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目的观察淫羊藿提取液(epimedium extract,EE)和雌二醇(17β-Estradiol,E2)联合使用对体外培养大鼠成骨细胞增殖和分化的影响。方法用酶消化法分离新生SD大鼠颅盖骨成骨细胞。用基础培养基培养3 d后获得未分化成骨细胞(undifferenciation osteoblast,udOB),继续用条件培养基培养3 d定向诱导分化为成骨细胞(differenciation osteoblast,dOB),分别以低剂量(1 mg/L)、中剂量(5 mg/L)及高剂量(10 mg/L)EE与10μmol/LE2联合干预48 h,检测细胞增殖率、碱性磷酸酶(alkline phosphatase,AKP)比活性及骨连接素(osteonectin,ON)基因mRNA水平。结果对udOB,高剂量联用组的增殖率及各联用组的AKP活性显著高于其他组,但ON的mR-NA水平没有显著性差异;对dOB,高剂量联用组的增殖率也明显高于其他组,但各联用组的AKP活性较对照组降低,而ON的mRNA水平上调。结论 EE和E2联合使用,较低剂量EE即有协同促进成骨细胞增殖作用,但高剂量EE的协同促增殖效应更强,而协同促分化作用主要表现在成骨细胞的分化早期,对分化成熟的成骨细胞有促成骨作用。
Objective To observe the effect of combined use of epimedium extract (EE) and estradiol (17β-estradiol, E2) on the proliferation and differentiation of cultured rat osteoblasts. Methods The calvaria osteoblasts of newborn SD rats were isolated by enzyme digestion. Undifferentiated osteoblasts (udOB) were obtained after culturing with basal medium for 3 days and were further induced to differentiate into osteoblasts (dOB) with conditioned media for 3 days. The cells were treated with low dose (1 mg / L), middle dose (5 mg / L) and high dose (10 mg / L) of EE and 10μmol / LE2 for 48 h, the cell proliferation rate, the specific activity of alkaline phosphatase (AKP) Osteonectin (ON) gene mRNA level. RESULTS: The proliferation rate of udOB and high-dose combination group and the AKP activity of each combination group were significantly higher than those of the other groups, but the mR-NA level of ON had no significant difference. For the proliferation rate of dOB and high-dose combination group Significantly higher than other groups, but the combined group of AKP activity decreased compared with the control group, while ON mRNA levels were increased. Conclusions Combination of EE and E2 can promote the proliferation of osteoblasts at lower dose of EE, but synergistic effect of high dose of EE is more effective than synergistic effect of E2. The synergistic effect of synergistic differentiation is mainly in the early differentiation of osteoblasts, Mature osteoblasts promote bone formation.