NFATc1过表达对唑来膦酸诱发的破骨细胞生成抑制的影响  

The effect of NFATc1 over-expression on zoledronate-induced inhibition of osteoclastogenesis

作　　者：张广峰 阴露佳 戚孟春 董伟 冯晓洁 温黎明 孙红 

机构地区：[1]华北理工大学口腔医学院口腔颌面外科教研室,河北唐山063000 [2]华北理工大学基础医学院病理教研室,河北唐山063000

出　　处：《广东医学》2017年第12期1795-1799,共5页Guangdong Medical Journal

基　　金：国家自然科学基金资助项目(编号:81270965);河北省高等学校科学技术研究重点项目(编号:ZD2015005);河北省自然科学基金资助项目(编号:2011401044)

摘　　要：目的研究活化T细胞核因子蛋白c1(NFATc1)过表达对唑来膦酸诱发的破骨细胞生成抑制的挽救效应。方法小鼠NFATc1重组慢病毒转染RAW264.7细胞,验证NFATc1蛋白表达。细胞分为A、B两组,分别转染空白载体和NFATc1重组载体。两组细胞均用50 ng/mL核因子κB受体激活蛋白配体(RANKL)诱导5 d,并在第2天加入1×10^(-6)mmol/mL唑来膦酸作用2 d。第6天收获细胞,检测破骨细胞的生成和功能及NFATc1、抗酒石酸酸性磷酸酶(TRAP)、酪氨酸激酶(c-Src)基因的表达情况。结果 NFATc1蛋白在细胞中均有表达。B组TRAP阳性破骨细胞数、牙本质吸收陷窝数目和面积显著高于A组(P<0.01)。B组NFATc1、TRAP、c-Src的蛋白水平显著高于A组(P<0.01);B组3个基因mRNA水平显著高于A组(P<0.01)。免疫荧光细胞化学检测也得到相似的结果。结论 NFATc1过表达对唑来膦酸诱发的破骨细胞生成及NFATc1、TRAP、c-Src基因表达的抑制具有挽救效应;NFATc1作为Ca^(2+)信号的关键分子,在唑来膦酸诱发的破骨细胞生成抑制中起着关键作用。Objective To investigate the rescue effect of nuclear factor of activated T cells type cl （NFATcl） over - expression on zoledronate - induced inhibition of osteoclastogenesis. Methods RAW264.7 cells were transfected with mouse recombinant NFATcl lentivirus, and protein expression of NFATc1 was verified. The ceils were divided into Group A and B, in which the cells were transfected with blank vector and recombinant NFATcl vector, respectively. The cells in both groups were treated with 50 ng/mL RANKL for 5 days and 1 × 10^- 6 mmol/mL zoledronate for 48 h from Day 2. The ceils were harvested on Day 6 and osteoclastogenesis, as well as gene expression of NFATcl, TRAP and c - Src were examined. Results NFATcl protein was successfully expressed in RAW264.7 cells. TRAP positive muhinuclear osteoclast count, and the number and size of absorption lacunae on dentin slices in Group B were all significantly higher than those in Group A （P 〈0. 01 ）. Protein levels of NFTAcl, TRAP and c - Src in Group B were also significantly higher than those in Group A （P 〈0. 01 ）. And mRNA levels of above three genes in Group B were significantly higher than those in Group A （ P 〈 0. 01 ）. Similar results were also achieved in immunofluorescent cytochemistry examination. Conclusion Over - expression of NFATcl can rescue zoledronate - induced inhibition of osteoclastogenesis and up - regulate gene expression of NFATcl, TRAP and c -Src. As a key transducer of Ca2＋ signal, NFATcl may play a key role in zoledr- onate - induced inhibition of osteoclastogenesis.

关 键 词：活化T细胞核因子蛋白c1 基因重组 唑来膦酸 破骨细胞 抗酒石酸酸性磷酸酶 

分 类 号：R68[医药卫生—骨科学]