重组人骨保护素与重组核因子κb活化因子受体蛋白对破骨前体细胞分化的影响

摘要

Objective:To compare the effect of recombinant OPG-Fc and recombinant RANK protein on the differentiation of osteoclast precursors. Methods: Mouse osteoblasts cell lines were incubated with osteoclast precursors cell lines RAW 264.7 for 9 days with 10-5 g/L rhRANK or rhOPG-Fc or PBS added to the coculture system. TRAP stain positive cells counting and cortical bone pit formation counting were performed in the 9th day. Results: Multinuleated TRAP stain positive cells were observed in the cocluture systems after 6 days incubation, and plenty of mature osteoclasts could be observed in the 9th day. With the addition of 10-5 g/L rhOPG-Fc or rhRANK,multinucleated giant cells and cortical bone pit formation couting decreased significantly compared with the control group,and the rhRANK group decreased more significantly than the rhOPG-Fc group. Conclusions:Both rhOPG-Fc and rhRANK can inhabit the differentiation of osteoclast precursors and prevent them forming mature osteoclasts, moreover, the rhRANK shows the significant inhabition effect than the rhOPG-Fc.%目的:对比重组人骨保护素(rhOPG-Fc)与重组核因子κb活化因子受体蛋白(rhRANK)对破骨前体细胞分化的影响.方法:采用成骨细胞与破骨前体细胞RAW264.7混合培养,在地塞米松、1,25 (OH) 2VitD3诱导下生成破骨细胞的方法.研究分3组:rhRANK组:10-5 g/L；rhOPG-Fc组:10-5 g/L;空白对照组.作用9d后观察破骨细胞数目和形态,抗酒石酸酸性磷酸酶(TRAP)染色阳性细胞个数,骨磨片吸收陷窝计数.结果:在空白对照组,小鼠成骨细胞与破骨前体细胞RAW264.7混合培养6d后,开始出现多核细胞,9d时可见大量成熟多核细胞,经TRAP染色证实为成熟破骨细胞,而rhRANK组及rhOPG-Fc组TRAP染色阳性多核细胞数较对照组均减少,特别是rhRANK组减少更明显.骨片吸收陷窝计数显示rhRANK组及rhOPG-Fc组较对照组也明显减少,而相对来说,rhRANK组较rhOPG-Fc组更少.结论:rhOPG-Fc与rhRANK均可以有效抑制破骨前体细胞分化成为成熟破骨细胞,且rhRANK较rhOPG-Fc抑制效果更明显.