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Puerarin has attracted increasing attention because of its beneficial effect on anti-osteoporosis,but the molecular mechanisms underlying its actions on osteoblasts are not fully understood.The current study was aimed to investigate the effect of puerarin on the proliferation of mouse MC3T3-E1 osteoblast-like cells in vitro and its underlying mechanisms.We found that puerarin significantly promoted the proliferation of osteoblasts,enhanced ALP activity and increased the expression of TGF-β1,Smad2,Smad3 and Runx2.microRNA(miRNA)expression profiles analysis showed that the expression of miR-204 and other 118 miRNAs changed after puerarin treatment.MiRNA target prediction programs also predicted that miR-204 may directly target Runx2.After treatment with puerarin for 48 h,the expression level of miR-204 was downregulated.Besides,miR-204 dramatically repressed the luciferase activity of wildtype Runx2 3′-UTR transfected cells,but not that of the mutant ones.Overexpression of miR-204 in osteoblasts significantly decreased the protein expression of Runx2,while inhibition of miR-204 enhanced Runx2s expression.In addtion,overexpression of miR-204 inhibited the cell proliferation and ALP activity of osteoblasts,while inhibition of miR-204 had the opposite effect.The results suggest that puerarin could promote MC3T3-E1 osteoblast-like cell proliferation,which might be related to the downregulation of miR-204 and the following activation of Runx2