Obacunone targets macrophage migration inhibitory factor (MIF) to impede osteoclastogenesis and alleviate ovariectomy-induced bone loss

• Published in: Journal of advanced research 2023-11, Vol.53, p.235-248
• Main Authors: He, Jianbo, Zheng, Lin, Li, Xiaojuan, Huang, Furong, Hu, Sitao, Chen, Lei, Jiang, Manya, Lin, Xianfeng, Jiang, Haibo, Zeng, Yifan, Ye, Tianshen, Lin, Dingkun, Liu, Qian, Xu, Jiake, Chen, Kai
• Format: Article
• Language: English
• Subjects: Bone loss; Macrophage migration inhibitory factor (MIF); Obacunone; Osteoclast; Reactive oxygen species (ROS)
• ISSN: 2090-1232; 2090-1224
• DOI: 10.1016/j.jare.2023.01.003

Obacunone (OB) targets macrophage migration inhibitory factor (MIF) to impede osteoclastogenesis and alleviate osteoporosis. [Display omitted] •Obacunone (OB) effectively inhibits osteoclast formation and function in vitro.•OB attenuates RANKL-induced osteoclast differentiation signaling pathways.•Macrophage migration inhibitory factor (MIF) is identified as the molecular target of OB.•OB alleviates estrogen deficiency-induced bone loss by impeding excessive osteoclast activity.•OB may serve as an alternative therapeutic candidate for osteoporosis. Osteoporosis is the most common bone disorder where the hyperactive osteoclasts represent the leading role during the pathogenesis. Targeting hyperactive osteoclasts is currently the primary therapeutic strategy. However, concerns about the long-term efficacy and side effects of current frontline treatments persist. Alternative therapeutic agents are still needed. Obacunone (OB) is a small molecule with a broad spectrum of biological activities, particularly antioxidant and anti-inflammatory effects. This study aims to examine OB’s therapeutic potential on osteoporosis and explore the rudimentary mechanisms. Osteoclast formation and osteoclastic resorption assays were carried out to examine OB’s inhibitory effects in vitro, followed by the in-vivo studies of OB’s therapeutic effects on ovariectomy-induced osteoporotic preclinical model. To further study the underlying mechanisms, mRNA sequencing and analysis were used to investigate the changes of downstream pathways. The molecular targets of OB were predicted, and in-silico docking analysis was performed. Ligand-target binding was verified by surface plasmon resonance (SPR) assay and Western Blotting assay. The results indicated that OB suppressed the formation of osteoclast and its resorptive function in vitro. Mechanistically, OB interacts with macrophage migration inhibitory factor (MIF) which attenuates receptor activator of nuclear factor kappa B (NF-κB) ligand (RANKL)-induced signaling pathways, including reactive oxygen species (ROS), NF-κB pathway, and mitogen-activated protein kinases (MAPKs). These effects eventually caused the diminished expression level of the master transcriptional factor of osteoclastogenesis, nuclear factor of activated T cells 1 (NFATc1), and its downstream osteoclast-specific proteins. Furthermore, our data revealed that OB alleviated estrogen deficiency-induced osteoporosis by targeting MIF and thus inhibiting hyperactive oste