Abstract
Background: Adipocyte and osteoblast derive from the same mesenchimal progenitor. Age-related decrease in bone mass is accompanied by an increase in marrow adipose tissue. Vitamin D3 (VD3) inhibits adipogenesis in 3T3-L1 preadipocytes. Recently it has been demonstrated that alendronate (ALN) inhibits adipogenesis while promoting osteoblast differentiation of mesenchimal stem cells. Aim of the Study: To evaluate the role of ALN on adipocyte differentiation in vitro and the potential synergic role of VD3 co-treatment. Procedures: Murine 3T3-L1 and 3T3-F442A preadipocytes were routinely differentiated in presence of ALN and VD3 10-9 - 10-7 M for 7 days and then stained with Oil Red O. The effect of these treatments on mRNA expression of the main molecular markers of adipocyte differentiation (PPARγ and C/EBPα) and VD Receptor (VDR) were analyzed through RT-PCR. Results: Both ALN and VD3 showed a marked anti-adipogenic effect on 3T3-L1 cells. Co-incubation of ALN 10-8 M and VD3 10-9 M displayed no synergic effect on inhibition of adipogenesis. PPARγ mRNA expression was significantly reduced by ALN and VD3. mRNA expression of C/EBPα was reduced only by VD3 treatment. An increase in VDR mRNA expression of 3T3-L1 cells was observed with both ALN and VD3. On the contrary, 3T3-F442A cells, which are in a more advanced adipogenic differentiation stage compared to 3T3-L1, did not express detectable levels of VDR. Interestingly, adipose differentiation of 3T3-F442A was not affected by ALN nor VD3. These results suggest that VDR may represent the molecular target of the anti-adipogenic effect of ALN. Conclusion: VDR plays a critical role in mediating the anti-adipogenic effect of ALN. Further studies to clarify this mechanism are warranted.
Highlights
Osteoporosis and fragility fractures represent a significant challenge for Health Systems in the western world because of their increasing number, a condition directly related to the steady growth of the elderly population
Murine 3T3-L1 and 3T3-F442A preadipocytes were grown until confluence at 37 ̊C in Dulbecco’s modified Eagle’s medium (DMEM) (Invitrogen, Paisley, Scotland) containing 4.5 g/liter D-glucose, 10% fetal calf serum (FCS) (Invitrogen), 100 units/ml penicillin, and 100 μg/ ml streptomycin
10−8 M and Vitamin D3 (VD3) 10−9 M did not display any synergic effect in terms of inhibition of adipogenesis (Figure 2)
Summary
Osteoporosis and fragility fractures represent a significant challenge for Health Systems in the western world because of their increasing number, a condition directly related to the steady growth of the elderly population. It has been demonstrated that ALN inhibits adipogenesis while promoting osteoblast differentiation of mesenchimal stem cells (MSCs) [2]. It has been demonstrated that alendronate (ALN) inhibits adipogenesis while promoting osteoblast differentiation of mesenchimal stem cells. Procedures: Murine 3T3-L1 and 3T3-F442A preadipocytes were routinely differentiated in presence of ALN and VD3 10−9 10−7 M for 7 days and stained with Oil Red O The effect of these treatments on mRNA expression of the main molecular markers of adipocyte differentiation (PPARγ and C/EBPα) and VD Receptor (VDR) were analyzed through RTPCR. PPARγ mRNA expression was significantly reduced by ALN and VD3. mRNA expression of C/EBPα was reduced only by VD3
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