摘要
目的研究双膦酸盐(ALN)对破骨细胞(OC)分化过程中关键因子NFATc1及其信号分子p-NF-κB、p-c-Jun的影响。方法采用小鼠单核巨噬细胞RAW264.7诱导培养OC,细胞培养于培养皿,分A组(对照组)、B组(ALN组)。培养第2d采用Western blotting检测NFATc1、p-NF-κB、p-c-Jun基因表达变化情况,培养第4d采用免疫荧光方法检测NFATc1表达,培养第7d检测各组OC生成及数目。采用牙本质磨片接种RAW264.7细胞,分组及处理方法同上述培养皿培养过程,第9d检测骨吸收功能。结果两组细胞均有TRAP阳性多核OC生成,并在牙本质磨片上形成吸收陷窝;但A组TRAP阳性多核细胞数目、吸收陷窝数目及陷窝面积均大于B组。免疫荧光检测NFATc1表达A组高于B组;Western blotting检测NFATc1、p-NF-κB、p-c-Jun表达B组较A组低,两组有显著性差异(P<0.01)。结论双膦酸盐通过下调p-NF-κB、p-c-Jun的表达,最终抑制NFATc1的表达,从而抑制OC生成和骨吸收功能。
Objective To study the effect of alendronate （ALN） on NFATcl and correlated signaling factors p-NF-KB and p-c-Jun in osteoclast differentiation. Methods Osteoclasts were inductively cultivated with mouse mononuclear macrophage RAW264.7, and they were divided into 2 groups： group A （control group） and group B （ALN-treated group）. The protein expressions of NFATcl, p-NF-KB and p-c-Jun were determined with Western blotting at the 2nd day of cultivation; the expression of NFATcl was assessed by immunofluorescent assay on the 4th day; and the osteoclast formation was examined on the 7th day of cultivation. RAW264.7 cells were inoculated on abrasive dentine disk, and divided into 2 groups and treated as mentioned above. The resorption function of osteoclast was observed on the 9th day of inoculation. Results TRAP positive multinuclear osteoclasts were observed, and resorption lacunaes formed in the abrasive dentine disks of the 2 groups. More TRAP positive multinuclear cells and resorption lacunaes in large size were found in group A than those in group B. Immunofluorescence assay showed the expression of NFATcl was higher in group A than in group B. The gene expressions of NFATcl, p-NF-KB and p-c-Jun were lower in group B than in group A （P〈0.01） as determined with Western blotting. Conclusion By down-regulating the expressions of p-NF-KB and p-c-Jun, ALN may strongly inhibit the osteoclast formation and its resorption function, thus inhibiting NFATcl expression.
出处
《解放军医学杂志》
CAS
CSCD
北大核心
2015年第10期778-781,共4页
Medical Journal of Chinese People's Liberation Army
基金
国家自然科学基金(81270965)
河北省自然科学基金(C2011401044)~~
