Carnosol inhibits osteoclastogenesis in vivo and in vitro by blocking the RANKL‑induced NF‑κB signaling pathway

Mol Med Rep. 2022 Jul;26(1):225. doi: 10.3892/mmr.2022.12741. Epub 2022 May 20.

Abstract

Bone homeostasis is maintained by osteoclast-mediated bone resorption and osteoblast‑mediated bone formation. Disruption of bone homeostasis due to excessive osteoclastogenesis or reduced osteogenesis results in various disorders, such as postmenopausal osteoporosis. Receptor activator of NF‑κB ligand (RANKL) stimulation of the NF‑κB signaling pathway is essential in osteoclastogenesis. The aim of the present study was to investigate the novel effects of carnosol, an active compound found in Rosmarinus officinalis, on RANKL‑induced osteoclastogenesis both in vitro and in vivo. TRAP staining showed that carnosol significantly inhibited osteoclasts differentiation of bone marrow monocytes and RAW264.7 cells. Western blot results showed that the protein expression levels of osteoclastogenesis‑associated genes, including cathepsin K, tartrate‑resistant acid phosphatase and MMP‑9, were markedly inhibited by carnosol, which may have suppressed osteoclast function. Furthermore, western blot and immunofluorescent staining results revealed that carnosol markedly suppressed the phosphorylation of p65 induced by RANKL and blocked its nuclear translocation, indicating the suppression of NF‑κB signaling pathway. H&E staining and micro‑CT results showed that in vivo treatment with carnosol significantly attenuated ovariectomy‑induced bone loss in mice. In conclusion, the present study indicated that carnosol may suppress osteoclastogenesis both in vivo and in vitro by inhibiting the activation of the NF‑κB signaling pathway. Carnosol may therefore be a potential novel therapeutic candidate for the clinical treatment of osteoclast‑related disorders.

Keywords: NF‑κB; PMOP; RANKL; carnosol; osteoclastogenesis.

MeSH terms

  • Abietanes
  • Animals
  • Bone Resorption* / metabolism
  • Cell Differentiation
  • Female
  • Humans
  • Mice
  • NF-kappa B / metabolism
  • Osteoclasts / metabolism
  • Osteogenesis*
  • RANK Ligand / metabolism
  • RANK Ligand / pharmacology
  • RAW 264.7 Cells
  • Signal Transduction

Substances

  • Abietanes
  • NF-kappa B
  • RANK Ligand
  • carnosol

Grants and funding

This present study was supported by the Construction of Key Medical Specialties in Shanghai (grant no. ZK2019B05), the Shanghai Pudong New Area Health System Key Discipline Group (grant no. PWZxq2017-12), Clinical Characteristics of Health System in Pudong New Area (grant no. PWYts2021-03), Pudong New Area of Shanghai Characteristic Treatment of Degenerative Lumbar Instability in Old Age (grant no. PWZzb2017-33), the Medical Science and Technology Development Project (grant no. 2017-YKK17250) and the Promotion Project of Advanced and Appropriate Technology from Shanghai Healthcare Commission (grant no. 2019SY069).