核因子-κB受体活化因子配体诱导大鼠骨髓破骨细胞形成的实验研究

• 作 者: 杨健 谭颖徽 裘松波
• 机 构: 重庆市武警医院,第三军医大学新桥医院口腔科 400061,重庆400037
• 出 处: 重庆医学
• 发表日期: 2004年03期
• 关键词: 核因子-κB受体活化因子配体 破骨细胞 骨髓 巨噬细胞集落刺激因子

目的　探讨核因子 κB受体活化因子配体 (RANKL)对成年大鼠破骨细胞生成的影响。方法　培养依赖于巨噬细胞集落刺激因子 (M CSF)的大鼠骨髓巨噬细胞作为前体破骨细胞 ,观察在含RANKL和 /或M CSF的 1 5 %胎牛血清培养基中破骨细胞的生成情况。在盖玻片和牛股骨皮质片上培养 1、3、5和 7d后 ,利用形态学观察、TRAP染色以及骨吸收陷窝检测对破骨细胞的生成进行鉴定 ,并对生成的TRAP(+)细胞进行计数和统计分析。结果　当培养细胞在RANKL和M CSF联合作用下 ,TRAP阳性染色的单核和多核破骨细胞可在 3d内迅速形成 ,骨片吸收实验显示在第 5天时骨片上出现明显的骨吸收征象 ;RAN KL呈剂量依赖型诱导TRAP阳性细胞生成。结论　在M CSF的协同作用下 ,RANKL可明显诱导成年大鼠骨髓破骨细胞的生成。

Objective To investigate the effect of RANKL on osteoclastogenesis in adult rats. Methods Rat bone marrow macrophages (MGPs) dependent on macrophage colony stimulating factor (M CSF) were cultured as precursor osteoclasts. The osteoclasts were seeded in 15% fetal bovine serum medium containing RANKL and / or M CSF The formation of the situation. After cultured for 1, 3, 5, and 7 days on the coverslips and bovine femoral cortical slices, the formation of osteoclasts was identified by morphological observation, TRAP staining and bone resorption lacuna formation, and the generated TRAP (+) cells were subjected to Counting and statistical analysis. Results When cultured cells were combined with RANKL and M CSF, TRAP positive staining of mononuclear and multinucleated osteoclasts could be rapidly formed within 3 days. The bone absorption test showed obvious bone resorption signs on the 5th day; RAN KL induced TRAP-positive cells in a dose-dependent manner. Conclusion The synergistic effect of M CSF can induce the formation of bone marrow osteoclasts in adult rats.