Dihydroartemisin in inhibitions of RANKL-induced osteoclastogenesis in RAW264.7 cell line

Abstract

Objective To investigate the effects of dihydroartemisinin on RANKL-induced osteoclastogenesis. Methods Cell counting kit-8(CCK-8) was performed to exam the impact of dihydroartemisinin (1, 5, 10, 50, 100, 200 μmol/L) on viability of RAW264.7 cell line. RAW 264.7 cells were cultured with 50 ng/mL RANKL as well as dihydroartemisinin (5, 10, 50, 100 μmol/L) for 3 days to induce osteoclast formation, following with a TRAP staining. TRAP-positive cells with more than three nucleuses were counted. RAW264.7 cells were cultured with 50 ng/mL RANKL as well as 10 μmol/L and 100 μmol/L dihydroartemisinin for 24 h and total RNAs were extracted with Trizol reagent. The expressions of osteoclast-related genes were measured by real time-PCR, such as NFATc1, c-fos, Cathepsin K. Results Dihydroartemisinin (1, 5, 10, 50, 100 μmol/L) showed no significant cytotoxicity on RAW264.7 cell line for 24 h. Dihydroartemisinin (5, 10, 50, 100 μmol/L) inhibited RANKL-induced osteoclast formationin a dose-related manner. Dihydroartemisinincan (10, 100 μmol/L) down-regulated the expression of key osteoclast-forming gene NFATc1 and c-fos, with no significant influence on Cathepsin K. Conclusions Dihydroartemisinin is low cytotoxicity on RAW264.7 cell line. Dihydresoartemisinin can inhibit RANKL, induce osteoclastogenesis via down-regulating the expression of NFATc1 and c-fos. Dihydroartemisinin can be a potential pharmacological treatment for bone destruction disease. Key words: Artemisinin; Dihydroartemisinin; Receptor activator for nuclear factor-κ B ligand; Osteoclasts; RAW264.7 cell line