摘要
目的:验证肿瘤坏死因子-α(TNF-α)是否可以直接诱外周血单核细胞(PBMCs)分化为具有骨吸收作用的破骨细胞.方法:小白鼠PBMCs体外培养中加入TNF-α和白细胞介素-1α(IL-1α)及巨噬细胞克隆集落刺激因子(M-CSF);同时,分别加入细胞核因子κB受体活化因子配体(RANKL)的拮抗剂RANK:Fc和抗TNF-α中和抗体.对培养终末细胞作组织化学染色,检测破骨细胞特征性标志物抗酒石酸磷酸酶(TRAP)的表达;并以象牙磨片上虫蚀样骨吸收陷窝的形成为指标检测其生物学活性.结果:PBMCs体外培养形成大量TRAP阳性的多核细胞(MNCs);象牙磨片上见到虫蚀样骨吸收陷窝,后者的面积对TNF-α呈剂量依赖性.RANK:Fc对此现象无抑制作用,而抗TNF-α中和抗体可完全阻断该现象的发生.结论:TNF-α能够直接诱导PBMCs分化为具有骨吸收功能的破骨细胞.
AIM： To investigate whether tumor necrosis factor-α （TNF-α） can induce osteoclast formation from peripheral blood mononuclear cells （PBMCs） independent of receptor activator of nuclear factor kappa B ligand （RANKL）. METHODS： PBMCs were cultured in the presence of macrophage-colony stimulating factor （M-CSF） and TNF-α in vitro. RANKL blocking agent RANK： Fc and neutralizing anti-TNF-α antibody were respectively added to the culture. At the end of the culture, PBMCs cultured on coverslips were fixed and stained for tartrate resistant acid phosphatase （TRAP）, a marker of osteoclast. Lacuna resorption on dentine slices, a functional marker of osteoclast, was also observed using scanning electron microscopy and light microscopy. RESULTS： TNF-α induced the formation of multinucleated cells （MNCs） which were TRAP positive and showed evidence of lacuna resorption pit formation on dentine slices. RANK： Fc did not inhibit this process while neutralizing anti-TNF-α antibody completely blocked both the formation of MNCs and lacuna resorption. CONCLUSION： In the presence of M-CSF, TNF-α can directly induce mononuclear phagocyte osteoclast precursors to differentiate into osteoclastic cells capable of lacuna resorption.
出处
《第四军医大学学报》
北大核心
2005年第21期1988-1991,共4页
Journal of the Fourth Military Medical University
