Picroside II Inhibits RANKL‐Mediated Osteoclastogenesis by Attenuating the NF‐κB and MAPKs Signaling Pathway In Vitro and Prevents Bone Loss in Lipopolysaccharide Treatment Mice

Abstract

Picroside II, one of the major components isolated from the seed of natural plant picrorhiza, is widely used in traditional Chinese medicine. The present study was performed to define effects of picroside II on nuclear factor-kappaB ligand (RANKL)-stimulated osteoclast differentiation in vitro and on lipopolysaccharide (LPS)-induced bone loss in vivo. The bone marrow cells (BMMs) were harvested and induced with RANKL followed by treatment with picroside II at several doses, and the differentiation of osteoclasts from these cells was evaluated by tartrate-resistant acid phosphatase (TRAP) staining and resorption pit formation assay. The effects of picroside II on osteoclastogenesis were studied by examining RANKL-induced osteoclast F-actin ring formation and osteoclast bone resorption. Moreover, we explored the mechanisms of these downregulation effects by performed Western blotting and quantitative RT-PCR examination. Results demonstrated picroside II strongly inhibited RANKL-induced osteoclast formation when added during the early stage of BMMs cultures, suggesting that it acts on osteoclast precursors to inhibit RANKL/RANK signaling. Moreover, picroside II markedly decreased the phosphorylation of p38, ERK, JNK, p65, and I-κB degradation, and significantly suppressed c-Fos and nuclear factor of activated T-cells cytoplasmic 1 (NFATc1), both the key transcription factors during osteoclastogenesis. Furthermore, in vivo studies verified the bone protection effects of picroside II. These results collectively suggested that picroside II acted as an anti-resorption agent by blocking osteoclast activation. J. Cell. Biochem. 118: 4479-4486, 2017. © 2017 Wiley Periodicals, Inc.

Picrasidine I from Picrasma Quassioides Suppresses Osteoclastogenesis via Inhibition of RANKL Induced Signaling Pathways and Attenuation of ROS Production

“…Cytotoxicity of PI on BMMs were performed as our previously described [17, 18]. Briefly, BMMs were seeded at a density of 3.5×10 3 cells per well in 96-well plates with various concentrations of PI and incubated for 3 d in the presence of RANKL.…”

“…Mature OCs function evaluated by bone absorption assay [19] and F-actin ring immunofluorescence as previously described respectively [17]. Briefly, cells were treated with 50 ng/ml RANKL on 6-well plates pre-coated with collagen for 4 days.…”

Inhibition effects of a natural inhibitor on RANKL downstream cellular signalling cascades cross‐talking

“…Despite the lack of effort by most of the large pharmaceutical companies, natural product research has still been active during the past decade . Institutions mainly from China, Korea, India, Japan and United States keep engaging in the work of natural products study, for re‐establish natural products as a major source for potential drug discovery . Given that only limited biological properties or functional activities have been worked out for each natural product, there still has a “long way to go” for fully identifying the biological activities of natural products .…”

Chemical components, pharmacological actions, and clinical applications of Rhizoma Picrorhizae

“…Others reported that (a) picroside II had a certain bone protection effect and could be used as an osteoclast blocker against bone resorption (Yang, Gao, & Wang, ); (b) oral administration of RP had the antagonistic effect on gastric ulcer induced by hydrochloric acid‐ethanol in rats (Anandan, Rekha, & Saravanan, ); (c) the water extract of RP had the inhibitory effect on some superficial fungi at 5% concentration (Wang, Yang, & Yang, ); and (d) RP could also reduce the damage of kidney caused by metal cadmium poisoning (Wang et al, 2017). In addition, RP can also improve and prolong the ovarian function, treat and alleviate menopausal symptoms, improve depression, and so on (Zhang et al, ).…”