Genistein stimulates osteoblastic differentiation via p38 MAPK-Cbfa1 pathway in bone marrow culture

Abstract

Aim: To test the hypothesis that genistein stimulates the osteoblastic differentiation through the p38 mitogen activated protein kinase (MAPK)-core-binding factor 1 (Cbfa1) pathway. Methods: The activation of p38 MAPK was detected by Western blotting. Alkaline phosphatase (ALP) activity and calcium deposition were assessed for osteoblastic differentiation of bone marrow-derived mesenchymal stem cell (BMSC) cultures. The expression of Cbfa1 was analyzed at both the mRNA and protein levels. The activity of Cbfa1 was detected by electrophoretic mobility shift assay. Bone sialoprotein (BSP), ALP, osteocalcin (OC), and osteopontin (OPN) gene transcription were also evaluated by either RT-PCR or Northern blotting. Results: Genistein (0.01-1 µmol/L) dose dependently led to the rapid and sustained activation of the p38 MAPK pathway in mouse BMSC cultures. Treatment with genistein (1 µmol/L) resulted in increased ALP activity and calcium deposition of BMSC cultures as a function of time. Genistein also enhanced Cbfa1 DNA binding activity and promoted the expressions of Cbfa1 itself as well as several Cbfa1-regulated genes, including ALP, BSP, OC, and OPN. Concurrent treatment with p38 MAPK inhibitor (SB203580) diminished the genistein-induced osteoblastic maturation and p38 MAPK-Cbfa1 activation in mouse BMSC cultures. Conclusion: These results indicated that genistein could stimulate the osteoblastic differentiation of BMSC cultures through the p38 MAPK-Cbfa1 pathway.

Role of Phytoestrogen Ferutinin in Preventing/Recovering Bone Loss: Results from Experimental Ovariectomized Rat Models

“…Regarding the bone turnover-related serum levels, the recorded values of alkaline phosphatase (the most widely recognized biochemical marker for osteoblastic activity - Evans et al, 1990;Nian et al, 2006) suggest that the process of osteogenesis should be triggered in F-OVX group, because ALP value in F-OVX is higher with respect to the other groups. A positive effect on osteoblast activity in vitro by other phytoestrogens, like genistein, has already been published (Liao et al, 2007;Pan et al, 2005). As far as ferutinin side effects is concerned on the organs commonly targeted by estrogens, the apparent above cited antiapoptotic effect on endometrial epithelia is in line with observations previously recorded for genistein by other authors that administered such phytoestrogen to ovariectomized mice (Eason et al, 2005;Garcìa-Pérez et al, 2006).…”

.BETA.-Ecdysterone Induces Osteogenic Differentiation in Mouse Mesenchymal Stem Cells and Relieves Osteoporosis

“…Increasing data show that many chemical components isolated from herbal medicines with structures similar to that of estrogen have functions similar to those of estrogen. [21][22][23] Our findings provided the first evidence that b-ecdysterone isolated from traditional Chinese medicine could induce the osteogenic differentiation of MSCs and did not have obvious effects on adipogenic differentiation.…”

Extra-cellular matrix suppresses expression of the apoptosis mediator Fas by epigenetic DNA methylation

“…Treatment with inhibitors of signal transduction was performed with the focal adhesion kinase inhibitor (FAK) genistein [16] (10 lM, Sigma), the selective MAP-/ERkinase inhibitor PD98059 [17] (10 lM, Promega) and the AP1 inhibitor resveratrol [18] (40 lM, Sigma). In these experiments, inhibitors were added to the cells 30 min before seeding.…”