This study intends to explore miR-34a’s effect on BMSCs osteogenic differentiation of hyperlipidemia. 20 SD rats were equally assigned into control group (normal diet) and high-fat group (research diet). Cells were transfected with miR-34a mimic or negative control followed by meausring miR-34a, Dvl2, PPAR-γ, ALP, Runx2, and sp7, and ALP activity. The number of mineralized nodules, adipocytes, miR-34a and PPAR-γ expression in high-fat group were significantly increased and Dvl2, ALP, Runx2, and sp7 mRNA showed opposite profiles. Meanwhile, Runx2, ALP protein, cytoplasm and nuclear blue-black particles, Dvl2 protein and mRNA in miR-34a mimic group were significantly downregulated (P < 0.05). Additionally, the luciferase activity of wild-type plasmid+miR-34a mimic group was significantly lower than mutant group, indicating that miR-34a targets Dvl2. In conclusion, miR-34a inhibits the osteogenic differentiation of hyperlipidemia BMSCs by inhibiting the expression of Dvl2, Runx2 and ALP activity, indicating that it might be target in the hyperlipidemia treatment.