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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，在类风湿性关节炎（RA）和骨关节炎（OA）患者的关节滑膜组织和关节软骨中，一氧化氮的产生都是上调的。NO被认为是调控细胞死亡和存活的重要分子之一，但其调控机制尚未完全阐明。另一方面，白细胞介素-1 （IL-1）是RA和OA中关节软骨退行性变的关键介质，其中观察到软骨细胞死亡。在本研究中，IL-1β作用48小时或更长时间后，小鼠软骨细胞样ATDC5细胞的活力降低。IL-1β增加了ATDC5细胞中nadph氧化酶催化亚基gp91^<phox>和诱导NO合成酶的表达。硝化鸟苷和酪氨酸的产生表明，在IL-1β处理的ATDC5细胞和大鼠原代软骨细胞中形成了活性氮物质，包括过氧亚硝酸盐（ONOO^-），这是NO和超氧化物的反应产物。nadph氧化酶抑制剂4-（2-氨基乙基）苯磺酰氟（AEBSF）、NO合成酶抑制剂N^ g -硝基- l -精氨酸甲酯（L-NAME）和ONOO^-清除剂尿酸可预防IL-1β处理后AT - DC5细胞的死亡。ATDC5细胞的活力被ONOO^-产生剂3-(4-morpholinyl) sydnon亚胺盐酸盐降低，但被NO供体1-羟基-2-氧-3-（n -甲基-2-氨基丙基）-3-甲基-1-三氮杂烯或s -亚硝基谷胱甘肽不降低。在il -1β处理的ATDC5细胞中观察到线粒体膜电位破坏和ATP剥夺，这两种情况都可以通过L-NAME、AEBSF或尿酸恢复。另一方面，在这些细胞中未观察到任何形态学或生化迹象表明凋亡。这些结果表明，在IL-1β治疗后，软骨细胞样ATDC5细胞的死亡至少部分是由线粒体功能障碍和ONOO^-形成的能量消耗介导的。我们还成功地从一位RA患者的滑膜细胞中建立了成纤维细胞系。少