Cholesterol sulfate inhibits osteoclast differentiation and survival by regulating the AMPK–Sirt1–NF‐κB pathway

Abstract Cholesterol sulfate (CS) is an activator of retinoic acid‐related orphan receptor α (RORα). CS treatment or RORα overexpression attenuates osteoclastogenesis in a collagen‐induced arthritis mouse model. However, the mechanism by which CS and RORα regulate osteoclast differentiation remains largely unknown. Thus, we aimed to investigate the role of CS and RORα in osteoclastogenesis and their underlying mechanism. CS inhibited osteoclast differentiation, but RORα deficiency did not affect osteoclast differentiation and CS‐mediated inhibition of osteoclastogenesis. CS enhanced adenosine monophosphate‐activated protein kinase (AMPK) phosphorylation and sirtuin1 (Sirt1) activity, leading to nuclear factor‐κB (NF‐κB) inhibition by decreasing acetylation at Lys310 of p65. The NF‐κB inhibition was restored by AMPK inhibitor, but the effects of CS on AMPK and NF‐κB were not altered by RORα deficiency. CS also induced osteoclast apoptosis, which may be due to sustained AMPK activation and consequent NF‐κB inhibition, and the effects of CS were significantly reversed by interleukin‐1β treatment. Collectively, these results indicate that CS inhibits osteoclast differentiation and survival by suppressing NF‐κB via the AMPK–Sirt1 axis in a RORα‐independent manner. Furthermore, CS protects against bone destruction in lipopolysaccharide‐ and ovariectomy‐mediated bone loss mouse models, suggesting that CS is a useful therapeutic candidate for treating inflammation‐induced bone diseases and postmenopausal osteoporosis.