TCM Curculigo Orchioides Enhances The Proliferation And Differentiation Of Osteoblastic Cells

ObjectiveTo study the effects of the Curculigo orchioides extracts or curculigoside on the proliferation and differentiation of MC3T3-E1cells in vitro.MethodsCell growth of MC3T3-E1cells was assayed by MTT method after cells were incubated with the Curculigo orchioides extracts or curculigoside at various concentrations (i.e.,0,1:1500,1:1000,1:500,1:250or0,10,20,50, and100μg/mL) in vitro. Meanwhile, vascular endothelial growth factor (VEGF), Fms-like tyrosine kinase-1(Flt-1), and bone morphogenetic protein-2(BMP-2) were also detected in cell supernatants and cell lysates by ELISA and western blot analysis, respectively.ResultsTreated with the low concentration of Curculigo orchioides extracts (1:1500group), no statistically significant differences in cell proliferation were observed vs. vehicle-treated control cells. Whereas, at the1:1000group, the1:500group and the1:250group Curculigo orchioides extracts significantly promotes the proliferation in MC3T3-E1cells in vitro (p<0.05). To be similar with, a significant increase in the proliferation of MC3T3-E1cells (i.e.,≥59.8%) was detected for cells treated with50or100μg/mL curculigoside (P<0.01in each case). However, at concentrations of curculigoside<50μg/mL, no statistically significant differences in cell proliferation were observed.In the ELISAs performed, treatment of MC3T3-E1cells with Curculigo orchioides extracts or curculigoside resulted in an increase in the levels of VEGF, Flt-1, and BMP-2detected. For example, levels of VEGF for cells treated with10,20,50, and100μg/mL curculigoside or1:1500,1:1000,1:500,1:250Curculigo orchioides extracts were significantly higher than the levels of VEGF associated with vehicle-treated control cells (P<0.05). The same concentrations of curculigoside or Curculigo orchioides extracts also increased the levels of Flt-1expressed (P<0.05). In addition, BMP-2levels also markedly increased after MC3T3-E1cells were treated with curculigoside or Curculigo orchioides extracts compared to the vehicle-treated control group (P<0.05). The same tendency was found in western blot analysis.ConclusionsCurculigoside or Curculigo orchioides extracts significantly enhances the proliferation of MC3T3-E1cells, and stimulates the production of VEGF, Flt-1, and BMP-2. The preliminary results of this study indicate that curculigoside or Curculigo orchioides extracts has a pivotal role in the process of osteoblastic bone formation, and therefore, may represent a novel target for the treatment of common metabolic bone diseases.