paper. YHL, and YMZ, revised the paper. YYY, XX, YS, YHZ, and DD, performed the experiments. YYY, XX, CRL, and XLW, analyzed the data.

Corresponding authors

Correspondence to Yu-hua Li or Qi-bing Mei.

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The authors declare no competing interests.

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Niu, Yb., Yang, Yy., Xiao, X. et al. Quercetin prevents bone loss in hindlimb suspension mice via stanniocalcin 1-mediated inhibition of osteoclastogenesis. Acta Pharmacol Sin 41, 1476–1486 (2020). https://doi.org/10.1038/s41401-020-00509-z

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Keywords

  • quercetin
  • bone loss
  • osteoclast
  • STC1
  • ROS
  • hindlimb suspension mice

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