Type I Saikosaponins A and D Inhibit Osteoclastogenesis in Bone Marrow-Derived Macrophages and Osteolytic Activity of Metastatic Breast Cancer Cells

Figure 4

Effects of saikosaponins A and D on osteoclasts activity and the formation of resorption pits. BMMs (2 × 10⁶ cells/well) were seeded into a 6-well culture plate and incubated in a medium containing M-CSF (30 ng/mL), RANKL (100 ng/mL), and saikosaponin A or D for 5 days. Control cells were cultured in the presence of M-CSF alone. The culture medium was collected and concentrated. (a) The levels of MMP-2 and MMP-9 in the conditioned medium were determined by gelatin zymography. Relative activity was described as fold by densitometric analysis. (b) Cathepsin K level was detected with the Sensizym cathepsin K activity assay kit. (c) BMMs (5 × 10⁴ cells/well) were treated with M-CSF (30 ng/mL), RANKL (100 ng/mL), and the indicated concentrations of saikosaponin A (SSA) or D (SSD) on Biocoat Osteologic Multitest slides for 11 days after the induction of osteoclast formation. Control cells were treated with M-CSF alone. Cells were lysed and the pit areas were observed under phase-contrast microscopy. Data are expressed as the mean ± SE. ^# versus control and ^* and ^** versus RANKL-treated BMMs.

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