“更年春”方对大鼠成骨细胞增殖与分化及矿化的影响

Effects of "Gengnianchun Decoction" on Osteoblasts Proliferation, Differentiation and Mineralization in Rats

目的观察中药复方“更年春”药物血清对新生大鼠成骨细胞增殖、分化和矿化的影响。方法用酶消化法分离培养新生大鼠颅盖骨成骨细胞,加入含不同药物浓度的更年春药物血清条件培养液培养,生理盐水作为阴性对照组,尼尔雌醇作为阳性对照药物。MTT法测定细胞增殖率,对硝基苯酚法(PNPP)检测碱性磷酸酶(ALP)活性以观察成骨细胞活性,茜素红染色观察矿化结节个数。结果与生理盐水组相比,更年春组成骨细胞MTT值(OD570)及OD405值明显增加(P均<0.01),更年春高剂量组矿化结节数显著高于生理盐水组(P<0.01)。与雌激素组相比,中药高剂量组成骨细胞的OD570值(P<0.01)、OD405值(P<0.05)及矿化结节数均增高(P<0.05)。结论更年春在体外对成骨细胞的增殖、分化和矿化有明显促进作用,更年春高剂量明显优于雌激素,提示更年春防治骨质疏松的作用可能与其促进成骨细胞活性有关。

Objective To explore the effects of ＂Gengnianchun Decoction ＂on the activities of osteoblasts in vitro. Methods The osteoblasts were isolated from newborn rat calvaria by enzyme digestion, treated with different culture media. Different concentrations of Chinese traditional medicine （GNC） ： high, moderate and low level groups, nilestfiol （E group ）, saline （N group ）and sham（S group）. Microculture tetrozolium, p-nitrophenyl phosphate, alizarin red were used to observe the proliferation, activity of alkaline phosphatase （ALP） and the number of mineral nodes of osteoblasts cultured in vitro. Results The OD570 and OD405 of osteoblasts in different GNC groups were higher than those of N group（P〈 0.01）. Compared with E group, the OD570 and OD405 of H group increased obviously（P〈 0.01, P 〈 0.05） H and E groups had many more mineralization nodes than those of N group （P 〈 0.01 and P 〈 0.05 ,respectively）. The mineralization nodes of H group were more than that of E group （P 〈 0.05）. Conclusion GNC can improve the proliferation, differentiation and mineralization of osteoblasts cultured in vitro. The H group is better than E group; the mechanism of GNC preventing osteoporosis may be related with its stimulating osteoblasts activities.