红车轴草总异黄酮含药血清对MC3T3-E1细胞增殖、分化和矿化的影响

Effect of trifolium pretense-derived total isoflavones containing serum on the proliferation,differentiation,and mineralization of MC3T3-E1 cell

目的研究红车轴草总异黄酮含药血清对MC3T3-E1成骨样细胞增殖、分化和矿化的影响。方法大鼠灌胃红车轴草总异黄酮一定时间后,制备含药血清,并分别采用MTT法、碱性磷酸酶活性测定法、ELISA法和茜素红染色测定矿化结节的方法考察不同浓度红车轴草总异黄酮含药血清对MC3T3-E1成骨样细胞的增殖率、碱性磷酸酶活性、骨钙素生成以及矿化结节形成的影响。结果不同浓度的红车轴草总异黄酮含药血清都能显著促进MC3T3-E1成骨样细胞的增殖并显著提高MC3T3-E1成骨样细胞碱性磷酸酶的活性,且最佳的含药血清浓度为5%;5%的红车轴草总异黄酮含药血清处理MC3T3-E1成骨样细胞6 d和9 d可显著促进骨钙素的分泌;5%的红车轴草总异黄酮含药血清处理MC3T3-E1成骨样细胞6 d、12 d和18 d,形成的矿化结节均显著高于对照组。结论红车轴草总异黄酮含药血清可以显著促进MC3T3-E1成骨样细胞的增殖、碱性磷酸酶活性、骨钙素分泌和矿化,说明红车轴草总异黄酮含药血清确实可以促进MC3T3-E1成骨样细胞的分化成熟,该药物具有开发为抗骨质疏松症新药的潜力。

Objective To investigate the effect of trifolium pretense-derived total isoflavones containing serum on the proliferation,differentiation,and mineralization of MC3T3-E1 osteoblastic cells in vitro. Methods Rats were intragastrically administered with the t. pretense-derived total isoflavones to prepare drug containing serum. The proliferation of MC3T3-E1 osteoblastic cells was determined using MTT method. The activity of alkaline phosphatase and osteocalcin were determined using ELISA. The mineralized nodules were observed using Alizarin red staining method. Results Different concentrations of total isoflavones containing serum significantly promoted the proliferation and increased the activity of alkaline phosphatase of MC3T3-E1 osteoblastic cells. The best concentration of the serum was 5%. 5% of total isoflavones containing serum significantly promoted the secretion of osteocalcin in MC3T3-E1 osteoblastic cells at 6^th day and 9^th day,and promoted the formation of mineralized nodules in MC3T3-E1 osteoblastic cells at 6^th day,12^th,and 18^th day. Conclusion The t. pratense-derived total isoflavones containing serum significantly promotes the proliferation,the activity of alkaline phosphatase,the secretion of osteocalcin,and the mineralization of MC3T3-E1 osteoblastic cells,indicating that it can significantly promote the maturation of MC3T3-E1 osteoblastic cells. The total isoflavones has the potential to develop a new medicine for osteoporosis.