Reactive nitrogen and oxygen species in interleukin-1-mediated DNA damage associated with osteoarthritis
Reactive nitrogen and oxygen species in interleukin-1-mediated DNA damage associated with osteoarthritis
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DOI：
10.1016/j.joca.2007.09.012
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发表时间：
2008-05-01
影响因子：
7
通讯作者：
Fermor, B.
中科院分区：
文献类型：
--
作者：
Davies, C. M.;Guilak, F.;Fermor, B.
目的：骨关节炎（OA）与活性氮和氧以及促炎细胞因子（如白细胞介素-1（IL-1））水平升高相关。一氧化氮（NO）可介导IL-1在关节软骨中的许多分解代谢作用。本研究的目的是确定OA软骨是否显示DNA损伤的证据，如果IL-1可以诱导非OA软骨中的DNA损伤，通过增加NO或superoxide.Methods：关节软骨细胞分离自猪股骨髁和包埋在1.2%藻酸盐。使用“彗星”试验，检测与IL-1、一氧化氮合酶2（NOS 2）选择性抑制剂、自由基清除剂超氧化物歧化酶（SOD）、NO供体NOC 18或组合的NO和过氧亚硝酸盐供体SIN-1孵育24 h对DNA损伤的影响。使用Griess测定法测量NO产生。结果：OA软骨的DNA损伤明显多于非OA软骨（P < 0.001）。IL-1可引起细胞DNA损伤（P < 0.01），并伴有NO生成增加（P < 0.01）。观察到嘌呤和嘧啶的氧化DNA链断裂和碱基修饰。NOS 2抑制剂和SOD均能抑制IL-1诱导的DNA损伤（P < 0.01）。结论：骨关节炎软骨细胞存在DNA损伤，IL-1可诱导非OA软骨细胞DNA损伤及活性氧和氮自由基的产生。(C)2007年国际骨关节炎研究学会。由爱思唯尔有限公司出版。保留所有权利。
Objective: Osteoarthritis (OA) is associated with increased levels of reactive nitrogen and oxygen species and pro-inflammatory cytokines, such as interleukin-1 (IL-1). Nitric oxide (NO) can mediate a number of the catabolic effects of IL-1 in articular cartilage. The aims of this study were to determine if OA cartilage shows evidence of DNA damage, and if IL-1 could induce DNA damage in non-OA cartilage by increasing NO or superoxide.Methods: Articular chondrocytes were isolated from porcine femoral condyles and embedded in 1.2% alginate. The effects of 24 h incubation with IL-1, the nitric oxide synthase 2 (NOS2)-selective inhibitor, the free radical scavenger superoxide dismutase (SOD), the NO donor NOC18, or the combined NO and peroxynitrite donor SIN-1 on DNA damage were tested, using the "comet" assay. NO production was measured using the Griess assay. The type of oxidative damage present was assessed using a modified comet assay.Results: OA cartilage had significantly more DNA damage than non-OA cartilage (P < 0.001). IL-1 caused an increase in DNA damage (P < 0.01), which was associated with increased NO production (P < 0.01). Both oxidative DNA strand breaks and base modifications of purines and pyrimidines were observed. IL-1-induced DNA damage was inhibited by an NOS2 inhibitor or by SOD (P < 0.01). Furthermore, NOC18 or SIN-1 caused DNA damage (P < 0.001).Conclusion: Our work shows chondrocytes in osteoarthritic cartilage exhibit DNA damage, and that IL-1 induces DNA damage and reactive oxygen and nitrogen species in non-OA chondrocytes in alginate. (C) 2007 Osteoarthritis Research Society International. Published by Elsevier Ltd. All rights reserved.