Modulatory Effect of 1,25-Dihydroxyvitamin D3 on IL1β-Induced RANKL, OPG, TNFα, and IL-6 Expression in Human Rheumatoid Synoviocyte MH7A
Figure 3
Effect of 1,25(OH)2D3 on osteoclastogenesis in the presence of IL1β in osteoclast precursors RAW264.7 cell line. RAW264.7 were treated with M-CSF (20 ng/mL) and RANKL (50 ng/mL) for 3 days. At 3 days pretreatment, cells were washed three times with PBS and were then incubated with IL1β (20 ng/mL) in the presence or absence of 1,25(OH)2D3 (10 nM) or 1,25(OH)2D3 alone for 5 days. TRAP positive cells with numerous (>3) unstained nuclei were considered as mature osteoclasts. RAW264.7 treated with medium as negative control was shown in (a). TRAP positive cells in unwashed RAW264.7 treated with RANKL continually were shown in (b) and in washed RAW264.7 cells treated with medium were shown in (c). After washing, TRAP positive cells in RAW264.7 incubated with IL1β, 1,25(OH)2D3 or IL1β plus 1,25(OH)2D3 were shown in ((d)–(f)). TRAP positive cells number in different groups were shown in (g) (*). The diagram of treatment process in RAW264.7 was shown in (h).
