Ligustrum japonicum Thunb. Fruits Exert Antiosteoporotic Properties in Bone Marrow-Derived Mesenchymal Stromal Cells via Regulation of Adipocyte and Osteoblast Differentiation

Figure 4

Effects of solvent fractions (H₂O, 85% aq. MeOH, n-BuOH, and n-hexane) on adipogenic and osteogenic differentiation of hBM-MSCs. (a) hBM-MSCs were seeded in 6-well plates and osteo-induced with or without fractions (10 and 50 μg/ml). Cellular ALP activity of osteo-induced hBM-MSCs was measured with a spectrophotometric enzymatic activity assay at day 7 of differentiation. (b) Adipo-induced hBM-MSCs in 6-well plates with or without fractions (10 and 50 μg/ml) were stained with Oil Red O at day 14 of differentiation. Images of cells show stained lipid droplets (left panel). Lipid accumulation levels were calculated by the colorimetric quantification of the dye removed from the wells (right panel) and given as percentage of the untreated differentiated control group. (c) Adipogenic and (d) osteogenic protein levels were analyzed with Western blotting. Western blot bands were densitometrically quantified and plotted as relative percentage of the untreated differentiated control group. β-Actin was used as internal loading control. and vs. the untreated differentiated control group. Scale bar: 50 μm.

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