Evidence-Based Complementary and Alternative Medicine / 2017 / Article / Fig 4

Research Article

Liensinine and Nuciferine, Bioactive Components of Nelumbo nucifera, Inhibit the Growth of Breast Cancer Cells and Breast Cancer-Associated Bone Loss

Figure 4

Liensinine and nuciferine inhibited RANKL-induced osteoclastogenesis and osteoclast-mediated bone resorption. (a) BMMs were incubated in 10% FBS-α-MEM containing M-CSF (30 ng/ml) and liensinine or nuciferine at various concentrations for 5 days. Cell viability was determined by an MTT assay. Data are expressed as the means ± SE. and versus BMMs without test compounds. (b) BMMs were treated with liensinine and nuciferine at the various concentrations in a presence of M-CSF and RANKL. TRAP-positive cells with >3 nuclei were counted as mature osteoclasts (100x). (c–e) BMMs were incubated in 10% FBS- α-MEM containing M-CSF (30 ng/ml) and RANKL (100 ng/ml) on calcium phosphate-coated plates for 5 days and then treated with liensinine or nuciferine for an additional 2 days. (c) After cell lysis, the pit area was observed under light microscopy (×100). (d) Cathepsin K levels in the collected culture media were measured using the SensiZyme Cathepsin K Activity Kit. (e) Gelatinolytic activities of MMPs in the collected culture media were detected by zymography and visualized as clear bands against the blue background. Data are expressed as the means ± SE. versus BMMs without RANKL. and versus BMMs with RANKL alone.