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 7. Puerarin inhibits ROS by upregulating antioxidant enzymes in the RANKL-induced RAW264.7 cells. (A) Flow cytometry analysis of ROS levels using DCFH2-DA in the untreated, RANKL-treated and indicated concentrations of puerarin+RANKL-treated RAW264.7 cells. Note: n=3 per group; ##P <0.01 vs. the untreated group; ** P <0.01 vs. the RANKL-treated group. (B) Representative western blot images show the levels of NOX1/2/4 and TRAF6 proteins in the untreated, RANKL-induced and indicated concentrations of puerarin+RANKL treated RAW264.7 cells. (CF) The histogram shows the relative levels of NOX1/2/4 and TRAF6 proteins in the untreated, RANKL- and RANKL+ indicated concentrations of puerarin-treated RAW264.7 cells. Note: n=3 per group; NS: Not statistically significant, ##P <0.01 vs. the untreated group; * P <0.05, ** P <0.01 vs. the RANKL-induced group. (G) Representative western blot images show the levels of antioxidant enzymes, GSR, HO-1, and catalase in the untreated, RANKL- and RANKL+ indicated concentrations of puerarin-treated RAW264.7 cells. (HJ) The histogram shows the relative levels of GSR, HO-1 and catalase proteins in the untreated, RANKL- and RANKL+ indicated concentrations of puerarin-treated RAW264.7 cells. Note: n=3 per group; ##P <0.01 vs. the untreated group; ** P <0.01 vs. the RANKL-induced group.