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目的研究1α,25-二羟维生素D_3(1α,25-(OH)_2D_3)对破骨细胞(osteoclast,OC)形成及骨吸收活性的影响。方法在巨噬细胞集落刺激因子(macrophage colony stimulating factor,M-CSF)和核因子-κB受体活化因子配体(receptor activator for nuclear factor-иB ligand,RANKL)诱导破骨细胞前体细胞RAW264.7细胞分化为OC的基础上,添加不同浓度1α,25-(OH)_2D_3(10~(-7),10~(-8)和10~(-9)mol/L),通过抗酒石酸酸性磷酸酶(tartrate resistant acid phosphatase,TRAP)染色鉴定成熟OC的形成,环境扫描电镜观察牛皮质骨片吸收陷窝。结果各浓度1α,25-(OH)_2D_3均可以促进RAW264.7细胞分化为OC,并增强其骨吸收活力。其中,10~(-8)mol/L的1α,25-(OH)_2D_3诱导效果最好,所形成的OC数最多,骨吸收活性最强。结论 1α,25-(OH)_2D_3能促进OC形成,增强骨吸收活性,从而调节骨代谢。
Objective To investigate the effects of 1α, 25-dihydroxyvitamin D3 (1α, 25- (OH) _2D_3) on osteoclast formation and bone resorption. Methods Osteoclast precursor cells RAW264 were induced by macrophage colony stimulating factor (M-CSF) and receptor activator of nuclear factor-κB ligand (RANKL). 7 cells were differentiated into OC, and different concentrations of 1α, 25- (OH) _2D_3 (10 ~ (-7), 10 ~ (-8) and 10 ~ (-9) mol / L) The formation of mature OC was identified by tartrate resistant acid phosphatase (TRAP) staining, and the lacunar bovine lacunae were observed by scanning electron microscopy. Results Each concentration of 1α, 25- (OH) _2D_3 could promote the differentiation of RAW264.7 cells to OC and enhance its bone resorption activity. Among them, the induction effect of 1α, 25- (OH) _2D_3 with 10 ~ (-8) mol / L was the best, with the highest number of OC and the highest bone resorption activity. Conclusion 1α, 25- (OH) _2D_3 can promote the formation of OC, enhance the bone resorption activity, and thus regulate bone metabolism.