Effects of sarsasapogenin on the activity of osteoblasts and the differentiation and the function of osteoclasts / 中国药科大学学报

ABSTRACT

Aim: To observe the effects of sarsasapogenin ( SAR) on osteoblasts and osteoclasts cultured in vitro. Methods: Colonal murine calvarial osteoblast-like cell line MC3T3-E1 cells were cultured in vitro. MTT,p-nitropheneye phosphate and tinctorial method of alizarin Bordeaux were used to investigate the effects of SAR on the proliferation, ALP expression, and mineralization tuberculation of MC3T3-E1 cells. Mature osteoclasts were i-solated from the long bone of one-day rat. Meanwhile, marrow cells of mouse bone were cultured with induction of 1,25( OH)_2VitD_3. During the culturing of osteoclasts or marrow cells, SAR of different concentrations was added into the medium. The number of osteoclasts was recognized as tartrate resistant acid phosphatase( TRAP) ( +) multinucleate cells and the resorption lacuna on bone slice were examined with toluidine blue staining. Results: Comparing with the control group, SAR (0.01, 0. 1, 1μg/mL) significanthy increased the proliferation of MC3T3-E1 cells (P <0. 05, P <0. 01). There was no significant difference in the expression of ALP in early pro-liferating MC3T3-E1 cells exposed to SAR of 0.01,0. 1, 1μg/mL, but in the differentiation phase MC3T3-E1 cells, SAR improved ALP activity very significantly if compared with the control group, of which SAR of 1 μg/mL had the most promotion effect(P <0. 01). In addition, compared to the control group, there were, to various ex-tents, increased in the number of mineral nodes in MC3T3-E1 cells after 15day incubation with SAR of different conentrations. Furthermore, no obvious effects of 0.01-1μg/mL SAR on mature osteoclast were observed. But typical osteoclasts were formed when marrow cells were cultured with the induction of 1,25(OH)_2D_3 in medium for 7 days while little or no osteoclasts were induced from marrow cells in the presence of SAR. Conclusion: The results suggest that SAR can effectively promote the proliferation, differentiation and mineralization of osteoblasts cultured in vitro. Besides, SAR can inhibit the generation of osteoclasts from marrow cells.