Eupatilin Inhibits RANKL-induced Osteoclast Differentiation by Regulating MAPK and NF-κB Signaling Pathways

doi:10.13701/j.cnki.kqyxyj.2024.09.012

Abstract

Abstract: Objective: To investigate the effects and molecular mechanisms of eupatilin on the differentiation of osteoclast precursor cells to osteoblasts. Methods: CCK-8 assay was used to detect the effects of different concentrations of Eupatilin on the cell activity of osteoclast precursor cells RAW264.7 and mouse bone marrow-derived macrophages (BMDMs). RAW264.7 and BMDMs were induced to differentiate into osteoclasts by receptor activator of nuclear factor-κB ligand (RANKL), and different concentrations of Eupatilin (5, 10, and 20 μmol/L) were used for intervention. The effects of Eupatilin on RANKL-induced osteoclast formation were evaluated through TRAP staining and F-actin staining. The expression of osteoclast-related marker genes was detected by qRT-PCR and Western blot, and the phosphorylation levels of mitogen-activated protein kinase (MAPK) and nuclear factor-κB (NF-κB) signaling pathway molecules were detected by Western blot. Results: Eupatilin at 20 μmol/L exhibited a significant inhibitory effect on the differentiation of osteoclasts, and also shown an effective downregulation of the expression of related marker genes such as tartrate-resistant acid phosphas (TRAP), matrix metalloproteinase-9 (MMP-9), and cathepsin K (CTSK). Western blot results indicated that Eupatilin significantly inhibited the phosphorylation levels of MAPK and NF-κB signaling pathway molecules induced by RANKL. Conclusion: Eupatilin exerts a significant inhibitory effect on RANKL-induced osteoclast differentiation by regulating the MAPK and NF-κB signaling pathways.

Key words: Eupatilin, osteoclast, MAPK signaling pathway, NF-κB signaling pathway

ZHAO Boxuan, LI Ting, YAO Hantao, WANG Zijun, GUO Haiying, JI Yaoting, DU Minquan. Eupatilin Inhibits RANKL-induced Osteoclast Differentiation by Regulating MAPK and NF-κB Signaling Pathways[J]. Journal of Oral Science Research, 2024, 40(9): 820-826.

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