核因子κB受体活化因子在破骨细胞培养中表达时相的研究

Time-course expression of receptor activator of nuclear factor κB in osteoclasts culture

目的本研究旨在观察核因子κB受体活化因子(receptor activator of nuclear factorκB,RANK)在破骨细胞培养中的表达时相。方法采用巨噬细胞集落刺激因子(macrophages colony-stimulating factor,M-CSF)及核因子κB受体活化因子配体(receptor activator of NF-κB ligand,RANKL)作为分化诱导因子,诱导大鼠骨髓造血干细胞分化为破骨细胞,RT-PCR检测诱导分化不同时间点(0,3,5,7 d)破骨细胞膜表面RANK mRNA的表达水平。结果 RT-PCR检测结果显示RANK mRNA表达在培养0 d为1.20±0.47,3 d为1.52±0.58,5 d为1.59±0.57,7 d为1.69±0.51。诱导分化后表达量随培养时间延长而增加,各时间点与培养前比较均有统计学差异(P<0.05)。结论在破骨细胞分化的各个时间段RANK均有表达,是体外研究不同因素影响破骨细胞分化及活化的一个特异性指标。

Objective To explore the time-course expression of receptor activator of nuclear factor κB（RANK） in osteoclast culture.Methods Using macrophages colony-stimulating factor（M-CSF） and receptor activator of NF-κB ligand（RANKL） as inducible factors,bone marrow hematopoietic stem cells from rats were induced to differentiate into the osteoclasts.RT-PCR was used to detect RANK mRNA expression in the osteoclast cell surface at different culture time points（0 d,3 d,5 d,7 d）.Results RT-PCR resucts showed RANK mRNA expression increased in a time-dependent manner,and was significantly higher at day 3-7 than that at day 0 [1.52±0.58 at day 3,1.59±0.57 at day 5 and 1.69±0.51 at day 7 vs 1.20±0.47 at day 0,P0.05].Conclusion RANK expresses at different time points of osteoclast differentiation.It is a specific indicator for in vitro studies on different influential factors of osteoclast differentiation and activation.