Involvement of nitric oxide in the pathogenesis of adult respiratory syndrome (ARDS) and the regulation of ARDS by osteopontin

一氧化氮参与成人呼吸综合征（ARDS）的发病机制以及骨桥蛋白对ARDS的调节

• 批准号: 10670559
• 负责人: SATO Kazuhiko
• 金额: $ 1.47万
• 依托单位: Juntendo University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1998
• 资助国家: 日本
• 起止时间: 1998 至 1999
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-10670559/
• 关键词: ARDS; nitric oxide; osteopontin; macrophage; mouse; 肺障害

Adult respiratory distress syndrome (ARDS) is a noncardiogenic lung edema. Recent studies suggest that nitric oxide (NO) mediated by inducible NO synthase (iNOS) in activated macrophages is involved in the pathogenesis of ARDS. Osteopontin (OPN) is a phosphorylated glycoprotein produced by a variety of cells including macrophages. Recent findings that OPN inhibits iNOS expression led us to test the hypothesis that OPN may be a counter-regulator for iNOS in development of ARDS. To address this hypothesis, we examined expression of endogenous OPN and iNOS in the lung of a mouse model of ARDS (ARDS mice) and in a murine macrophage cell line, RAW 264.7 cells, by treatment of cells with lipopolysaccharide (LPS) and interferon-γ (IFN-γ). Mice were given LPS intratracheally to produce ARDS mice. Coinduction of iNOS and OPN mRNA was observed in the lung of ARDS mice. Interestingly, OPN mRNA was induced more slowly than iNOS mRNA. Stimulation of RAW 264.7 cells with LPS and IFN-γresulted in an increase of iNOS mRNA to maximum at 12 h after stimulation. In contrast, OPN mRNA was induced more slowly than iNOS mRNA, as was also seen in the lung of ARDS mice. Induced OPN mRNA is closely associated with increased iNOS mRNA because expression of both OPN and iNOS mRNA in RAW 264.7 cells was markedly suppressed by addition of the specific iNOS inhibitor or the NOS inhibitor. In addition, the No-releasing agent spermine NONOate enhanced induction of OPN mRNA, suggesting that NO generated by iNOS up-regulates the endogenous OPN in macrophages stimulated with LPS and IFN-γ. This up-regulation of endogenous OPN may represent a negative feedback system acting to reduce iNOS expression and may contribute, at least in part, to protect the lungs from LPS-mediated tissue injury.

成人呼吸窘迫综合征（ARDS）是一种非心源性肺水肿。最近的研究表明，活化巨噬细胞中诱导型一氧化氮合酶 (iNOS) 介导的一氧化氮 (NO) 参与了 ARDS 的发病机制。骨桥蛋白 (OPN) 是一种由包括巨噬细胞在内的多种细胞产生的磷酸化糖蛋白。 OPN 抑制 iNOS 表达的最新发现使我们检验了以下假设：OPN 可能是 ARDS 发展过程中 iNOS 的反调节剂。为了验证这一假设，我们通过用脂多糖 (LPS) 和干扰素-γ (IFN-γ) 处理细胞，检测了 ARDS 小鼠模型（ARDS 小鼠）肺部和小鼠巨噬细胞系 RAW 264.7 细胞中内源性 OPN 和 iNOS 的表达。小鼠气管内注射LPS以产生ARDS小鼠。在 ARDS 小鼠的肺中观察到 iNOS 和 OPN mRNA 的共同诱导。有趣的是，OPN mRNA 的诱导速度比 iNOS mRNA 慢。用 LPS 和 IFN-γ 刺激 RAW 264.7 细胞导致 iNOS mRNA 在刺激后 12 小时增加至最大。相比之下，OPN mRNA 的诱导速度比 iNOS mRNA 的诱导速度慢，这在 ARDS 小鼠的肺中也可见到。诱导的 OPN mRNA 与 iNOS mRNA 的增加密切相关，因为通过添加特异性 iNOS 抑制剂或 NOS 抑制剂，RAW 264.7 细胞中 OPN 和 iNOS mRNA 的表达均受到显着抑制。此外，非释放剂精胺 NONOate 增强了 OPN mRNA 的诱导，表明 iNOS 产生的 NO 上调了 LPS 和 IFN-γ 刺激的巨噬细胞中的内源性 OPN。这种内源性 OPN 的上调可能代表了一种负反馈系统，可减少 iNOS 表达，并可能至少部分有助于保护肺部免受 LPS 介导的组织损伤。

项目成果

Takahashi K: "The Carboxyl-Terminal Fragment of Osteopontin Suppresses Arginine-Glycine-Asparatic Acid-dependent Cell Adhesion."Biochem and Mol Biol Int. 46. 1081-1092 (1998)

Takahashi K：“骨桥蛋白的羧基末端片段抑制精氨酸-甘氨酸-天冬氨酸依赖性细胞粘附。”Biochem 和 Mol Biol Int。

Takahashi F, Takahashi K, Maeda K, Tominaga S and Fukuchi Y: "Osteopontin is induced by nitric oxide in RAW 264.7 cells"Biochem and Mol Biol Int Life. (in press). (2000)

Takahashi F、Takahashi K、Maeda K、Tominaga S 和 Fukuchi Y：“RAW 264.7 细胞中一氧化氮诱导骨桥蛋白”Biochem 和 Mol Biol Int Life。

Choi SH, Takahashi K, Eto H, Yoon SS and Tanabe KK: "CD44H alternative splicing in colon carcinomas influences metasratic potential"Surgical Forum. XLIX. 402-403 (1998)

Choi SH、Takahashi K、Eto H、Yoon SS 和 Tanabe KK：“结肠癌中的 CD44H 选择性剪接影响转移潜能”外科论坛。

Takahashi F: "Osteopontin is induced by nitric oxide in RAW264.7 cells"Int Union Biochem and Mol Biol Life. in press (2000)

Takahashi F：“RAW264.7 细胞中的一氧化氮诱导骨桥蛋白”Int Union Biochem 和 Mol Biol Life。

Takahashi K: "The Carboxyl-Terminal Fragment of Osteopontin Suppresses Arginine-Glycine-Asparatic Asic-dependent Cell Abhesion"Biochem and Moll MIol Int. 46. 1081-1092 (1998)

Takahashi K：“骨桥蛋白的羧基末端片段抑制精氨酸-甘氨酸-天冬氨酸 Asic 依赖性细胞粘连”Biochem 和 Moll MIol Int。