Glucosamine Suppresses Osteoclast Differentiation through the Modulation of Glycosylation Including O-GlcNAcylation.
Glucosamine Suppresses Osteoclast Differentiation through the Modulation of Glycosylation Including O-GlcNAcylation.
复制标题
DOI：
10.1248/bpb.b16-00877
复制
发表时间：
2017-03
影响因子：
2
通讯作者：
Tomoharu Takeuchi;A. Sugimoto;N. Imazato;M. Tamura;S. Nakatani;K. Kobata;Y. Arata
中科院分区：
文献类型：
--
作者：
Tomoharu Takeuchi;A. Sugimoto;N. Imazato;M. Tamura;S. Nakatani;K. Kobata;Y. Arata
破骨细胞代表生物体中唯一的骨吸收细胞。在这项研究中，我们研究了氨基葡萄糖（GlcN），一种用于预防关节疼痛和骨质流失的营养素，对小鼠巨噬细胞样RAW 264细胞的破骨细胞生成的影响。GlcN补充抑制破骨细胞特异性基因的上调（抗酒石酸酸性磷酸酶（TRAP）、组织蛋白酶K、基质金属肽酶9和活化T细胞的核因子c1（NFATc 1）），TRAP酶活性的核因子-κB配体（RANKL）依赖性受体激活剂上调，以及TRAP阳性多核细胞的形成比N-乙酰葡糖胺（GlcNAc）更有效，我们之前已经证明其抑制破骨细胞分化。为了阐明GlcN抑制破骨细胞生成的机制，我们进一步研究了GlcN对O-GlcNAc化的影响，通过蛋白质印迹法和其他类型的糖基化凝集素印迹法。我们发现，在添加GlcN后，细胞蛋白的O-GlcNAc化增加，而α 2，6-连接的唾液酸修饰减少。因此，细胞蛋白中的这些聚糖修饰可能有助于抑制破骨细胞生成。
Osteoclasts represent the only bone resorbing cells in an organism. In this study, we investigated the effect of glucosamine (GlcN), a nutrient used to prevent joint pain and bone loss, on the osteoclastogenesis of murine macrophage-like RAW264 cells. GlcN supplementation suppressed the upregulation of osteoclast-specific genes (tartrate-resistant acid phosphatase (TRAP), cathepsin K, matrix metallopeptidase 9, and nuclear factor of activated T cell c1 (NFATc1)), receptor activator of nuclear factor-κB ligand (RANKL)-dependent upregulation of TRAP enzyme activity, and the formation of TRAP-positive multinuclear cells more effectively than N-acetylglucosamine (GlcNAc), which we have previously shown to inhibit osteoclast differentiation. To clarify the mechanism by which GlcN suppresses osteoclastogenesis, we further investigated the effect of GlcN on O-GlcNAcylation by Western blotting and on other types of glycosylation by lectin blotting. We found that, upon addition of GlcN, the O-GlcNAcylation of cellular proteins was increased whereas α2,6-linked sialic acid modification was decreased. Therefore, these glycan modifications in cellular proteins may contribute to the suppression of osteoclastogenesis.