龙牡壮骨颗粒对小鼠成骨细胞MC3T3-E1增殖、分化及矿化的影响

Effects of Longmuzhuanggu Granules on the proliferation,differentiation and mineralization of osteoblastic MC3T3-E1 cells

目的：研究龙牡壮骨颗粒对小鼠成骨细胞株MC3T3-E1增殖、分化和矿化的影响。方法：以小鼠成骨细胞株MC3T3-E1作为药物筛选的体外模型;采用终浓度分别为1,5,10,25,50μg/ml的龙牡壮骨颗粒溶液干预,MTT法检测细胞增殖率;同时检测药物作用不同时间后的碱性磷酸酶（ALP）活性;茜素红染色和Von Kossa钙化染色检测不同浓度药物对细胞钙化的影响。结果：龙牡壮骨颗粒溶液在5~50μg/ml范围内培养24和48 h可显著促进小鼠成骨细胞增殖;龙牡壮骨颗粒溶液在5~50μg/ml范围内培养48和72 h显著提高MC3T3-E1细胞ALP活性。龙牡壮骨颗粒浓度为10μg/ml和50μg/ml时,茜素红染色钙化结节显著增多,药物浓度在10~50μg/ml范围内,Von Kossa染色钙化结节显著增多。结论：龙牡壮骨颗粒可提高MC3T3-E1增殖、分化及矿化的能力。

Objective： To investigate the effects of Longmuzhuanggu Granules on the proliferation,differentiation and mineralization of osteoblastic MC3T3-E1 cells. Methods： The osteoblastic MC3T3-E1 cell line was used as a model for drug screening in vitro. Longmuzhuanggu solution was supplemented into the culture at doses of 1,5,10,25,50 μg /ml respectively. MTT method was adopted in proliferation analysis and the alkaline phosphatase（ ALP） activity in the cells was assayed after incubated for different times. The mineralization of cells was determined via Alizarin Red staining and Von Kossa staining,respectively. Results： The cell proliferation was promoted at concentrations from 5μg /mlto50 μg /mlfor 24 h and 48 h. Longmuzhuanggu Granules was effective in significantly increasing ALP activity of MC3T3-E1 cells cultured for48 h and 72 h at doses ranging from 5μg /mlto 50 μg /ml. Longmuzhuanggu Granules at concentrations of 10μg /mland 50μg /mlremarkably increased the bone nodule formation of cells by Alizarin Red staining. While the effective concentrations were from 10μg /mlto 50μg /ml for Von Kossa staining. Conclusion： Longmuzhuanggu Granules has effects on stimulating the proliferation,differentiation and mineralization of MC3T3-E1 cells.