Research Paper Volume 12, Issue 21 pp 21706—21729

Puerarin alleviates osteoporosis in the ovariectomy-induced mice by suppressing osteoclastogenesis via inhibition of TRAF6/ROS-dependent MAPK/NF-κB signaling pathways

Figure 6. Puerarin reduces F-actin ring formation in the osteoclasts and suppresses osteoclastogenesis-related gene expression. (A) Representative images show the podosome belt formation in the osteoclasts formed from the RANKL and RANKL+ indicated concentrations of puerarin-treated BMMs. The cells were stained with antibodies against MMP9/NFATc1 (red), phalloidin for F-actin (green) and the nuclei were stained with DAPI (blue). (B, C) Quantitative analysis of the F-actin ring area in the osteoclasts from the RANKL and RANKL+ indicated concentrations of puerarin-treated BMMs. Note: n=3 per group, ** P <0.01 vs. the control group (without puerarin treatment). (D, E) Quantitative analysis of the numbers of nuclei per osteoclast in the control BMMs and BMMs induced the RANKL and RANKL+ indicated concentrations of puerarin-treated. Note: n=3 per group; ** P <0.01 vs. the control group (without puerarin treatment). (F–J) QRT-PCR analysis of the mRNA levels of osteoclastogenesis-related genes, NFATc1, MMP9, CTSK, Acp5, and c-Fos, in the control BMMs and BMMs induced with RANKL and RANKL+ indicated concentrations of puerarin. Note: n=3 per group; ##P <0.01 vs. the control group (without RANKL induce and puerarin treatment); ** P <0.01 vs. the RANKL-induced group (without puerarin treatment).