Abstract

BackgroundDestructive erosion of bone or osteolysis is a major complication of inflammatory conditions such as rheumatoid arthritis (RA), periodontal disease, and periprosthetic osteolysis. Natural plant-derived products have received recent attention as potential therapeutic and preventative drugs in human disease.MethodsThe effect of Angelica sinensis (AS) extract on RANKL-induced osteoclast differentiation was examined in this study. The osteoclast precursor cell line bone marrow macrophages (BMMs) was cultured and stimulated with RANKL followed by treatment with AS at several doses. Gene expression profiles of c-Fos, c-Jun, NFATc1, TRAP, and OSCAR were sequentially evaluated.ResultsAS extract inhibited RANKL-mediated osteoclast differentiation in BMMs in a dose-dependent manner without any evidence of cytotoxicity. AS extract strongly inhibited p38, ERK, JNK, p65 phosphorylation and I-κB degradation in RANKL-stimulated BMMs. AS extract also inhibited the mRNA expression of c-Fos, c-Jun, NFATc1, TRAP, and OSCAR in RANKL-treated BMMs. Moreover, RANKL-induced c-Fos, c-Jun and NFATc1 protein expression was suppressed by AS extract.ConclusionsThese results collectively suggested that AS extract demonstrated inhibitory effects on RANKL-mediated osteoclast differentiation in bone marrow macrophages in vitro, indicating that AS may therefore serve as a useful drug in the prevention of bone loss.

Highlights

  • Destructive erosion of bone or osteolysis is a major complication of inflammatory conditions such as rheumatoid arthritis (RA), periodontal disease, and periprosthetic osteolysis

  • Antibodies for cFos, c-Jun and nuclear factor of activated T cells 1 (NFATc1) were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA), and Western blot antibodies for phosphor-p65, p-65, phosphor-ERK, ERK, phosphorJNK, JNK, phosphor-p38, p38, and I-κB were from Santa Cruz Biotechnology Inc. (Santa Cruz, CA, USA); β-actin antibody was purchased from Sigma-Aldrich, Inc

  • Inhibition of osteoclast differentiation by Angelica sinensis (AS) extract Osteoclasts were generated from mouse bone marrow macrophages (BMMs) in the presence of M-CSF (20 ng/ml) plus RANKL (40 ng/ml) to verify the effects of AS extract in osteoclastogenesis

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Summary

Introduction

Destructive erosion of bone or osteolysis is a major complication of inflammatory conditions such as rheumatoid arthritis (RA), periodontal disease, and periprosthetic osteolysis. Natural plant-derived products have received recent attention as potential therapeutic and preventative drugs in human disease. M-CSF induces expression of RANKL receptor (RANK) as well as supports survival and proliferation in early stage precursors of osteoclast lineages in mouse bone marrow cells [3]. RANKL is a member of the tumor necrosis factor (TNF) family and binds to the RANK receptor expressed in osteoclast precursor cells [4]. Several compounds derived from natural products have been recently reported to possess inhibitory effects on osteoclast differentiation and function, leading to decreased bone loss in vivo [1]. We aimed to investigate the effects of AS extract on signaling pathways involved in osteoclast differentiation, activation, and survival in vitro

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