Effects of Thymoquinone on Adipocyte Differentiation in Human Adipose-Derived Stem Cells

Inhibition of adipocyte differentiation would be a key strategy to control obesity. Human adipose tissue-derived stem cells (ADSCs) are a promising tool for adipocyte differentiation research. Thymoquinone (TQ) as a potent antioxidant molecule may inhibit adipocyte differentiation. Herein, we aim to investigate the inhibitory effect of TQ on lipid differentiation in ADSCs. Quantification of cell surface markers was used by Flow-Cytometry and the effect of TQ on cell viability was assessed using the AlamarBlue test. ADSCs were subjected to induction of differentiation in the presence of non-cytotoxic concentrations of TQ (6.25, 12.5 and 25 μg/mL). Lipid accumulation was assessed using the Oil-Red O staining technique. Moreover, the expression of PPARγ (Peroxisome proliferator-activated receptor-γ) and FAS (Fatty Acid Synthetase) proteins was evaluated using Western blotting. Flow-cytometry demonstrated the expression of CD44, CD90, and CD73 as mesenchymal stem cell markers on the cell surface. At concentrations ≤100 μg/mL of TQ, no significant difference in cell viability was observed compared to the control. Lipid accumulation in ADSCs significantly decreased at 25 μg/mL (P < 0.001) and 12.5 μg/mL (P < 0.01) of TQ. The findings of the qualitative examination of Lipid Droplets also confirmed these results. Western-blot showed that TQ at 12.5 (p < 0.05) and 25 μg/mL (p < 0.01) reduced FAS/β-actin ratio compared to the positive group. TQ also decreased the expression of PPARγ at 6.25 μg/mL but not at higher concentrations. In conclusion, TQ may reduce differentiation of fat stem cells into fat cells through inhibition of the expression of PPARγ and FAS proteins and might be a potential anti-obesity compound.