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作 者:熊琦[1] 张里程[1] 张立海[1] 姚琦[1] 唐佩福[1]
出 处:《中国骨伤》2013年第4期324-327,共4页China Journal of Orthopaedics and Traumatology
基 金:国家自然科学基金(编号:81000796;30973068)~~
摘 要: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.
关 键 词:骨保护素 核因子κb活化因子受体蛋白 破骨前体细胞 细胞分化
分 类 号:R394[医药卫生—医学遗传学]
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