Title: Effects of isopsoralen on the osteogenic and adipogenic differentiation of bone marrow mesenchymal stem cells in mice
Abstract: Objective
To explore the effects of isopsoralen on the osteogenic and adipogenic differentiation of bone marrow mesenchymal stem cells (BMSCs) in mice and their mechanisms.
Methods
BMSCs were isolated from 18 2-month old female C57/BL6 mice that were specific pathogen free. Induced osteoblasts and fat cells were established and identified through cell culture, cell proliferation assay, alkaline phosphatase (ALP) staining and Oil Red O staining. After 14 days' induction, the core binding protein 2 (RUNX2), osteocalcin (OCN), a peroxisome proliferator activated receptor gamma (PPAR-γ) and CCAAT enhancer binding protein beta (C/EBP-β) were detected to determine the role of isopsoralen in groups of isopsoralen concentrations of 0,5,10 and 20 μmol/L. Western blotting was used to detect the expression of P-S6 (S235/236) and 4E/BP1 (Thr37/46) in the signal pathways for osteogenic induction and adipogenic induction of BMSCs.
Results
After osteogenic induction for 14 days: the positive expression of ALP in BMSCs in the 10 and 20 μmol/L groups were increased compared to the O and 5 μmol/L groups, with the highest in the 20 μmol/L group; the expression of RUNX2 and OCN in the 5,10 and 20 μmol/L groups was significantly increased compared to the 0 μmol/L group, with the highest in the 20 μmol/L group ( P < 0.05) . After adipogenic induction for 14 days: the isopsoralen in the 5,10 and 20 μmol/L groups significantly inhibited the expression of PPAR-γ and C/EBP-β in BMSCs, with the most in the 20 μmol/L group (P < 0.05) . After osteogenic induction for 14 days: the expression of P-S6 (S235/236) was obviously decreased, with the lowest in the 20 μmol/L group; the expression of 4E/BP1 (Thr37/46) was obviously increased, with the lowest in the 0 μmol/L group. After adipogenic induction for 14 days: the expression of P-S6 (S235/236) was obviously increased, with the lowest in the 0 μmol/L group; the expression of 4E/ BP1 (Thr37/46) was obviously decreased, with the lowest in the 20 μmol/L group.
Conclusions
Isopsoralen may enhance osteogenesis of BMSCs and inhibit BMSCs from differentiation into fat cells. The present experiment has also confirmed that isopsoralen inhibits the adipogenic differentiation of BMSCs by inhibiting the activity of mTORCl signaling pathway.
Key words:
Bone marrow cells; Osteoblasts; Adipogenesis; Isopsoralen; Osteogenic
Publication Year: 2015
Publication Date: 2015-12-15
Language: en
Type: article
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