| Objective To investigate the effects of resveratrol (RSV) at different concentrations on osteoclast differentiation and the role of autophagy. Methods RAW264.7 cells were added with RANKL and RSV of different concentrations (0, 0.1, 0.5, 1, 5 and 10μmol/L). Cell viability was measured by CCK-8 at 12, 24, 48 and 72 hours after the intervention. Osteoclast differentiation was observed by TRAP staining. With or without autophagy inhibitor 3-Methyladenine, RT-PCR was used to detect the expression of osteoclast differentiation related markers including TRAP, MMP-9, CTSK and autophagy related markers LC3, Beclin-1 and P62. Western Blot was used to detect the expression of autophagy related proteins LC3II/I, Beclin-1 and P62. Results In the process of RANKL induced differentiation, the cell proliferation activity increased, and the cell proliferation activity first increased and then decreased with the addition of 0.1-10μmol/L RSV, reaching the maximum at 0.5μmol/L. RSV with 0.1μmol/L and 0.5μmol/L increased TRAP positive osteoclast number and mRNA expression of CTSK, MMP9, TRAP, LC3, P62 and Beclin-1, autophagy related protein LC3II/I and beclin-1 expression were also increased, and P62 expression was decreased. With the increase of RSV concentration 1~10μmol/L, the number of osteoclasts and the expression of related mRNA and protein LC3II/I and beclin-1 decreased, and the expression of protein P62 increased. 3-Methyladenin inhibited the expression of related mRNA and protein LC3II/I and beclin-1, and the expression of protein P62 was increased. Conclusion In the range of 0.1-10μmol/L resveratrol concentration, the level of osteoclast differentiation and autophagy first increased and then decreased. Inhibition of autophagy can inhibit the differentiation of osteoclasts. The effect of resveratrol on osteoclast differentiation may be partly mediated by autophagy. |