Abstract
Oxysterols have been shown to inhibit adipogenic differentiation of mesenchymal stem cells while inducing their osteogenic differentiation. However, the effects of oxysterols on the adipogenic differentiation of C3H10T1/2 mouse stem cells are poorly understood. This study was conducted to examine the effects of specific oxysterols on the expression of adipogenic transcripts and adipogenic differentiation of C3H10T1/2 mouse stem cells. Cells were treated with 10% Fetal Bovine Serum (control), 500nM Dexamethasone, 0.5mM 3‐Isobutyl‐1‐methlyxanthine, 20µg/ml Insulin (DMI) + 10µM PPARγ agonist Troglitazone (Tro) (DMITro) and DMITro + 10µM 20S hydroxycholesterol (20S), 25 hydroxycholesterol (25), 22R hydroxycholesterol (22R) or 22S hydroxycholesterol (22S) for six days. Gene expression was analysed by real‐time quantitative polymerase chain reaction and expression data analysed using the Statistical Analysis Software (SAS 9.2). Compared to the control, DMITro significantly increased the expression of peroxisome proliferator‐activated receptor γ (PPARγ), CCAAT/enhancer binding protein α (C/EBPα), fatty acid binding protein 2 (FABP2) and lipoprotein lipase (LPL). Compared to DMITro, treatment of cells with DMITro + 25 significantly decreased the expression of PPARγ, the master regulator of adipogenesis (P 蠄 0.05). Expression of C/EBPα was significantly decreased by DMITro + 25 and DMITro + 20S (P 蠄 0.05), while the expression of FABP2 was significantly decreased by DMITro + 25, DMITro + 20S and DMITro + 22R (P 蠄 0.05). Lastly, the expression of LPL was significantly decreased by DMITro + 25 and DMITro + 22R (P 蠄 0.05). In conclusion, the results showed that 25 hydroxycholesterol was the most potent in inhibiting the expression of key adipogenic transcripts and adipogenesis of C3H10T1/2 mouse stem cells suggesting its potential application in reducing adipogenesis and obesity.Key words: Oxysterols, C3H10T1/2 stem cells, differentiation, PPARγGrant Funding Source: Supported by Canadian natural science and engineering research council.
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