NFATc1过表达对唑来膦酸诱发的破骨细胞生成抑制的影响

发布时间：2019-07-05 16:19

【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(P0.01)。B组NFATc1、TRAP、c-Src的蛋白水平显著高于A组(P0.01);B组3个基因mRNA水平显著高于A组(P0.01)。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。
文内图片：

图片说明：Westernblot检测ＲAW264．7细胞内NFATc1蛋白表达2．2破骨细胞生成及骨吸收功能检测经TＲAP
[Abstract]:Objective to study the effect of overexpression of activated T nuclear factor protein C1 (NFATc1) on osteoclast formation induced by zoledronic acid. Methods RAW264.7 cells were transfected with mouse NFATc1 recombinant lentivirus and the expression of NFATc1 protein was verified. The cells were divided into two groups: group A and group B, and were transfected with blank vector and NFATc1 recombinant vector, respectively. Both groups were induced with 50 ng/mL nuclear factor kappa B receptor activating protein ligand (RANKL) for 5 days, and 1 脳 10 ~ (- 6) mmol/mL zoledronic acid was added on the 2nd day for 2 d. On the 6th day, the formation and function of osteoclasts and the expression of NFATc1, anti-tartrate acid phosphatase (TRAP), tyrosine kinase (c-Src) gene were detected. Results NFATc1 protein was expressed in all the cells. The number of TRAP positive osteoclasts, the number and area of dentin resorption lacunae in group B were significantly higher than those in group A (P 0.01, the protein level of NFATc1,TRAP,c-Src in group). B was significantly higher than that in group A (P 0.01), and the level of mRNA of three genes in group B was significantly higher than that in group A (P 0.01). Similar results were obtained by immunofluorescence cytochemistry. Conclusion NFATc1 overexpression has a rescue effect on osteoclast formation and NFATc1,TRAP,c-Src gene expression induced by zoledronic acid. NFATc1, as a key molecule of Ca~ (2) signal, plays a key role in osteoclast formation inhibition induced by zoledronic acid.
【作者单位】：华北理工大学口腔医学院口腔颌面外科教研室;华北理工大学基础医学院病理教研室;
【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)
【分类号】：R965

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