不同浓度破骨细胞分化因子和1,25-(OH)_2D_3体外诱导大鼠破骨样细胞形成的研究

Induction of osteoclast-like cells using different concentrations of receptor activator of NF-κB ligand and 1,25-(OH)_2D_3

目的研究破骨细胞分化因子和1,25-(OH)2D3共同作用对大鼠破骨样细胞体外形成的影响。方法采用大鼠脾细胞和骨细胞联合培养,实验组分别加入不同浓度的破骨细胞分化因子和1,25-(OH)2D3进行诱导,并利用抗酒石酸酸性磷酸酶(TRAP)染色、骨吸收陷窝检测等方法对破骨样细胞进行鉴定,对TRAP(+)的细胞进行计数和统计学分析。统计学方法采用单因素方差分析,组间比较用SNK检验。结果各实验组细胞均有TRAP(+)多核破骨细胞出现,并在牙本质磨片上形成吸收陷窝,破骨细胞分化因子和1,25-(OH)2D3共同作用的双因子诱导TRAP(+)多核破骨细胞形成的数量要大于使用单因子诱导,且因子浓度越大,诱导效果越好。结论破骨细胞分化因子和1,25-(OH)2D3共同作用及高浓度的因子诱导可有效诱导体外破骨样细胞的形成。

Objective To study the effects of receptor activator of NF-κB ligand （RANKL） and 1,25-（OH）2D3 on the formation of osteoclast-like cells in vitro. Methods The rat spleen cells and bone cells were incubated in the same compartment. The experimental group was incubated with different concentrations of RANKL and 1,25-（OH）2D3. The TRAP staining, bone absorption lacunae detection were employed to identify the osteoclast-like cells and the number of TRAP （＋） cells. One-way ANOVA and SNK test were used for statistic analysis. Results TRAP （＋） multi-nuclear cells, in conjunction with absorption lacunae, were observed in all groups of cells. A greater number of TRAP （＋） multinuclear cells were observed in the groups treated with the combination of RANKL and 1,25-（OH）2D3, in a concentration dependent fashion. Conclusion The effects of RANKL and 1,25-（OH）2D3, in particular, at higher con-centrations, may promote the genesis and increase the absorption of osteoclasts.