Inhibition of neutrophil-mediated chondrocyte cytotoxicity by nitric oxide (NO) generated by articular chondrocytes.

关节软骨细胞产生的一氧化氮 (NO) 抑制中性粒细胞介导的软骨细胞细胞毒性。

• 批准号: 06671454
• 负责人: SAURA Ryuichi
• 金额: $ 1.34万
• 依托单位: Kobe University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1994
• 资助国家: 日本
• 起止时间: 1994 至 1995
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-06671454/
• 关键词: Rheumatoid arthritis; Cartilage degradation; Chondrocyte; Polymorphonuclear leukocyte; Hydrogen peroxide; Nitric oxide; Cytokines

Nitric oxide (NO) was identified as an active product of the oxidative cleavage of L-arginine. NO is generated by endothelial cells, macrophages and neutrophils and the higher concentration of nitrite, which is the stable endproduct of NO,is detected in the rheumatoid synovial fluid and serum. It is recently reported that NO inhibits neutrophil superoxide anion production and, in this way, associated with tissue degradation in inflammatory process. We have reported that articular chondrocytes were damaged by hydrogen peroxide generated by activated neutrophils. We have studied, in this investigation, the effect of NO on neutrophil-mediated chondrocyte cytotoxicity. Primary monolayr culture of chondrocytes was obtained from bovine articular cartilage by collagenase digestion and neutrophils were separated from healthy human doners. Chondrocyte cytotoxicity was measured by chromium-51 (^<51>Cr) release from the prelabeled chondrocytes in the presence of neutrophils. The concentration of NO was measured as nitrite concentration by Grisse reaction. Articular chondrocytes synthesized NO by the stimulation of interleukin (IL-1alpha) and TNF-alpha for 24h in a dose-dependent fashion. In the presence of 100 muM of L-arginine, 200 U/ml of IL-1alpha yielded the significant generation (P<0.01) of NO (2muM nitrite equivalent) from chondrocytes. Neutrophils increased ^<51>Cr-release from articular chondrocytes, and this chondrocyte cytotoxicity was significantly (p<0.05) reversed by addition of 100 muM of L-arginine in the neutrophil-chondrocyte co-cultures In contrast, D-arginine failed to inhibit the neutrophil mediated chondrocyte cytotoxicity. These result suggested that NO generated from articular chondrocytes have a protective effect against the oxidative stress in inflammatory cartilage.

一氧化氮（NO）是l -精氨酸氧化裂解的活性产物。NO由内皮细胞、巨噬细胞和中性粒细胞产生，在类风湿滑液和血清中检测到高浓度的亚硝酸盐，亚硝酸盐是NO的稳定终产物。最近有报道称，NO抑制中性粒细胞超氧阴离子的产生，并以这种方式与炎症过程中的组织降解有关。我们曾报道过活化的中性粒细胞产生过氧化氢损伤关节软骨细胞。在这项研究中，我们研究了NO对中性粒细胞介导的软骨细胞毒性的影响。用胶原酶消化法从牛关节软骨中获得软骨细胞单层培养物，并从健康人体内分离中性粒细胞。在中性粒细胞存在的情况下，通过预先标记的软骨细胞释放的铬-51 （^<51>Cr）来测量软骨细胞的细胞毒性。用Grisse反应测定NO浓度为亚硝酸盐浓度。关节软骨细胞通过白细胞介素（il -1 α）和tnf - α刺激24小时以剂量依赖的方式合成NO。当l -精氨酸浓度为100 μ m时，il -1 α浓度为200 μ U/ml时，软骨细胞中NO（亚硝酸盐当量2muM）的生成显著（P<0.01）。在中性粒细胞-软骨细胞共培养中加入100 μ m l-精氨酸可显著（p<0.05）逆转中性粒细胞介导的软骨细胞毒性，而d -精氨酸则不能抑制中性粒细胞介导的软骨细胞毒性。提示关节软骨细胞产生的NO对炎性软骨氧化应激具有保护作用。

项目成果

Ikuo Fujita: "Inhibiton of Neutriphil-mediated Chondrosyte Cytocoxicity by Nitric Oxide Generated by Articular ChondrocYte;A Novel Protein Mechanism of the Cartilage Degractation." ORTHOPAEDIC TRANSACTIONS. 18. 502-502 (1994)

Ikuo Fujita：“通过关节软骨细胞产生的一氧化氮抑制中性粒细胞介导的软骨细胞毒性；一种软骨退化的新蛋白质机制。”

Ikuo Fuita: "Inhbition of Newtrophil-mechiated chendvocyte cytoroxicingry Nitric oxide Gcvervred by Anicular chvrdocyte : Anovel Protedies Maharisn of the Contilage Degradation." Arthvitis and Rheurvatisn. 36. S189-S189 (1993)

Ikuo Fuita：“对新生粒细胞机械化的细胞毒性一氧化氮 Gcvervred 的抑制：Contilage 降解的新型 Protedies Maharisn”。

藤田郁夫: "多核白血球による軟骨障害に対する軟骨細胞障害に対する軟骨細胞由来のNitric Oxyolaの影響" リウマチ. 33. 639-639 (1993)

Ikuo Fujita：“软骨细胞来源的硝酸 Oxyola 对多核白细胞引起的软骨细胞损伤的影响”风湿病学 33. 639-639 (1993)。

Fujita, I., et al.: "Inhibition of neutrophil-mediated chondrocyte cytotoxicity by nitric oxide generated by articular chondrocytes ; A novel protective mechanism of the cartilage degradation." Orthop.Trans.18. 502 (1994)

Fujita, I., et al.：“通过关节软骨细胞产生的一氧化氮抑制中性粒细胞介导的软骨细胞的细胞毒性；软骨退化的一种新的保护机制。”

Ikuo Fijita: "Inhbition of Newtrophil-mechiated chendvocyte cytoroxicingry Nitric oxide Gcvervred by Anicular chvrdocyte : Anovel Protedies Maharisn of the Contilage Degradation." Orthopaedic Tvanscct : ans. 18. 502-502 (1994)

Ikuo Fijita：“对新生粒细胞机械化的细胞毒性一氧化氮 Gcvervred 的抑制：Contilage 降解的 Anovel Protedies Maharisn”。