Signal transduction and cell-to-cell communication in the bone resorption induced by inflammation

炎症诱导的骨吸收中的信号转导和细胞间通讯

• 批准号: 14370599
• 负责人: TAKAHASHI Naoyuki
• 金额: $ 8.83万
• 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: 2002
• 资助国家: 日本
• 起止时间: 2002 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14370599/
• 关键词: Lipopolysaccharide (LPS); IL-1; osteoclast; MyD88; Muramyl dipeptide (MDP); Nod2; TRIF; PGE_2 receptor; マクロファージ; 骨芽細胞; インターロイキン1; 樹状細胞; RANKL; p38MAPK

Aim of the study :The discovery of RANKL elucidates the mechanism of osteoclast differentiation and function regulated by osteoblasts. Using OPG-deficient (OPG-/-) mice, MyD88-deficient (MyD88-/-) mice, and TRIF-deficient (TRIF-/-) mice, we examined signal transduction and cell-to-cell communication in the bone resorption induced by inflammation PGE_2 has been shown to enhance RANKL-induced differentiation of the precursor cells into osteoclasts. We examined how PGE2 directly enhance differentiation of osteoclast progenitors. The role of PGE_2 receptors in osteoclasts was examined, using an EP4 expression vector.Results :(1)LPS stimulated the survival of purified osteoclasts through TLR4 signals. (2)p38MAP kinase was essentially involved in the differentiation of bone marrow macrophages (osteoclast precursors) into osteoclasts. The p38MAP kinase pathway was all dead in mature osteoclasts. (3)Using OPG-/-mice, we showed that IL-1 as well as LPS stimulated osteoclastogenesis through two … More parallel events : direct enhancement of RANKL expression and suppression of OPG expression. (4)Using MyD88-deficient (-/-) mice and TRF-/-mice, we showed that MyD88 signals but not TRIF signals were essentially involved in RANKL expression in osteoblasts treated with IL-1 and LPS. MyD88 signals were also essential for the survival of osteoclasts supported by LPS and IL-1. (5)Destruxin, bisphosphonates, and strontium compound S12911-2 inhibit osteoclastic bone resorption. Destruxin was shown to inhibit pit-forming activity of osteoclasts without affecting their differentiation and survival. V-ATPase induced acidification beneath the ruffled borders of osteoclasts triggers the incorporation of bisphosphonates into osteoclasts. S12911-2 was shown to inhibit ruffled border formation in osteoclasts. (6)Muramyl dipeptide (MDP) is the essential structure responsible for the immunoadjuvant activity of peptidoglycan of gram-positive and -negative bacterial walls. MDP synergistically enhanced osteoclast formation induced by LPS, IL-1α and TNF-α through RANKL expression in osteoblasts. Nod2mediated signals appear to be involved in the MDP-induced RANKL expression in osteoblasts. (7) PGE_2 has been shown to enhance RANKL-induced differentiation of the precursor cells into osteoclasts. PGE2 synergistically enhanced osteoclastic differentiation of RAW264.7cells through EP2 and EP4. In contrast, mature osteoclasts did not express functional EP2 or EP4. When EP4 cDNA was transfected into mature osteoclasts, the bone-resorbing activity was markedly inhibited by PGE_2. Less

研究的目的：RANKL的发现阐明了由成骨细胞调节的破骨细胞分化和功能的机制。使用OPG缺乏（OPG - / - ）小鼠，MyD88缺乏（MyD88 - / - ）小鼠和TRIF缺乏症（TRIF - / - ）小鼠，我们检查了由炎症PGE_2引起的骨骼分辨率的信号传递和细胞向细胞通信，已显示出可增强rankl诱导的细胞分化的先进细胞的骨骼分化。我们检查了PGE2如何直接增强破骨细胞祖细胞的分化。使用EP4表达载体检查了PGE_2受体在破骨细胞中的作用。回报：（1）LPS通过TLR4信号刺激了纯化的破骨细胞的存活。 （2）p38map激酶基本上参与了骨髓巨噬细胞（破骨细胞前体）分化为破骨细胞的分化。使用OPG - / - 小鼠的p38map激酶（3），我们表明IL-1和LPS通过两个……更平行事件刺激了整骨构成：直接增强RANKL表达和OPG表达的抑制。 （4）使用MyD88缺乏（ - / - ）小鼠和TRF - / - 小鼠，我们表明MyD88信号（而不是TRIF信号基本上都参与用IL-1和LPS处理的成骨细胞中的RANKL表达。 MYD88信号对于LPS和IL-1支持的破骨细胞的存活也至关重要。 （5）破坏蛋白，双膦酸盐和锶化合物S12911-2抑制整骨骨骨分辨率。显示毁灭素可以抑制破骨细胞的坑形成活性，而不会影响其分化和生存。 V-ATPase在破骨细胞的褶皱边界下诱导的酸化会触发双膦酸盐对破骨细胞的影响。 S12911-2被证明可以抑制破骨细胞中的褶皱边界形成。 （6）Muramyl二肽（MDP）是负责革兰氏阳性和阴性细菌壁的肽聚糖免疫辅助活性的基本结构。 MDP通过成骨细胞中的RANKL表达协同增强了由LPS，IL-1α和TNF-α诱导的破骨细胞形成。 NOD2介导的信号似乎参与了MDP诱导的破骨细胞中的RANKL表达。 （7）PGE_2已显示可增强RANKL诱导的前体细胞分化为破骨细胞。 PGE2通过EP2和EP4协同增强了RAW264.7细胞的破骨细胞分化。相反，成熟的破骨细胞没有表达功能性EP2或EP4。当将EP4 cDNA转染到成熟的破骨细胞中时，PGE_2明显抑制了骨呈现活性。较少的

项目成果

Takami M., et al.: "Involvement of vacuolar H^+-ATPase in incorporation of risedronate into osteoclasts."Bone. 32. 341-349 (2003)

Takami M.等人：“利塞膦酸盐掺入破骨细胞中液泡H 2 -ATP酶的参与。”骨。

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。

Itoh K.et al.: "LPS promotes the survival of osteoclasts via toll-like receptor 4, but cytokine production of osteoclasts in response to LPS is different from that of macrophages."Journal of Immunology. 170・7. 3688-3695 (2003)

Itoh K.等人：“LPS通过Toll样受体4促进破骨细胞的存活，但破骨细胞响应LPS的细胞因子产生与巨噬细胞不同。”免疫学杂志170・7。 2003）

Takahashi N, Udagawa N, Tanaka S, Suda T: "Generating murine osteoclasts from bone marrow."Methods Mol Med. 80. 129-144 (2003)

Takahashi N、Udakawa N、Tanaka S、Suda T：“从骨髓中生成小鼠破骨细胞。”方法 Mol Med。

Itoh K, et al.: "LPS promotes the survival of osteoclasts via toll-like receptor 4, but cytokine production of osteoclasts in response to LPS is different from that of macrophages."Journal of Immunology. 170. 3688-3695 (2003)

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