Analysis of signal transduction of osteoclast differentiation factor (RANKL) in alveolar bone destruction

破骨细胞分化因子（RANKL）在牙槽骨破坏中的信号转导分析

• 批准号: 13470394
• 负责人: UDAGAWA Nobuyuki
• 金额: $ 9.28万
• 依托单位: Matsumoto Dental University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-13470394/
• 关键词: Osteoclasts; Lipopolysaccharide; Lipopeptide; IL-1; MyD88; RANKL; OPG; Toll-like receptor; マクロファージ; リポ多糖; OPG; TNF; IL-1

Lipopolysaccharide (LPS), the major component of the outer membrane of Gram-negative bacteria, is proposed to be a potent stimulator of bone resorption in inflammatory diseases caused by bacreria. Bacterial lipoprotein/lipopeptides are also pathogen-specific molecular patterns. Recently, toll-like receptor 4 (TLR4) was identified as the signaling receptor for LPS. In addition, TLR6 associate with TLR2, and the complex of TLR6 and TLR2 recognizes diacylated mycoplasmal lipopeptides. The signaling cascade of TLR is believed to be similar to that of IL-1 receptors (IL- 1R), because both TLR and IL-1R use myeloid differentiation factor 88 (MyD88) as a common cytoplasmic signaling molecule. However, accumulating evidence also demonstrates the existence of MyD88-independent pathways, which may explain unique biological responses of individual TLR and IL-1R. Using MyD88-deficient (-/-) mice, we explored the involvement of MyD88-mediated signals in osteoclast formation. LPS, synthetic lipopept … More ide (FSL-1), IL-1a and 1.25(OH)_2D_3 all stimulated osteoclast formation in co-cultures of primary osteoblasts and bone marrow cells obtained from wild-type mice. Osteoprotegerin, a decoy receptor of RANKL, completely inhibited the osteoclast formation in the co-culture. In contrasts, LPS, lipopeptide and IL-1a failed to induce the osteoclast formation in the co-culture of Myd88 (-/-) mice-derived osteoblasts and bone marrow cells, though 1.25(OH)_2D_3 stimulated osteoclast formation even in the MyD88 (-/-) co-culture. RT-PCR analysis showed that primary osteoblasts obtained from both wild type and MyD88 (-/-) mice similarly expressed TLR2, TLR4, TLR6 and IL-1R mRNAs. LPS, lipopeptide and IL-1a stimulated expression of RANKL mRNA within 24 hr in primary osteoblasts obtained from wild-type mice but not in those from MyD88 (-/-) mice. Similarly, LPS and IL-1a stimulated phosphorylation of ERK in wild-type osteoblasts but not MyD88 (-/-) osteoblasts. Hemopoietic cells obtained from MyD88 (-/-) mice and those from wild-type mice similarly differentiated into osteoclasts within 3 days in response to RANKL and M-CSF. These results suggest that the MyD88-mediated signaling pathway is essentially involved in osteoclast formation induced by LPS, lipopeptide and IL-1a through the RANKL expression by osteoblasts. Less

脂多糖（LPS）是革兰氏阴性细菌外膜的主要成分，被认为是由巴克拉里亚引起的炎症性疾病中骨骼分辨率的潜在刺激剂。细菌脂蛋白/脂肪肽也是病原体特异性分子模式。最近，Toll样受体4（TLR4）被鉴定为LPS的信号受体。另外，TLR6与TLR2相关，TLR6和TLR2的复合物识别二核叶片脂肽。 TLR的信号传导级联反应与IL-1受体（IL-1R）相似，因为TLR和IL-1R都使用髓样分化因子88（MYD88）作为常见的细胞质信号分子。然而，积累的证据也证明了MyD88独立途径的存在，这可以解释单个TLR和IL-1R的独特生物学反应。使用MyD88缺乏（ - / - ）小鼠，我们探索了MyD88介导的信号参与破骨细胞形成。 LPS，合成脂肪……更多的IDE（FSL-1），IL-1A和1.25（OH）_2D_3在原代成骨细胞和从野生型小鼠中获得的原代成骨细胞和骨髓细胞的共培养中，所有刺激的破骨细胞形成。 RANKL的诱饵受体骨蛋白蛋白蛋白蛋白蛋白蛋白蛋白蛋白蛋白完全抑制了共培养中的破骨细胞形成。相反，LPS，Lipopeptide和IL-1A也未能在MyD88（ - / - ）小鼠衍生的破骨细胞的共培养中诱导破骨细胞的形成，即使在MyD88（ - / - ）共培养中也是如此。 RT-PCR分析表明，从野生型和MyD88（ - / - ）小鼠获得的原发性破骨细胞类似表达的TLR2，TLR4，TLR6和IL-1R mRNA。 LPS，脂蛋白肽和IL-1A在从野生型小鼠中获得的原代成骨细胞中24小时内刺激了RANKL mRNA的表达，但在MyD88（ - / - ）小鼠中刺激了RANKL mRNA。同样，LPS和IL-1A刺激了野生型成骨细胞中ERK的磷酸化，而不是MyD88（ - / - ）成骨细胞。从MyD88（ - / - ）小鼠获得的血压细胞，野生型小鼠的小鼠在3天内类似地分化为破骨细胞，以响应RANKL和M-CSF。这些结果表明，MyD88介导的信号通路基本上参与了由成骨细胞通过RANKL表达引起的LPS，脂肽和IL-1A诱导的破骨细胞形成。较少的

项目成果

Katagiri T, Takahashi N:, , 2002: "Regulatory mechanisms of osteoblast and osteoclast differentiation"Oral Diseases. (in press). (2002)

Katagiri T, Takahashi N:, , 2002：“成骨细胞和破骨细胞分化的调节机制”口腔疾病。

Li X et al.: "p38 MAPK is crucially involved in osteoclast differentiation but not in cytokine production, phagocytosis or dendritic cell differentiation of bone marrow macrophages"Endocrinology. (in press). (2003)

Li X等人：“p38 MAPK在破骨细胞分化中至关重要，但不参与骨髓巨噬细胞的细胞因子产生、吞噬作用或树突状细胞分化”内分泌学。

Takahashi N et al.: "Principles of Bone Biology, Second Edition, Volume 1"Academic press. 882 (2002)

高桥N等：《骨生物学原理，第二版，第1卷》学术出版社。

Itoh K, Udagawa N, Katagiri T, Iemura S, Ueno N, Yasuda H, Higashio K, Quinn JMW, Gillespie MT, Martin TJ, Suda T, Takahashi N.: "Bone morphogenetic protein 2 stimulates osteoclast differentiation and survival supported by receptor activator of nuclear fa

Itoh K、Udakawa N、Katagiri T、Iemura S、Ueno N、Yasuda H、Higashio K、Quinn JMW、Gillespie MT、Martin TJ、Suda T、Takahashi N.：“骨形态发生蛋白 2 通过受体支持刺激破骨细胞分化和存活

Itoh K et al.: "Lipopolysaccharide promotes the survival of osteoclasts via toll-like receptor 4, but cytokine production of osteoclasts in response to lipopolysaccharide is different from that of macrophages."J Immunol. (in press). (2003)

Itoh K 等人：“脂多糖通过 Toll 样受体 4 促进破骨细胞的存活，但破骨细胞响应脂多糖产生的细胞因子与巨噬细胞不同。”J 免疫学杂志。