Aconine inhibits RANKL-induced osteoclast differentiation in RAW264.7 cells by suppressing NF-κB and NFATc1 activation and DC-STAMP expression

Abstract

Aim: Aconiti Lateralis Radix Preparata is a traditional Chinese medicine used to treat chronic arthritis and is highly effective against rheumatoid arthritis. However, the effects of aconine, a derivative of aconitum alkaloids, on osteoclasts, which can absorb bone, remain unknown. Here, we investigated the effects of aconine on osteoclast differentiation and bone resorption in vitro. Methods: The viability of mouse leukemic monocyte/macrophage cell line RaW264.7 was measured using CCK-8 assays. osteoclast differentiation was induced by incubation of RaW264.7 cells in the presence of RanKL, and assessed with TRaP staining assay. Bone resorption was examined with bone resorption pits assay. The expression of relevant genes and proteins was analyzed using RT-PCR and Western blots. The activation of NF-κB and nuclear factor of activated T-cells (NFAT) was examined using stable NF-κB and NFATc1 luciferase reporter gene systems, RT-PCR and Western blot analysis. Results: Aconine (0.125, 0.25 μmol/L) did not affect the viability of RAW264.7 cells, but dose-dependently inhibited RANKL-induced osteoclast formation and bone resorptive activity. Furthermore, aconine dose-dependently inhibited the RanKL-induced activation of NF-κB and NFATc1 in RAW264.7 cells, and subsequently reduced the expression of osteoclast-specific genes (c-Src, β3-Integrin, cathepsin K and MMP-9) and the expression of dendritic cell-specific transmembrane protein (DC-STAMP), which played an important role in cell-cell fusion. Conclusion: These findings suggest that aconine inhibits RANKL-induced osteoclast differentiation in RAW264.7 cells by suppressing the activation of NF-κB and NFATc1 and the expression of the cell-cell fusion molecule DC-STAMP.

Effects of omega‐3 fatty acids and aspirin on Porphyromonas gingivalis‐induced periodontitis in rats

“…Osteoclasts are differentiated from hematopoietic myeloid precursor cells of monocyte/macrophage lineage . The murine macrophage cell line RAW264.7 cells are commonly used as osteoclast precursors; these cells have become a valuable tool to study osteoclast differentiation . RANKL induces osteoclast differentiation, function, and survival of cells .…”

“…24 The murine macrophage cell line RAW264.7 cells are commonly used as osteoclast precursors; these cells have become a valuable tool to study osteoclast differentiation. 25,26 RANKL induces osteoclast differentiation, function, and survival of cells. 27 In our study, we found that omega-3 plus aspirin markedly attenuated the number of RANKL-stimulated TRAP (+) multinucleated cells in RAW264.7 cells.…”

Osthole‐loaded N‐octyl‐O‐sulfonyl chitosan micelles (NSC‐OST) inhibits RANKL‐induced osteoclastogenesis and prevents ovariectomy‐induced bone loss in rats

“…Specifically, 10 μL of CCK‐8 solution was added to each well of cells. After further incubation of the plate in an incubator at 37°C for 4 hours, the absorbance at 450 nm was measured using a microplate reader (Perkin Elmar) and then the cell viability was analysed according to formula as described …”

Herbal compounds for rheumatoid arthritis: Literatures review and cheminformatics prediction

“…RA belongs to the category of Bi Zheng in traditional Chinese medicine theory. A variety of traditional Chinese medicines resistant to Bi Zheng show the potentials to treat RA (Gao, Feng, Zhang, Wang, & Cui, ; Zeng et al, ). In addition, in recent years, traditional Chinese medicines with anti‐inflammatory, immunomodulatory, or cartilage‐protective effects have also been widely used in the treatment of RA (Pu et al, ; Zeng et al, ).…”

“…A variety of traditional Chinese medicines resistant to Bi Zheng show the potentials to treat RA (Gao, Feng, Zhang, Wang, & Cui, ; Zeng et al, ). In addition, in recent years, traditional Chinese medicines with anti‐inflammatory, immunomodulatory, or cartilage‐protective effects have also been widely used in the treatment of RA (Pu et al, ; Zeng et al, ). Therefore, the herbal compounds with anti‐RA activities were reviewed in this paper, and the cheminformatics tools were used to predict their drug‐likeness properties and pharmacokinetic parameters, so as to provide the references for their follow‐up researches and developments.…”