双膦酸盐对破骨细胞分化及抗酒石酸酸性磷酸酶的影响  

Effect of bisphosphonate on osteoclast differentiation and tartrate-resistant acid phosphatase

作　　者：董伟[1] 冯晓洁[1] 梁永强[1] 邓久鹏[1] 温黎明[1] 戚孟春[1] 

机构地区：[1]河北联合大学口腔医学院,河北省唐山市063000

出　　处：《中国组织工程研究》2014年第38期6069-6073,共5页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金(81270965);河北省自然科学基金(C2011401044)~~

摘　　要：背景:抗酒石酸酸性磷酸酶是破骨细胞分化及骨吸收功能的特异性标志酶,是破骨细胞分化成熟的标志。目的:观察双膦酸盐对破骨细胞分化及骨吸收功能相关因子抗酒石酸酸性磷酸酶的影响。方法:小鼠单核巨噬细胞RAW264.7诱导培养破骨细胞。实验分2组:对照组开始时加入质量浓度100μg/L核因子kB受体活化因子配体进行诱导至收获细胞,双膦酸盐组在对照组的基础上加入10-7 mol/L阿仑膦酸盐处理至收获细胞。培养第7天检测各组破骨细胞生成和骨吸收功能,免疫荧光检测两组抗酒石酸酸性磷酸酶表达的差异,Western blot检测抗酒石酸酸性磷酸酶蛋白表达情况。结果与结论:各组细胞均有抗酒石酸酸性磷酸酶阳性多核破骨细胞生成,并在牙本质磨片上形成吸收陷窝;但对照组抗酒石酸酸性磷酸酶阳性多核细胞数目、吸收陷窝数目及陷窝面积均大于双膦酸盐组(P<0.01)。免疫荧光检测显示,对照组抗酒石酸酸性磷酸酶表达均强于双膦酸盐组(P<0.01)。Western blot检测显示,双膦酸盐组抗酒石酸酸性磷酸酶蛋白的表达低于对照组(P<0.01)。说明双膦酸盐通过抑制抗酒石酸酸性磷酸酶蛋白的表达,阻碍破骨细胞分化生成及骨吸收功能。BACKGROUND：Tartrate-resistant acid phosphatase is a specific marker for osteoclast differentiation and bone resorption, which is a sign of osteoclast maturity. 〈br〉 OBJECTIVE：To study the effect of alendronate on tartrate-resistant acid phosphatase related to osteoclast differentiation and bone resorption. 〈br〉 METHODOsteoclasts were cultured by mouse monocyte-macrophage cellline-RAW264.7. The cells were divided into two groupcontrol group, treated with 100μg/L receptor activator of nuclear factorκB ligand factor;alendronate group, treated with 100μg/L receptor activator of nuclear factorκB ligand factor＋10-7 mol/L alendronate. Osteoclastogenesis and resorption function of osteoclasts were examined at 7 days of culture. Gene expression of tartrate-resistant acid phosphatase was detected by immunofluorescence method. Western blot assay was used to detect protein expression of tartrate-resistant acid phosphatase. 〈br〉 RESULTS AND CONCLUSION：Tartrate-resistant acid phosphatase positive multinuclear cells were observed and resorption lacunae formed in two groups. Control group showed the higher number of tartrate-resistant acid phosphatase positive multinuclear cells and larger size of resorption lacunae than the alendronate group （P＆lt;0.01）. Immunofluorescence showed expression of tartrate-resistant acid phosphatase was higher in the control group than the alendronate group （P＆lt;0.01）;furthermore, the protein expression of tartrate-resistant acid phosphatase was also lower in the alendronate group than the control group （P＆lt;0.01）. These findings indicate that bisphosphonates could strongly inhibit osteoclastogenesis and its resorption function by inhibiting protein expression of tartrate-resistant acid phosphatase.

关 键 词：组织工程 二膦酸盐类 破骨细胞 骨质疏松 组织构建 骨组织构建 双膦酸盐 抗酒石酸酸性磷酸酶 免疫印迹法 骨吸收陷窝 免疫荧光化学检测 扫描电镜 牙本质磨片 核因子kB受体活化因子配体 国家自然科学基金 

分 类 号：R318[医药卫生—生物医学工程]