Molecular mechanisms of the inhibitory effects of jiangu granule-containing serum on RANKL-induced osteoclastogenesis

Postmenopausal osteoporosis (PMOP) is characterized by increased bone loss due to enhanced osteoclastogenesis and bone resorption. A Chinese herbal formula, jiangugranule (JG), exhibited great efficacy in the clinical treatment of PMOP. However, the molecular mechanisms underlying the therapeutic effects remain unclear. The present study aimed to examine the effects of JG‑containing serum on receptor activator of nuclear factor‑κB (NF‑κB) ligand (RANKL)‑induced osteoclastogenesis. Osteoclast precursor RAW264.7 cells were cultured and treated with JG‑containing serum in the presence of RANKL. Following 6 days of culture, the cells were stained with tartrate‑resistant acid phosphatase and the rate of differentiation was calculated. In addition, cells were treated with JG‑containing serum for 24, 48 and 96 h and total RNA and proteins were extracted for reverse transcription‑quantitative polymerase chain reaction and western blot analysis to detect mRNA and protein expression, respectively, of key molecules in the RANK/RANKL signaling pathway, including RANK, tumor necrosis factor receptor‑associated factor 6, NF‑κB (p50 and p52 subunits), c‑Fos and nuclear factor of activated T cells, cytoplasmic 1 (NFATc1). The results revealed that JG‑containing serum inhibited RANKL‑induced osteoclastogenesis and reduced mRNA and protein expression of RANK, c‑Fos and NFATc1. The results suggested that JG may regulate osteoclast differentiation through the RANK/RANKL signaling pathway, which may be a possible mechanism for the therapeutic effects of JG on PMOP.