Effects Of Total Flavonoids From Radix Pueraria Obata And Puerarin On Osteogenic And Adipogenic Differentiation Of Bone Marrow Stromal Cells

The aim of this study was to evaluate the effects of total flavonoids of Radix Pueraria lobata (TF), Puerarin (PRN) and Daidzein (DZN) on the osteogenic and adipogenic differentiation of primary mouse bone marrow stromal cells (MSCs) and osteoblasts by using MTT, ALP activity assay, Oil red O staining, alizarin red S staining methods, and the related mechanism by using Western blot and real-time PCR.Our findings indicated that:1. On cellular level:TF, PRN and DZN can promote the osteogenic differentiation and inhibit the adipogenic differen-tiation of MSCs;2. On molecular level:TF, PRN and DZN can upregulate the gene and protein expressions of Runx2, the key transcription factor on osteogenic differentiation, and down-regualte the gene and protein expression of PPARy2, the key transcription factor on adipogenic differentiation;3. ERa is one of the possible targets of TF, PRN and DZN during osteogenic and adipogenic differentiation of MSCs.In conclusion, the results indicated that TF, PRN and DZN exhibit a dual regulatory role in the differentiation process of MSCs into the osteogenic and adipogenic lineages, causing a lineage shift toward osteoblasts. Therefore, the protective effects of TF, PRN and DZN on bone may be through increasing number of osteoblasts to directly enhance bone formation and decreasing the number of adipocytes in MSCs, and secreting less cytokines which may inhibit the formation and activation of osteoclasts. In addition, Runx2and ERa are the possible targets in above process.