Effect Of Astragalus Polysaccharides On The Activities Of Osteoblast In High Glucose Conditions

OBJECTIVE:To evaluate the effects of different concentrations of Astragalus polysaccharides (APS) on the proliferation?differentiation and mineralization of the MC3T3-E1 cell. Understand the change of the gene expression and discuss the mechanism of APS effect on MC3T3-E1 under the condition of high-glucose.METHODS:The MC3T3-E1 cell was cultured in vitro with 100g/L fetal bovine serum and a-MEM, under the condition of 37? and 5% saturation level of CO2, change the medium after 2-3 days. The generation followed the cell fusion beyond 90%. Afer the High-glucose configurated, the 0.5?0.05?0.005mg injection level of APS was dissolved in 100ml High-glucose medium respectively, and allocation to the High-dose APS group (HAD,5 mg/ml)?the Medium-dose APS group (MDA,0.5mg/ml)? the Low-dose APS group (LDA,0.05 mg/ml). the control group was simple High-glucose group (HG)Observe the proliferation of different groups by MTT assay in 1?3?5 days respectively, the activity of differentiation by ALP Kit in 1?3?5?7 days respectively and the mineralization of MC3T3-E1 cell by Alizarin red dye staining in 14?21 days respectively. The expression of Runx-2?OPN?OCN were examined by RT-P CR in the 3?7 days; the cytoskeletonic systemic was observed by confocal laser scanning microscopy (CLSM) in the early of the experiment.RESULTS:The data shows that the low-dose APS (LDA) and the medium-dose APS (MDA) group was more benefit for the proliferation of MC3T3-E1 cell, but there was no significantly dofferent between the LDA group and the MDA group.The LDA group's and differentiation and mineralization of MC3T3-E1 cell was significant differences with the MDA and high-dose APS (HDA) group by the observation of ALP Kit and Alizarin red dye staining (P<0.05).Instead of the other groups, the LDA group increased the gene expressions of Runx-2, OPN and OCN more significant in a time-dependent manner (P<0.05). The 0.05 mg/ml APS group was also more well growth and extension than the other groups at each time point by CLSM, which is benefit for the adhesion function of MC3T3-E1 cell.CONCLUSIONS:1?The high glucose condition is adverse for the proliferation, differentiation and mineralization of MC3T3-E1 cell, decreased the expression of OPN, Runx-2 and OCN, the growth and extension of cytoskeletonic is inhibited.2?0.05mg/ml APS, instand of the other concentrations APS, is better for the proliferation, differentiation and mineralization of MC3T3-E1 cell.3?Under the conditions of high glucose,0.05mg/ml APS can changeover the damage to the MC3T3-E1 cell, through the well growth and extension of cytoskeletonic and the increasing expression of OPN, Runx-2 and OCN. There is also increased the activity of the proliferation, differentiation and mineralization of MC3T3-E1 cell...