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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

已知在类风湿性关节炎（RA）和骨关节炎（OA）患者的关节中，一氧化氮的产生在滑膜组织和关节软骨中均被上调。NO被认为是调节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）在RA和OA中作为关节软骨退化的关键介质，其中观察到软骨细胞死亡。在这项研究中，小鼠软骨细胞样ATDC 5细胞的活力通过用IL-1β处理48 h或更长时间而降低。IL-1β可增强ATDC 5细胞中NADPH氧化酶催化亚基gp 91 α<phox>和诱导型NO合酶的表达。硝基化鸟苷和酪氨酸的产生表明在用IL-1β处理的ATDC 5细胞和大鼠原代软骨细胞中形成了包括过氧亚硝酸盐（ONOO^-）的活性氮物种，ONOO ^-是NO和超氧化物的反应产物。丧生的动车事故 ...更多信息 NADPH氧化酶抑制剂4-（2-氨乙基）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^-清除剂尿酸可抑制IL-1β处理后的DC 5细胞。ATDC 5细胞的存活率被ONOO^-产生剂3-（4-吗啉基）悉尼酮亚胺盐酸盐降低，但不被NO供体1-羟基-2-氧代-3-（N-甲基-2-氨丙基）-3-甲基-1-三氮烯或S-亚硝基谷胱甘肽降低。在IL-1β处理的ATDC 5细胞中观察到线粒体膜电位的破坏和ATP剥夺，这两种情况都被L-NAME、AEBSF或尿酸恢复。另一方面，在这些细胞中未观察到任何指示凋亡的形态学或生化标志。这些结果表明，软骨细胞样ATDC 5细胞的死亡至少部分由线粒体功能障碍和能量消耗介导，所述线粒体功能障碍和能量消耗通过IL-1β处理后的ONOO-β-形成。我们还成功地建立了一个成纤维细胞系从滑膜细胞从RA患者。少