阿仑膦酸钠、氟化钠联合作用对体外培养大鼠成骨细胞的影响

Effect of Alendronate Combined with Sodium Fluoride on Osteoblasts from Rats in Vitro

目的探讨阿仑膦酸钠联合氟化钠对体外培养大鼠成骨细胞的作用。方法采用酶消化法分离SD大鼠颅盖骨中成骨细胞,原代培养。分别给予不同浓度的阿仑膦酸钠和氟化钠分组刺激,观察药物对成骨细胞增殖和碱性磷酸酶活性的影响,并用Western Blot法测定骨保护素(osteoprotegerin,OPG)/核因子-B受体活化因子配体(receptor activator for nuclear factor-B ligand,RANKL)蛋白表达。结果阿仑膦酸钠最佳单药浓度为10^-6 mmol/L,氟化钠的最佳单药浓度为10^-8 mmol/L,两种药物最佳浓度联合应用更能促进成骨细胞增殖,碱性磷酸酶(alkaline phosphatase,ALP)活性更强,OPG表达升高、RANKL表达下调,与对照组相比,差异有统计学意义。高浓度氟化钠(10^-3 mmol/L、10^-4 mmol/L)对成骨细胞有抑制作用。结论阿仑膦酸钠、氟化钠低浓度时均促进成骨细胞增殖和分化,二者联合有协同作用。高浓度氟化钠对成骨细胞有抑制作用。

Objective To study the effect of alendronate combined with sodium fluoride on osteoblasts in vitro.Methods To observe the proliferation and alkaline phosphatase(ALP)activity,the osteoblasts primarily cultured in vitro were isolated from the skull of rats by trypsin digestion method,andstimulated by alendronate and sodium fluoride at different concentrations.The expression of OPG/RANKL was examined by Western Blotting.Results There was the most significant promotion in osteoblast differentiation and proliferation when the osteoblasts were cultured at the concentration of alendronate of 10^-6 mmol/L and sodium fluoride of 10^-8 mmol/L.Compared with the control group,the osteoblastsincombined therapy group had significantly promotion in proliferation and ALP activity.The expression of OPG was increased,and the expression of RANKL was decreased.There were statistically significantdifferences in proliferation,ALP activity and expression of OPG/RANKL.An inhibitory effect was observed at high concentrations of sodium fluoride(10^-3 mmol/L、10^-4 mmol/L).Conclusion Both alendronate and sodium fluoride can promote osteoblast proliferation and differentiation,and there is synergistic effect when they combined.Sodium fluoride at high concentrations had an inhibitory effect on osteoblasts.