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Inhibition of the classical NF-κB pathway prevents osteoclast bone-resorbing activity

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Abstract

The classical NF-κB pathway plays an important role in osteoclast formation and differentiation; however, the role of NF-κB in osteoclast bone-resorbing activity is not well understood. To elucidate whether NF-κB is important for osteoclast bone-resorbing activity, we used a selective peptide inhibitor of the classical NF-κB pathway named the NBD peptide. Osteoclasts were generated using bone marrow macrophages in the presence of M-CSF and RANKL. The NBD peptide dose-dependently blocked the bone-resorbing activity of osteoclasts by reducing area, volume (p < 0.001) and depths (p < 0.05) of pits. The reduced resorption by the peptide was due to reduced osteoclast bone-resorbing activity, but not reduced differentiation as the number of osteoclasts was similar in all groups. The peptide inhibited bone resorption by reducing TRAP activity, disrupting actin rings and preventing osteoclast migration. Gene expressions of a panel of bone resorption markers were significantly reduced. The NBD peptide dose-dependently reduced the RANKL-induced c-Src kinase activity, which is important for actin ring formation and osteoclast bone resorption. Therefore, these data suggest that the classical NF-κB pathway plays a pivotal role in osteoclast bone-resorbing activity.

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Acknowledgments

We would like to thank Dr. Michael J. May, University of Pennsylvania, for providing us with the NBD peptide and the immense help in preparation of this manuscript. We also would like to thank Dr. So Aoki for the preliminary experiment on this study. This study was supported by the grants from the Ministry of Education, Culture, Sports, Science and Technology of Japan, to H.S. (18390497), K.A. (19390472) and K.O. (19390471).

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Authors and Affiliations

  1. Section of Pharmacology, Department of Hard Tissue Engineering, Graduate School, Tokyo Medical and Dental University, 1-5-45 Yushima, Bunkyo-ku, Tokyo, 113-8549, Japan

    Niroshani S. Soysa, Neil Alles, Hitoyata Shimokawa, Kazuhiro Aoki & Keiichi Ohya

  2. Center of Excellence Program for Frontier Research on Molecular Destruction and Reconstruction of Tooth and Bone, Tokyo Medical and Dental University, Tokyo, Japan

    Niroshani S. Soysa & Neil Alles

  3. Division of Molecular Biochemistry, Department of Biosciences, Kyushu Dental College, Kitakyushu, Japan

    Eijiro Jimi

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  1. Niroshani S. Soysa
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Correspondence to Kazuhiro Aoki.

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Soysa, N.S., Alles, N., Shimokawa, H. et al. Inhibition of the classical NF-κB pathway prevents osteoclast bone-resorbing activity. J Bone Miner Metab 27, 131–139 (2009). https://doi.org/10.1007/s00774-008-0026-6

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  • DOI: https://doi.org/10.1007/s00774-008-0026-6

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