On the regulation of survival and death of synoviocytes and chondrocytes by nitric oxide

一氧化氮对滑膜细胞和软骨细胞生存和死亡的调节

基本信息

批准号：
16591865
负责人：
MIYAMOTO Yoichi
金额：
$ 2.18万
依托单位：
Showa University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2004
资助国家：
日本
起止时间：
2004 至 2005
项目状态：
已结题

项目摘要

It is known that nitric oxide production is up-regulated both in synovial tissues and articular cartilages in the joints of rheumatoid arthritis (RA) and osteoarthritis (OA) patients. NO is regarded as one of the important molecules to regulate cell death and survival, whereas the regulatory mechanism has not been fully elucidated. On the other hand, interleukin-1 (IL-1) acts as a key mediator of the degeneration of articular cartilage in RA and OA, where chondrocyte death is observed. In this study, the viability of mouse chondrocyte-like ATDC5 cells was reduced by the treatment with IL-1β for 48 h or longer. IL-1β augmented the expression of the catalytic subunit of NADPH-oxidase gp91^<phox> as well as inducible NO synthase in ATDC5 cells. Generation of nitrated guanosine and tyrosine suggested the formation of reactive nitrogen species including peroxynitrite (ONOO^-), a reaction product of NO and superoxide in ATDC5 cells and rat primary chondrocytes treated with IL-1β. Death of AT … More DC5 cells after IL-1β treatment was prevented by an NADPH-oxidase inhibitor 4-(2-aminoethyl) benzenesulfonyl fluoride (AEBSF), a NO synthase inhibitor N^G-nitro-L-arginine methyl ester (L-NAME), and a ONOO^- scavenger uric acid. The viability of ATDC5 cells was reduced by a ONOO^- generator 3-(4-morpholinyl)sydnonimine hydrochloride but not by either a NO donor 1-hydroxy-2-oxo-3-(N-methyl-2-aminopropyl)-3-methyl-1-triazene or S-nitrosoglutathione. Disruption of mitochondrial membrane potential and ATP deprivation were observed in IL-1β-treated ATDC5 cells, both of which were restored by L-NAME, AEBSF, or uric acid. On the other hand, any morphological or biochemical sign indicating apoptosis was not observed in these cells. These results suggest that the death of chondrocyte-like ATDC5 cells was mediated at least in part by mitochondrial dysfunction and energy depletion through ONOO^- formation after IL-1β treatment. We have also succeeded in the establishment of a fibroblastic cell line from the synoviocytes obtained from an RA patient. Less

众所周知，在类风湿关节炎（RA）和骨关节炎（OA）患者关节中，一氧化氮的产生在滑膜组织和关节软骨中都被上调。 NO被认为是调节细胞死亡和存活的重要分子之一，而调节机制尚未完全阐明。另一方面，白介素-1（IL-1）充当了RA和OA中关节软骨变性的关键介体，观察到软骨细胞死亡。在这项研究中，通过用IL-1β处理48小时或更长时间，可以降低小鼠软骨细胞样ATDC5细胞的生存能力。 IL-1β增强了NADPH-氧化酶GP91^<phox>的催化亚基以及ATDC5细胞中诱导的NO合酶的表达。硝化的鸟嘌呤和酪氨酸的产生表明形成了包括过氧亚硝酸盐（ONOO^ - ）的反应性氮，这是ATDC5细胞中NO和超氧化物的反应产物，以及用IL-1β处理的大鼠原发性软骨细胞。 NADPH-氧化酶抑制剂4-（2-氨基乙基）苯甲磺酰氟氟化物（AEBSF）预防IL-1β处理后更多的DC5细胞死亡，这是NO合成酶N^G-NITRO-L-L-精氨酸 - 精氨酸 - 精氨酸 - 甲基酯（L-NAME（L-NAME），ONOOO^-Scavenger^-Scavenger Uric Adicer。通过3-（4-苯基）盐酸盐盐的ONOO^ - 发电机降低ATDC5细胞的生存能力，但不会通过任何一个没有供体1-供体1-羟基-2-oxo-3-（N-甲基-2-氨基氨基丙基）-3-甲基-1-甲基-1-三亚烷基或s-Nentriazene或s-Nenerrosee。在IL-1β处理的ATDC5细胞中观察到了线粒体膜电位和ATP剥夺的破坏，这两种细胞均通过L-NAME，AEBSF或尿酸恢复。另一方面，在这些细胞中未观察到任何表明细胞凋亡的形态或生化体征。这些结果表明，在IL-1β处理后，通过线粒体功能障碍和通过Onoo^形成的线粒体功能障碍和能量消耗至少部分介导了软骨细胞样ATDC5细胞的死亡。我们还成功地从RA患者获得的滑膜细胞建立了成纤维细胞系。较少的