阿仑膦酸盐对破骨细胞生成的抑制及钙离子激动剂的拮抗效应

Alendronate inhibits osteoclastogenesis and calcium ionophore antagonizes the effects of alendronate

目的:研究阿仑膦酸盐(ALN)对破骨细胞(OC)生成及骨吸收功能的影响,并探讨钙离子激动剂对ALN的拮抗效应。方法:用小鼠单核巨噬细胞RAW264.7诱导法培养OC。实验分为:A组(对照组)、B组(ALN组)、C组(ALN+Calciumlonophore组)、D组(Calcium lonophore组)。于处理第6天检测各组OC生成和骨吸收功能,以及NFATc1、c-Fos基因表达情况。结果:各组细胞均有TRAP阳性多核OC生成,并在牙本质磨片上形成吸收陷窝;但D组TRAP阳性多核细胞数目、吸收陷窝数目及陷窝面积最高,C组、A组次之,B组最差。Real-time PCR和Western blot检测表明,NFATc1、c-Fos表达D组最高,C组次之,B组最差;与A组比较差异均有显著性(P<0.01)。结论:ALN能抑制OC生成和骨吸收,下调相关基因NFATc1、c-Fos的表达;Ca2+激动剂Calcium lonophore对ALN引起的OC抑制具有拮抗效应,其机制与细胞内Ca2+浓度调节有关。

Objective： To study the effect of alendronate（ALN） on osteoclastogenesis and bone resorption function and the antagonistic effect of calcium lonophore（CL） against ALN.Methods： Osteoclasts（OCs） were cultured by induction of mouse monocyte-macrophage cell line-RAW264.7.The OCs were divided into 4 groups： control（A）,ALN-treated（B）,ALN＋CL-treated（C） and calcium lonophore-treated（D）.At day 6 of treatment,osteoclastogenesis and resorption function of OCs were examined by TRAP staining and SEM observation respectively.The expressions of NFATc1 and c-Fos genes and protein were detected by RT-PCR and Western-blot respectively.Results： TRAP positive multinuclear cells were observed and resorption lacunas were formed in all 4 groups.Highest number of TRAP positive multinuclear cells and largest size of resorption lacunaes were observed in group D,followed in turn by group C,A and B.The highest expressions of NFATc1 and c-Fos mRNA and protein were found in group D,followed by group C,A and B（A vs B,C or D,P0.01）.Conclusion： ALN can inhibit osteoclastogenesis and resorption function of OCs,down-regulate gene expression of NFATc1 and c-Fos in OCs.Calcium lonophore has an antagonistic effect against ALN and the effect is associated with the regulation of ion intracellular Ca2＋ level.