Evidence-Based Complementary and Alternative Medicine / 2012 / Article / Fig 2

Research Article

Effects and Interaction of Icariin, Curculigoside, and Berberine in Er-Xian Decoction, a Traditional Chinese Medicinal Formula, on Osteoclastic Bone Resorption

Figure 2

Inhibitory effects of I, B, C, and their combination on osteoclasts. (a) The morphological change of osteoclast (×200). Osteoclasts in the coculture system of primary osteoblasts and bone marrow cells were cultured for 24 h and then treated with or without the I, B, C, and their combination for 10 days. (b) Osteoclasts were treated for 10 days and then stained for TRAP, and those possessing three or more nuclei were counted as osteoclast-like multinucleated osteoclasts under a microscope. (c) Osteoclasts in the coculture system of primary osteoblasts and bone marrow cells were cultured for 6 days and then treated with or without I, B, C, and their combination for 48 hours. TRAP activity was measured by p-nitrophenyl sodium phosphate assay. (d) Cells were treated as described in TRAP assay, and the activity of cathepsin K was measured by a fluorometer equipped with a 400 nm excitation filter and 505 nm emission filter. Data were presented as mean ± standard deviation. The experiments were repeated 3 times in five replicate samples ( ) , , compared with control. The number on the column is q value, indicating the interaction between I, B, and C.
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