Fig. 5
The ability of leptin synergistically with 25(OH)D3 promoted osteoblast differentiation of rBMMSCs was weakened by the PI3K/AKT signal pathway inhibitor. a The increased mRNA levels of ALP, COL1A1, RUNX2, OSX, and OSP induced by 25(OH)D3 combined with leptin were significantly decreased after the addition of LY294002 (20 μM) at 12 h; b the protein levels of RUNX2 and COL1A1 induced by leptin and 25(OH)D3 were significantly inhibited by the inhibitor LY294002 (20 μM) at day 3; c relative quantitative analysis of western blot analyses; d the synthesis of 1α,25(OH)2D3 by rBMMSCs with ODM at 24 h; e results of ALP staining; f the relative expression level of ALP activity; g results of ARS staining; h the relative expression level of ARS. (Scale bar = 200 μm, **P < 0.01, ***P < 0.001 in comparison with the control group; #P < 0.05, ##P < 0.01, ###P < 0.001 in comparison with leptin + 25(OH)D3 group.)