补肾中药骨灵丸对成骨细胞矿化功能及骨桥蛋白基因表达的影响

目的探讨含补肾中药-骨灵丸大鼠血清对体外培养成骨细胞(OB)矿化功能及骨桥蛋白基因表达的影响。方法取SD乳鼠颅骨,采用改良胰酶-胶原酶消化法分离培养;根据血清药理学的方法制备含不同剂量(高、中、低剂量)骨灵丸血清,并以空白组(灌喂生理盐水)为阴性对照、雌二醇(E2,10-8 mol／L)组为阳性对照;将制备好的各组血清分别加入第二代OB的培养液中进行培养,采用Westem-blot及RT-PCR的方法观察补肾中药血清对OB骨桥蛋白及基因表达的影响,利用茜素红染色方法测定矿化结节形成的数量。结果含骨灵丸(中、高剂量组)血清均能上调OB骨桥蛋白及基因的表达,使钙结节的数目增加,与空白对照组相比较具有统计学意义(P<0．05);空白对照组及补肾中药低剂量组对骨桥蛋白表达及矿化结节的形成无影响。结论骨灵丸可上调骨桥蛋白基因的表达,这可能是骨灵丸促进钙结节形成、影响骨代谢的机理之一。

Effect of Kidney-tonifying Traditional Chinese Herbs on Osteopontin Gene Expression of Rat Osteoblast Cultured in Vitro

Objective To explore the effects of Chinese herbs on the gene expression of osteopontin(OPN) in rat cultured in vitro. Methods Osteoblasts were from the skull of newborn SD rats and treated with high, middle or low concentration of the Chinese herbs. RT-PCR and western blot were used to detect the expression of OPN. Results At both high and middle concentration of the Chinese herb could up regulate the levels of OPN RNA and protein. Conclusion Kidney-tonifying traditional Chinese herb can promote the function of bone formation. Increase in mineral nodes may be due to the up regulation of both OPN RNA and protein.