Direct stimulation of osteoclastogenesis by MIP-1α:: evidence obtained from studies using RAW264 cell clone highly responsive to RANKL
Direct stimulation of osteoclastogenesis by MIP-1α:: evidence obtained from studies using RAW264 cell clone highly responsive to RANKL
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DOI：
10.1677/joe.0.1800193
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发表时间：
2004-01-01
影响因子：
4
通讯作者：
Iijima, T
中科院分区：
文献类型：
--
作者：
Watanabe, T;Kukita, T;Iijima, T
巨噬细胞炎性蛋白-1α(MIP-1α)是CC趋化因子的一员。我们之前已经报道了使用完整的骨髓培养系统来显示MIP-1α刺激破骨细胞样多核细胞的形成。在这里，我们使用去除基质细胞的大鼠骨髓细胞和从小鼠巨噬细胞样细胞系RAW264获得的克隆来证明MIP-1α直接作用于破骨细胞系中的细胞。我们获得了几种类型的RAW264细胞克隆，其中RAW264细胞D克隆(D克隆)对NFkappaB受体激活剂配体(RANKL)和肿瘤坏死因子-α(TNF-α)表现出极高的反应性，而另一种克隆RAW264细胞N克隆(N克隆)对RANKL和TNF-α无反应。虽然两个克隆在刺激分化前都表达受体激活剂NFkappaB(RANK)，但只有D克隆在刺激细胞分化为破骨细胞时表达组织蛋白酶K。MIP-1α刺激去除基质细胞的大鼠骨髓细胞形成单核破骨样细胞。在RANKL和TNF-α刺激下，MIP-1α也能刺激D克隆的破骨细胞样多核细胞的形成。这些发现提供了强有力的证据，表明MIP-1α直接作用于破骨细胞谱系中的细胞以刺激破骨细胞的生成。此外，用MIP-1α处理RAW264细胞D克隆可显著诱导这些细胞与原代成骨细胞的黏附，提示MIP-1α在破骨细胞前体与成骨细胞在破骨细胞形成过程中的相互作用中起着重要的调节作用。
Macrophage inflammatory protein-1alpha (MIP-1alpha) is a member of the CC chemokines. We have previously reported the use of a whole bone marrow culture system to show that MIP-1alpha stimulates the formation of osteoclast-like multinucleated cells. Here we use rat bone marrow cells deprived of stromal cells, and clones obtained from murine macrophage-like cell line RAW264 to show that MIP-1alpha acts directly on cells in osteoclast lineage. We obtained several types of RAW264 cell clones, one of these clones, designated as RAW264 cell D clone (D clone), showed an extremely high response to receptor activator of NFkappaB ligand (RANKL) and tumor necrosis factor-alpha (TNF-alpha), while the other clone, RAW264 cell N clone (N clone), demonstrated no response to RANKL or TNF-alpha. Although both clones expressed receptor activator NFkappaB (RANK) before being stimulated for differentiation, only the D clone expressed cathepsin K when cells were stimulated to differentiate to osteoclasts. MIP-1alpha stimulated the formation of mononuclear preosteoclast-like cells from rat bone marrow cells deprived of stromal cells. MIP-1alpha also stimulated formation of osteoclast-like multinucleated cells from the D clone, when these cells were stimulated with RANKL and TNF-alpha. These findings provide strong evidence to show that MIP-1alpha acts directly on cells in the osteoclast lineage to stimulate osteoclastogenesis. Furthermore, pretreatment of RAW264 cell D clone with MIP-1alpha significantly induced adhesion properties of these cells to primary osteoblasts, suggesting a crucial role for MIP-1alpha in the regulation of the interaction between osteoclast precursors and osteoblasts in osteoclastogenesis.