ABSTRACT
BACKGROUND:Recently, glucocorticoid-induced necrosis of femoral head has been much studied. However, the precise Wnt signaling pathway by which puerarin suppresses adipogenic differentiation of glucocorticoid-induced bone marrow mesenchymal stem cells adipogenic differentiation of glucocorticoid-induced bone marrow mesenchymal stem cells remains unconfirmed. OBJECTIVE:To investigate the expression of Wnt signaling pathway related genes and the key factor protein,β-catenin, during adipogenic differentiation of glucocorticoid-induced rat bone marrow mesenchymal stem cells treated by puerarin. METHODS:The third generation of bone marrow mesenchymal stem cells were cultured with Dulbecco’s modified Eagle’s medium containing blank serum (blank control group), dexamethasone (hormone group), dexamethasone with puerarin low dose group, the middle dose group and high dose group. After 6 days of culture, in the above five groups, the expressions of Wnt/β-catenin signaling pathway members, Wnt10b mRNA, GSK3βmRNA,β-catenin mRNA, were detected using RT-PCR assay, and the expression ofβ-catenin protein was detected using western blot assay. RESULTS AND CONCLUSION:Compared with the control group, the Wnt10b mRNA,β-catenin mRNA andβ-catenin protein expressions were significantly higher in puerarin groups, but GSK3βmRNA expression was significantly lower in the puerarin groups. These findings suggest that puerarin effects on inhibition of adipogenic differentiation of glucocorticoid-induced bone marrow mesenchymal stem cells probably are realized through the activation of Wnt/β-catenin signal pathway and regulation of the key factor Wnt10b mRNA, GSK3βmRNA,β-catenin mRNA, andβ-catenin protein expressions. The mechanism by which puerarin prevents glucocorticoid-induced necrosis of femoral head not only improves local microcirculation of the femoral head, but also relates to its inhibitory effects on adipogenic differentiation of glucocorticoid-induced bone marrow mesenchymal stem cells.