| AimIn this study, we investigate the effects of the PPARy agonist and all trans retinoic acid on inducing osteogenic differentiation in mesenchymal stem cells.MethodsChemiluminescence, PCR, Western blot and reporter plasmid assay were used to demostrate the effects of Troglitazone and all-trans retinoic acid in inducing osteogenic differentiation of MEFs.1. Chemiluminescence analysis was taken for ALP activity of MEFs treated with Tro and/or ATRA.2. Alizarin Red S Staining was employed to determine the matrix mineralization of MEFs treated with Tro and/or ATRA.3. RT-PCR and Western-blot were used to detect the expressions of OPN and OC in MEFs treated with Tro and/or ATRA.4. Recombinant adenovirus was used to analyze the effect of different PPARy2expression level on the ALP activity induced Tro and/or ATRA.5. Luciferase reporter assay and Western-blot were used to detect the transcription activity of BMPR-Smad signaling pathway induced Tro and/or ATRA.6. Oil Red O Staining and Western-blot were introduced to demonstrate the adipogenic differentiation induced Tro and/or ATRA.Results1. We found that the treatments of Tro and/or ATRA can effectively promote the ALP activity in MEFs, an early osteogenic marker.2. The matrix mineralization of MEFs treated with Tro and ATRA was much more than that treated with either Tro or ATRA alone.3. There existed synergistic effect of Tro and ATRA on expressions of OPN and OC, the osteogenic differentiation markers.4. Exo genie overexpression of PPARy2in MEFs could promote the ALP activity induced by the combination of Tro and ATRA.5. The combination of Tro and ATRA promoted the transcription activity of BMPR-Smad Signaling.6. The adipogenic differentiation induced by Tro was inhibited by ATRA.ConclusionsThe combination of Tro and ATRA can promote the osteogenic differentiation of mesenchymal stem cell, which may be mediated by increasing the BMPR-Smad signaling and inhibiting the adipogenic differention induced by Tro at least. |
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