Function studies of pectolinarigenin demonstrated that, as a natural product, it possesses the regulatory effects on transcription factors (TFs) such as: signal transducer and activator of transcription 3 (STAT3). Herein, we aimed to identify the regulatroy effects of pectolinarigenin on the osteoclastogenesis TFs such as: NFATc1 and c-Fos, and further identify the relevant up-stream signals activity. We initially found pectolinarigenin inhibited receptor activator of nuclear factor-kappa B ligand (RANKL) induced osteoclast formation during the bone marrow-derived macrophages (BMMs) cultures, suggesting that this natural product could act on osteoclast precursors by inhibiting the down signaling cascades of RANKL signaling. Moreover, mechanistical investigation showed pectolinarigenin inhibits RANKL-mediated osteoclastogenesis by attenuating the nuclear factor of activated T cells cytoplasmic 1 (NFATc-1) and c-Fos following the Akt and mitogen activated protein kinases (MAPKs) signaling costimulatory. These findings identify that pectolinarigenin may act as an anti-resorption agent by blocking osteoclast activation.