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Effects of iguratimod on glucocorticoid-induced disorder of bone metabolism in vitro

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Journal of Bone and Mineral Metabolism Aims and scope Submit manuscript

Abstract

Introduction

Glucocorticoids are widely used to treat various diseases including rheumatoid arthritis (RA); however, one of the most frequent and severe adverse effects is glucocorticoid-induced osteoporosis (GIOP). Iguratimod (IGU) is a novel conventional synthetic disease-modifying anti-rheumatic drug developed in Japan. The aim of this study is to investigate the effects of IGU on glucocorticoid-induced disorder of bone metabolism in vitro.

Materials and methods

In osteoclastogenesis of mouse bone marrow-derived cells, tartrate-resistant acid phosphatase staining, resorption pit assay, western blotting, real-time polymerase chain reaction (PCR), and mRNA sequencing were performed. In osteoblastogenesis of MC3T3-E1 cells, alkaline phosphatase (ALP) staining and activity, alizarin red staining, and mRNA sequencing were performed, and real-time PCR and western blotting were conducted in MC3T3-E1 cells and murine osteocyte-like cell line MLO-Y4 cells.

Results

IGU significantly suppressed a dexamethasone-induced increase in osteoclasts, differentiation, and bone resorption activity by inhibition of the receptor activator of the nuclear factor kappa-B (RANK)/tumor necrosis factor receptor (TNFR)-associated factor 6 (TRAF6)/nuclear factor kappa-B (NFκB)-p52 pathway. In MC3T3-E1 cells, IGU significantly upregulated dexamethasone-induced downregulation of ALP activity, bone mineralization, and osteoblast-related gene and protein expression. In MLO-Y4 cells, IGU significantly upregulated dexamethasone-induced downregulation of the gene expression of ALP and osteocalcin, and also downregulated receptor activator of NFκB ligand (RANKL)/osteoprotegerin gene expression ratio without dexamethasone.

Conclusion

These results suggest that IGU may improve glucocorticoid-induced disorder of bone metabolism and may exhibit positive effects against GIOP associated with RA.

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Acknowledgements

We would like to thank F. Hirayama and Y. Eguchi for their excellent technical assistance.

Author information

Authors and Affiliations

  1. Department of Orthopaedic Surgery, Osaka University Graduate School of Medicine, 2-2 Yamada-oka, Suita, Osaka, 565-0871, Japan

    Akira Miyama, Makoto Hirao, Yuki Etani, Kenji Takami, Atsushi Goshima & Taihei Miura

  2. Department of Musculoskeletal Regenerative Medicine, Osaka University Graduate School of Medicine, 2-2 Yamada-oka, Suita, Osaka, 565-0871, Japan

    Kosuke Ebina & Shohei Oyama

  3. Department of Orthopaedic Surgery, Osaka Rosai Hospital, 1179-3 Nagasone-cho, Kita-ku, Sakai, 591-8025, Japan

    Gensuke Okamura

  4. Department of Health and Sport Sciences, Osaka University Graduate School of Medicine, 2-2 Yamada-oka, Suita, Osaka, 565-0871, Japan

    Takashi Kanamoto & Ken Nakata

  5. Department of Orthopaedic Surgery, Toyonaka Municipal Hospital, 4-14-1 Shibaharacho, Toyonaka, Osaka, 560-8565, Japan

    Hideki Yoshikawa

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  1. Akira Miyama
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Corresponding author

Correspondence to Kosuke Ebina.

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Conflict of interest

The iguratimod was kindly provided by Toyama Chemical Co., Ltd (Tokyo, Japan). K. Ebina has received research grants and lecture fee from Eisai Co., Ltd. K. Ebina and S. Oyama are affiliated with, and K. Nakata supervises the Department of Musculoskeletal Regenerative Medicine, Osaka University Graduate School of Medicine, which is supported by Taisho Pharmaceutical Co., Ltd. S. Oyama is an employee of Taisho Pharmaceutical Co., Ltd. These companies had no role in the study design, decision to publish, or preparation of the manuscript. A. Miyama, M. Hirao, G. Okamura, Y. Etani, K. Takami, A. Goshima, T. Miura, T. Kanamoto, and H. Yoshikawa declare that they have no conflicts of interest.

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774_2021_1206_MOESM1_ESM.doc

Supplementary file1 Supplementary Fig. KEGG map of parathyroid hormone activated pathway in comparison with or without IGU in the absence of Dex. Red rectangles represent significantly upregulated factors. Dex, dexamethasone; IGU, iguratimod; PTH, parathyroid hormone; KEGG, Kyoto Encyclopedia of Genes and Genomes. (DOC 41 KB)

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Miyama, A., Ebina, K., Hirao, M. et al. Effects of iguratimod on glucocorticoid-induced disorder of bone metabolism in vitro. J Bone Miner Metab 39, 639–648 (2021). https://doi.org/10.1007/s00774-021-01206-5

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