Effect Of Metformin (MF) On The Proliferation And Bone Morphogenetic Protein-2(BMP-2) And Core Binding Facter A1(Cbfa1) MRNA Expression In Osteoblasts

ObjectiveTo study the effect of metformin on proliferation and bone morphogenetic protein-2(BMP-2) and core binding facter al (Cbfal) mRNA expression in primary cultured rat calvarial osteoblasts. And investigate the possible mechanism of action on osteoblasts.Methods1Primary calvarial osteoblasts were isolated, cultured, identified and observed their morphology.2The milk rat osteoblasts were cultured in vitro, cultured osteoblasts were treated with metformin in different concentrations (0,50,100,200,400μmol/l), the proliferation of the milk rat osteoblasts is assessed by MTT colorimetry after24hours.3The osteoblasts that were cultured in vitro were treated with different dose of metformin(0,50,100,200,400μmol/l).The expression of BMP-2and Cbfal mRNA in primary cultured rat osteoblasts were analyzed by real-time quantitative RT-PCR after24hours.4statistical analysis Data are presented as X±S. All statistical analysis is performed with windows SPSS17.0.The differents between the groups are determined by One-way ANOVE. The differences between random two groups are determined by LSD. Differences are considered significant at a value of P<0.05.Results1Most of the milk rat osteoblasts were cultured in vitro were elongated spindle-shaped, irregular polygon or triangular,ALP staining was positive, cytoplasm was black. Alizarin red staining (after adding glycerophosphate-β and ascorbic acid-L into the medium) was red nodules.2Proliferation of primary cultured rat osteoblasts in vitro was promoted by different concentrations of metformin(0,50,100,200,400μmol/l) after24hours, The OD is increased. The experimental groups and compared with the control groups were considered significant at a value of P(P<0.05).3The primary cultured rat osteoblasts in vitro was treated with different concentrations of metformin(0,50,100,200,400μmol/l) after24hours, real-time quantitative RT-PCR examii revealed:metformin promoted BMP-2and Cbfal mRNA expression, in concentration-depe manner, The experimental groups and compared with the control groups were considered significant at a value of P(P<0.05).Conclunsion1、metformin induces the proliferation of primary cultured milk rat osteoblasts2、metformin promotes bone formation by increasing the expression of BMP-2and Cbfal mRNA.