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

Effects of recombinant human osteoprotegerin and recombinant RANK protein on the differentiation of osteoclast precursors

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:10-5g/L;rhOPG-Fc组:10-5g/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。

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 cocuhure system. TRAP stain positive cells counting and cortical bone pit formation counting were performed in the 9th day. Results： Muhinuleated 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-s 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 rbOPG-Fc.