Abstract： Objective： To study the effect of Chinese herbal compounds of Bushen Jianpi Huoxue prescription on proliferation and differentiation of osteoblasts as well as expressions of bone morphogenetic protein-2(BMP-2) and Smad5. Methods：A total of 60 three-month- old female SD rats were randomly divided into the blank group，the Bushen Jianpi Huoxue prescription group and the alendronate sodium group，20 cases in each group. Osteoblasts were intervened after the preparation of the corresponding medicated serum：the proliferation activity of osteoblasts was detected by Cell Counting Kit-8 (CCK-8)，the differentiation ability of osteoblasts was detected by alkaline phosphatase(ALP)，the mRNA expressions of BMP-2 and Smad5 were detected by real- time quantitative PCR(RT- qPCR)，and the protein expressions of BMP-2 and Smad5 were detected by Western blot. Results：When the intervention was 12，24 and 48 hours，the cell activity in each group was increased gradually： after intervention for 48 hours， the cell activity in each group was decreased. During 48 hours of treatment， compared with those in the blank group and the alendronate sodium group， the cell activity in the Bushen Jianpi Huoxue prescription group was the highest(P＜0.05). The ALP staining of Bushen Jianpi Huoxue prescription group was the deepest，which was significantly deeper than those in the blank group and the alendronate sodium group. Compared with those in the blank group，the protein and mRNA expressions of BMP-2 and Smad5 in the Bushen Jianpi Huoxue prescription group were increased(P＜0.05)：the expression of BMP-2 mRNA in the alendronate sodium group was decreased(P＜0.05). Compared with those in the alendronate sodium group，the protein and mRNA expressions of BMP-2 and Smad5 in the Bushen Jianpi Huoxue prescription group were increased(P＜0.05). Conclusion：Chinese herbal compounds of Bushen Jianpi Huoxue prescription can promote the proliferation and differentiation of rat osteoblasts，and increase the mRNA and protein expressions of BMP-2 and Smad5.