Eupatilin Inhibits RANKL-induced Osteoclast Differentiation by Regulating MAPK and NF-κB Signaling Pathways
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 concentra-tions of Eupatilin on the cell activity of osteoclast precursor cells RAW264.7 and mouse bone marrow-derived mac-rophages(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 stai-ning 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 inhibito-ry effect on RANKL-induced osteoclast differentiation by regulating the MAPK and NF-κB signaling pathways.
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