Analysis of signal transduction of inflammatory cytokines in bone destruction

骨破坏中炎症细胞因子的信号转导分析

• 批准号: 12470393
• 负责人: TAKAHASHI Naoyuki
• 金额: $ 10.56万
• 依托单位: MATSUMOTO DENTAL UNIVERSITY (2001)Showa University (2000)
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2001
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-12470393/
• 关键词: Osteoclasts; Macrophage; Osteoblast; Inflammatory cytokine; p38 MAP kinase; Lipopolysaccharide; ERK; RANKL

The mechanism of osteoclastogenesis induced by lipopolysaccharide (LPS) was studied in cocultures of mouse osteoblasts and bone marrow cells. LPS stimulated osteoclastogenesis and prostaglandin E_2 (PGE_2) production in the cocultures, both of which were inhibited by NS398, a cyclooxygenase-2 inhibitor. LPS stimulated receptor activator of NF-κB ligand (RANKL) mRNA expression, and inhibited osteoprotegerin (OPG) mRNA expression in osteoblasts. NS398 inhibited only LPS-induced down-regulation of OPG mRNA expression, suggesting that LPS-stimulated PGE_2 production is important for the down-regulation of OPG. Indeed, NS398 failed to inhibit LPS-induced osteoclastogenesis in cocultures containing OPG knockout mouse-derived osteoblasts. Calcium/protein kinase C (PKC) inhibitors and PD98059 [mitogen-activated protein kinase (MAPK)/extracellular signal-regulated kinase (ERK) kinase (MEK) inhibitor] suppressed RANKL mRNA expression in osteoblasts, and inhibited osteoclastogenesis in the cocultures treated with LPS. LPS induced phosphorylation of MEK and ERK in osteoblasts, and this induction was inhibited by calcium/PKC inhibitors. In addition, PD98059 and NS398 inhibited interleukin 1 (IL-1)-induced osteoclastogenesis in the cocultures, but only PD98059 suppressed RANKL mRNA expression induced by IL-1 in osteoblasts. These results suggest that LPS and IL-1 similarly promote osteoclastogenesis through two parallel events : enhancement of RANKL expression through calcium/PKC signals followed by MEK/ERK signals, and suppression of OPG production mediated by PGE_2.

在小鼠成骨细胞和骨髓细胞的共培养物中研究了脂多糖 (LPS) 诱导的破骨细胞生成的机制，LPS 刺激了共培养物中的破骨细胞生成和前列腺素 E_2 (PGE_2) 的产生，而这两者均被环氧合酶 2 抑制剂 NS398 抑制。 LPS 刺激 NF-κB 配体受体激活剂 (RANKL) mRNA 表达，并抑制NS398 成骨细胞中的骨保护素 (OPG) mRNA 表达仅抑制 LPS 诱导的 OPG mRNA 表达下调，表明 LPS 刺激的 PGE_2 产生对于 OPG 下调很重要。 事实上，NS398 未能抑制 LPS 诱导的破骨细胞生成。在含有 OPG 敲除小鼠成骨细胞的共培养物中。 [丝裂原激活蛋白激酶 (MAPK)/细胞外信号调节激酶 (ERK) 激酶 (MEK) 抑制剂] 抑制成骨细胞中 RANKL mRNA 的表达，并抑制经 LPS 处理的共培养物中诱导的成骨细胞中 MEK 和 ERK 磷酸化的破骨细胞生成。 ，并且这种诱导被钙/PKC抑制剂抑制。此外，PD98059和NS398抑制白细胞介素1。 (IL-1) 在共培养物中诱导破骨细胞生成，但只有 PD98059 抑制成骨细胞中 IL-1 诱导的 RANKL mRNA 表达。这些结果表明，LPS 和 IL-1 通过两个平行事件类似地促进破骨细胞生成：通过钙增强 RANKL 表达。 /PKC 信号随后是 MEK/ERK 信号，以及 PGE_2 介导的 OPG 产生抑制。

项目成果

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