Tumor necrosis factor stimulates osteoclastogenesis from human bone marrow cells under hypoxic conditions
Tumor necrosis factor stimulates osteoclastogenesis from human bone marrow cells under hypoxic conditions
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DOI：
10.1016/j.yexcr.2013.11.020
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发表时间：
2014-02-15
影响因子：
3.7
通讯作者：
Goto, Shigemi
中科院分区：
文献类型：
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作者：
Nomura, Takayuki;Aoyama, Mineyoshi;Goto, Shigemi
骨稳态是通过成骨细胞骨形成和骨吸收之间的平衡来维持的。在这项研究中，我们使用人骨髓细胞（BMCs），以探讨在肿瘤坏死因子（TNF）存在下，低氧暴露对人破骨细胞（OC）形成的作用。在存在NF-κ B配体受体激活剂（RANKL）和巨噬细胞集落刺激因子（M-CSF）的情况下，将BMC暴露于3%、5%或10% O-2，产生了抗酒石酸酸性磷酸酶（TRAP）阳性多核细胞，与OC一致。在低氧条件下添加TNF产生的成熟OC的数量显着更多的核比OC在常氧条件下产生的。较长的初始低氧暴露增加OC前体细胞的数量，并促进OC前体细胞分化为多核OC。定量RT-PCR分析显示，RANKL和TNFR 1在低氧条件下的非OC细胞中的表达水平高于常氧条件下。此外，为了证实TNF诱导的信号转导的参与，我们研究了在缺氧条件下阻断抗TNFR 1和TNFR 2抗体对OC形成的影响。观察到TNFR 1抗体显著抑制OC形成。这些结果表明，低氧暴露在TNF诱导的破骨细胞从人骨髓细胞中起着重要的作用。(C)2013 Elsevier Inc. All rights reserved.
Bone homeostasis is maintained by the balance between osteoblastic bone formation and osteoclastic bone resorption. In this study, we used human bone marrow cells (BMCs) to investigate the role of hypoxic exposure on human osteoclast (OC) formation in the presence of tumor necrosis factor (TNF). Exposing the BMCs to 3%, 5%, or 10% O-2 in the presence of receptor activator of NF-kappa B ligand (RANKL) and macrophage colony-stimulating factor (M-CSF) generated tartrate-resistant acid phosphatase (TRAP)-positive multinuclear cells, consistent with OCs. The addition of TNF under hypoxic conditions generated significantly greater numbers of mature OCs with more nuclei than OCs generated under normoxic conditions. Longer initial hypoxic exposure increased the number of OC precursor cells and facilitated the differentiation of OC precursor cells into multinucleated OCs. Quantitative RT-PCR analysis revealed that RANKL and TNFR1 were expressed at higher levels in non-OC cells from BMCs under hypoxic conditions than under normoxic conditions. Furthermore, to confirm the involvement of TNF-induced signaling, we examined the effects of blocking antibodies against TNFR1 and TNFR2 on OC formation under hypoxic conditions. The TNFR1 antibody was observed to significantly suppress OC formation. These results suggest that hypoxic exposure plays an important role in TNF-induced osteoclastogenesis from human BMCs. (C) 2013 Elsevier Inc. All rights reserved.