中国组织工程研究

中国组织工程研究 ›› 2014, Vol. 18 ›› Issue (33): 5293-5298.doi: 10.3969/j.issn.2095-4344.2014.33.008

• 骨组织构建 bone tissue construction • 上一篇    下一篇

双膦酸盐对破骨细胞分化中组织蛋白酶K及骨吸收功能的影响

董伟，冯晓洁，梁永强，彭宏峰，邓久鹏，温黎明，戚孟春   

  1. 河北联合大学口腔医学院，河北省唐山市  063000
  • 出版日期:2014-08-13 发布日期:2014-08-13
  • 通讯作者: 戚孟春，博士， 教授，硕士生导师，河北联合大学口腔医学院，河北省唐山市 063000
  • 作者简介:董伟，男，1981年生，辽宁省辽中县人，汉族， 2011年河北联合大学口腔医学院毕业，硕士，讲师，主要从事骨组织工程研究工作。
  • 基金资助:

    国家自然科学基金(81270965)；河北省2013年医学科学研究重点课题计划(20130056)

Bisphosphonate effects on capthesin K and bone resorption function during osteoclast differentiation  

Dong Wei, Feng Xiao-jie, Liang Yong-qiang, Peng Hong-feng, Deng Jiu-peng, Wen Li-ming, Qi Meng-chun   

  1. School of Stomatology, Hebei United University, Tangshan 063000, Hebei Province, China
  • Online:2014-08-13 Published:2014-08-13
  • Contact: Qi Meng-chun, M.D., Professor, Master’s supervisor, School of Stomatology, Hebei United University, Tangshan 063000, Hebei Province, China
  • About author:Dong Wei, Master, Lecturer, School of Stomatology, Hebei United University, Tangshan 063000, Hebei Province, China
  • Supported by:

    the National Natural Science Foundation of China, No. 81270965; the Medical Research Plan of Hebei Province in 2013, No. 20130056

PDF

458

被引次数

17

摘要：

背景：有研究表明双膦酸盐可抑制破骨细胞的骨吸收功能，但对其骨吸收功能关键细胞因子组织蛋白酶K是否产生作用，至今少有报道。
目的：观察双膦酸盐对破骨细胞分化中组织蛋白酶K及骨吸收功能影响。
方法：用小鼠单核巨噬细胞RAW264.7诱导培养破骨细胞。实验分2组：对照组加入质量浓度100 μg/L核因子κB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，培养72 h免疫荧光检测两组组织蛋白酶K表达差异，Western blot检测组织蛋白酶K蛋白表达情况。
结果与结论：两组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)。免疫荧光检测组织蛋白酶K表达对照组强于双膦酸盐组(P < 0.01)；Western blot检测组织蛋白酶K表达双膦酸盐组低于对照组(P < 0.01)。结果证实，双膦酸盐通过抑制组织蛋白酶K因子的表达，阻碍破骨细胞的骨吸收功能。



中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程


全文链接：

关键词: 组织构建, 骨组织工程, 双膦酸盐, 破骨细胞, 抗酒石酸酸性磷酸酶, 组织蛋白酶K, 骨吸收陷窝, 免疫印迹, 免疫荧光化学, 骨质疏松, 扫描电镜, 牙本质磨片, 国家自然科学基金

Abstract:

BACKGROUND: Studies have shown that bisphosphonates inhibit osteoclast resorption, but whether cathepsin K, a key cytokine of bone resorption, plays an effect has rarely been reported.
OBJECTIVE: To study the effect of bisphosphonate on capthesin K and bone resorption function during osteoclast differentiation.
METHODS: Osteoclasts were cultured by mouse monocyte-macrophage cell line-RAW264.7. The cells were divided into two groups: control group, treated with 100 μg/L receptor activator of nuclear factor κB ligand factor; alendronate group, treated with 100 μg/L receptor activator of nuclear factor κB ligand factor+10-7 mol/L alendronate. Osteoclastogenesis and resorption function of osteoclasts were examined at 7 days of culture and gene expression of capthesin K was detected by immunofluorescence method at 72 hours of culture. Western blot assay was used to detect capthesin K protein expression at 72 hours of culture.
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 < 0.01). Immunofluorescence showed expression of capthesin K was higher in the control group than the 
alendronate group (P < 0.01); furthermore, the protein expression of capthesin K was also lower in the alendronate group than the control group (P < 0.01). These findings indicate that bisphosphonates could strongly inhibit osteoclastogenesis and its resorption function by inhibiting gene expression of capthesin K.



中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程


全文链接：

Key words: tissue engineering, diphosphonates, osteoclasts, osteoporosis

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