Belamcandin inhibits osteoclast differentiation: its role and mechanism

doi:10.12307/2022.527

Abstract

Abstract: BACKGROUND: Receptor activator of nuclear factor kappa B ligand can mediate the activation of calcium/nuclear factor of activated T-cells cytoplasmic 1 (NFATc1) pathway, which is crucial for osteoclast differentiation. Belamcandin can exert estrogen-like effects by activating estrogen receptors; however, less is reported on its effects on the differentiation and function of osteoclasts.
OBJECTIVE: To explore the effect of Belamcandin on osteoclasts and its mechanism.
METHODS: Mature mouse bone marrow macrophages cultured in vitro were differentiated and seeded into 96-well plates (6×103 cells per well), followed by inhibitory interventions with 5, 10, 20, 30, 40 μmol/L Belamcandin. Osteocyte differentiation was then observed to screen out the optimal concentration of Belamcandin with the strongest effect. Different concentrations of Belamcandin were used to deal with the differentiation of osteoclasts induced by nuclear factor κB receptor activator ligands. Tartrate-resistant acid phosphatase staining was used to observe the effects of Belamcandin on the formation and function of osteoclasts. Expression of upstream and downstream genes and proteins in the Calcium/NFATc1 signaling pathway was observed through calcium ion oscillation experiments, western blot assay and RT-qPCR detection. An approval was obtained from the Animal Experiment Ethics Committee of the First Clinical Medical College of Guangzhou University of Chinese Medicine.
RESULTS AND CONCLUSION: Belamcandin inhibited the formation of osteoclasts in a dose-dependent manner, and 40 μmol/L Belamcandin had the strongest potential to inhibit osteoclast differentiation. Belamcandin could inhibit the mRNA expression of Acp5 (tartrate resistant acid phosphatase) gene related to the inhibition of osteoclast formation and the protein expression of c-Fos and Integrin β3, while inhibiting the expression of Ctsk, matrix metalloproteinase 9 gene, and transcription protein related to the inhibition of bone resorption function. Belamcandin could inhibit the opening frequency and intensity of calcium ion signal channels, and restrain the expression of Calcitonin receptor, Nfatc1 gene and transcription protein. To conclude, Belamcandin can inhibit the expression of Nfatc1 gene and transcription protein through the calcium ion signaling pathway induced by nuclear factor κB receptor activator ligand, thereby inhibiting osteoclast formation and bone resorption.

Key words: Belamcandin, osteoclasts, calcium ions, NFATc1, cell differentiation

CLC Number:

Huang Fayi, Liu Yuhao, Zhou Chi, Huang Haoran, Chen Weijian, He Wei, Wang Haibin. Belamcandin inhibits osteoclast differentiation: its role and mechanism[J]. Chinese Journal of Tissue Engineering Research, 2022, 26(17): 2636-2641.

Figures/Tables 5

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