Idebenone attenuates RANKL-induced osteoclastogenesis and improves bone mass in ovariectomized mice

Postmenopausal osteoporosis (PMOP), driven by estrogen deficiency-induced osteoclast overactivation, poses a significant global health burden with limited therapeutic options. In this study, we investigated the potential of Idebenone (IDB) as a therapeutic agent for PMOP, focusing on its effects on osteoclast differentiation and underlying molecular mechanisms. Bone marrow-derived macrophages (BMMs) were cultured and stimulated to induce osteoclast differentiation, with or without IDB treatment. IDB's effects on osteoclast formation were assessed via TRAcP staining, F-actin ring visualization, resorption pit assays, and Western blotting. Network pharmacology highlighted the protein kinase B (AKT) as a key target, corroborated by molecular docking and CETSA showing IDB-AKT interaction. We found that IDB suppressed osteoclast differentiation and activity through reducing oxidative stress and inhibiting the AKT and mitogen-activated protein kinase (MAPK) signaling pathways. In vivo, ovariectomized (OVX) mice were treated with IDB (50 mg/kg) for six weeks, followed by micro-CT and histomorphometric analysis to evaluate bone microstructure and osteoclast activity. Notably, in OVX mice, IDB treatment significantly improved bone mass compared to controls. These findings suggested that IDB may serve as a promising therapeutic candidate for the treatment of postmenopausal osteoporosis by modulating osteoclast differentiation.