Melatonin inhibits RANKL-induced osteoclastogenesis through the miR-882/Rev-erbα axis in Raw264.7 cells

Melatonin, secreted in a typical diurnal rhythm pattern, has been reported to prevent osteoporosis; however, its role in osteoclastogenesis remains unclear. In the present study, the ability of melatonin to inhibit receptor activator of nuclear factor-κB ligand (RANKL)-induced osteoclastogenesis and the associated mechanism were investigated. Raw264.7 cells were cultured with RANKL (100 ng/ml) and macrophage colony-stimulating factor (M-CSF; 30 ng/ml) for 7 days, and tartrate-resistant acid phosphatase (TRAP) staining was used to detect osteoclastogenesis following treatment with melatonin. In addition, the effect of melatonin on cathepsin K and microRNA (miR)-882 expression was investigated via western blotting and reverse transcription-quantitative PCR. Melatonin significantly inhibited RANKL-induced osteoclastogenesis in Raw264.7 cells. From bioinformatics analysis, it was inferred that nuclear receptor subfamily 1 group D member 1 (NR1D1/Rev-erbα) may be a target of miR-882. In vitro, melatonin upregulated Rev-erbα expression and downregulated miR-882 expression in the osteoclastogenesis model. Rev-erbα overexpression boosted the anti-osteoclastogenesis effects of melatonin, whereas miR-882 partially diminished these effects. The present results indicated that the miR-882/Rev-erbα axis may serve a vital role in inhibiting osteoclastogenesis following RANKL and M-CSF treatment, indicating that Rev-erbα agonism or miR-882 inhibition may represent mechanisms through which melatonin prevents osteoporosis.