Abstract
BackgroundSophorae Flos (SF) is a composite of flowers and buds of Styphnolobium japonicum (L.) Schott and has been used in traditional Korean and Chinese medicine for the treatment of hemostasis and inflammation. Previous studies reported that SF possesses anti-obesity properties, as well as anti-allergic, anti-proliferative, and anti-inflammatory activities. However, the effect of SF in bone resorption has not been studies. In this study, we examined the potential of SF extract (SFE) to inhibit receptor activator of NF-κB ligand (RANKL) -induced osteoclast differentiation in cultured mouse-derived bone marrow macrophages (BMMs).MethodsBMMs, that act as osteoclast precursors, were cultured with M-CSF (50 ng/ml) and RANKL (100 ng/ml) for 4 days to generate osteoclasts. Osteoclast differentiation was measured by tartrate-resistant acidic phosphatase (TRAP) staining and the TRAP solution assay. Osteoclast differentiation marker genes were analyzed by the quantitative real-time polymerase chain reaction analysis. RANKLs signaling pathways were confirmed through western blotting.ResultsSFE significantly decreased osteoclast differentiation in a dose-dependent manner. SFE inhibited RANKL-induced osteoclastogenesis by suppressing NF-κB activation. By contrast, SFE did not affect phospholipase C gamma 2 or subsequent cAMP response element binding activation. SFE inhibited the RANKL-induced expression of nuclear factor of activated T cells c1 (NFATc1).ConclusionsSFE attenuated the RANKL-mediated induction of NF-κB through inhibition of IκBα phosphorylation, which contributed to inhibiting of RANKL-induced osteoclast differentiation through downregulation of NFATc1.
Highlights
Sophorae Flos (SF) is a composite of flowers and buds of Styphnolobium japonicum (L.) Schott and has been used in traditional Korean and Chinese medicine for the treatment of hemostasis and inflammation
Thereafter, the RANK ligand (RANKL)/Receptor activator of NF-κB (RANK) interaction activates extracellular signal-regulated kinase (ERK), p38, c-Jun N-terminal kinase (JNK), Akt, and NF-κB [7,8,9]. These signal transduction pathways lead to the expression and activation of transcription factors such as nuclear factor of activated T cells c1 (NFATc1) and activator protein-1 (AP-1), both of which are involved in the expression of genes specific to osteoclasts [9,10,11]
Cytotoxic effects of SF extract (SFE) To investigate the cytotoxicity of SFE on bone marrow macrophages (BMMs), it treated with several concentrations of SFE (0, 25, 50, 100, and 200 μg/ml) for 1 day
Summary
Sophorae Flos (SF) is a composite of flowers and buds of Styphnolobium japonicum (L.) Schott and has been used in traditional Korean and Chinese medicine for the treatment of hemostasis and inflammation. Osteoclasts are specialized as the only bone-resorbing cell type and increased numbers are implicated in the development of bone loss-accompanied diseases such as Macrophage colony-stimulating factor (M-CSF) and receptor activator of NF-κB ligand (RANKL) are two cytokines secreted mainly by osteoclast Both are differentiation factors [5, 6]. Thereafter, the RANKL/RANK interaction activates extracellular signal-regulated kinase (ERK), p38, c-Jun N-terminal kinase (JNK), Akt, and NF-κB [7,8,9] These signal transduction pathways lead to the expression and activation of transcription factors such as nuclear factor of activated T cells c1 (NFATc1) and activator protein-1 (AP-1), both of which are involved in the expression of genes specific to osteoclasts [9,10,11]
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