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Aim:To test the hypothesis that genistein stimulates the osteoblastic differentia-tion through the p38 mitogen activated protein kinase (MAPK)-core-binding fac-tor 1 (Cbfa1) pathway.Methods:The activation of p38 MAPK was detected byWestern blotting.Alkaline phosphatase (ALP) activity and calcium depositionwere assessed for osteoblastic differentiation of bone marrow-derived mesenchy-mal stem cell (BMSC) cultures.The expression of Cbfa1 was analyzed at both themRNA and protein levels.The activity of Cbfa1 was detected by electrophoreticmobility shift assay.Bone sialoprotein (BSP),ALP,osteocalcin (OC),andosteopontin (OPN) gene transcription were also evaluated by either RT-PCR orNorthern blotting.Results:Genistein (0.01-1 μmol/L) dose dependently led tothe rapid and sustained activation of the p38 MAPK pathway in mouse BMSCcultures.Treatment with genistein (1 μmol/L) resulted in increased ALP activityand calcium deposition of BMSC cultures as a function of time.Genistein alsoenhanced Cbfa1 DNA binding activity and promoted the expressions of Cbfa1itself as well as several Cbfa1-regulated genes,including ALP,BSP,OC,and OPN.Concurrent treatment with p38 MAPK inhibitor (SB203580) diminished thegenistein-induced osteoblastic maturation and p38 MAPK-Cbfa1 activation inmouse BMSC cultures.Conclusion:These results indicated that genistein couldstimulate the osteoblastic differentiation of BMSC cultures through the p38MAPK-Cbfa1 pathway.
Aim: To test the hypothesis that genistein stimulates the osteoblastic differentiation-through the p38 mitogen activated protein kinase (MAPK) -core-binding fac-tor 1 (Cbfa1) pathway. Methods: The activation of p38 MAPK was detected by Western blotting. phosphatase (ALP) activity and calcium deposition were assessed for osteoblastic differentiation of bone marrow-derived mesenchy-mal stem cell (BMSC) cultures. The expression of Cbfa1 was analyzed at both RNA and protein levels. The activity of Cbfa1 was detected by electrophoretic mobility shift assay .Bone sialoprotein (BSP), ALP, osteocalcin (OC), andosteopontin (OPN) gene transcription were also evaluated by either RT-PCR or Northern blotting. Results: Genistein (0.01-1 μmol / L) dose dependently led tothe rapid and sustained activation of the p38 MAPK pathway in mouse BMSC treated with treatment of genistein (1 μmol / L) resulted in increased ALP activity and calcium deposition of BMSC cultures as a function of time. Genistein also enhanced C bfa1 DNA binding activity and promoted the expressions of Cbfa1 self as well as several Cbfa1-regulated genes, including ALP, BSP, OC, and OPN. Current treatment with p38 MAPK inhibitor (SB203580) diminished the genistein- induced osteoblastic maturation and p38 MAPK- Cbfa1 activation inmouse BMSC cultures.Conclusion: These results indicates that genistein couldstimulate the osteoblastic differentiation of BMSC cultures through the p38MAPK-Cbfa1 pathway.