Mitochondrial dysregulation of osteoarthritic human articular chondrocytes analyzed by proteomics: a decrease in mitochondrial superoxide dismutase points to a redox imbalance.
Mitochondrial dysregulation of osteoarthritic human articular chondrocytes analyzed by proteomics: a decrease in mitochondrial superoxide dismutase points to a redox imbalance.
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DOI：
10.1074/mcp.m800292-mcp200
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发表时间：
2009-01
期刊：
影响因子：
--
通讯作者：
Blanco FJ
中科院分区：
文献类型：
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作者：
Ruiz-Romero C;Calamia V;Mateos J;Carreira V;Martínez-Gomariz M;Fernández M;Blanco FJ
线粒体参与许多细胞过程；线粒体功能障碍与细胞凋亡、衰老和包括骨关节炎(OA)在内的许多病理条件有关。线粒体蛋白是研究软骨细胞代谢的有吸引力的靶点，软骨细胞是成熟软骨中存在的一种独特的细胞类型，以及它在组织降解中的作用。采用基于二维DGE和MALDI-TOF/TOF质谱仪鉴定人关节软骨细胞线粒体富含蛋白质组分的蛋白质组学方法，分析了骨性关节炎软骨细胞特有的线粒体蛋白质变化。共有73种蛋白质形式被明确地识别为在骨性关节炎中显著改变；其中23种以前被描述为线粒体。对数据进行了广泛的统计和聚类分析，揭示了骨性关节炎的线粒体蛋白图谱特征。这种模式包括能量产生的改变，维持线粒体膜的完整性，以及自由基排毒。实时定量聚合酶链式反应、免疫印迹和免疫组织化学荧光分析证实，骨性关节炎软骨表层线粒体主要抗氧化性蛋白锰超氧化物歧化酶(SOD2)显著减少。作为这种抗氧化剂缺乏的可能产物，我们发现OA软骨细胞内活性氧的产生增加，并验证了线粒体肿瘤坏死因子-α受体相关蛋白1(TRAP1)的增加，这是一种被报道具有活性氧拮抗剂作用的伴侣。我们的结果描述了正常和骨关节炎软骨细胞线粒体蛋白质谱的差异，表明在骨关节炎过程中软骨细胞发生了线粒体失调，突出了氧化还原失衡是骨关节炎发病的一个关键因素。
Mitochondria are involved in many cellular processes; mitochondrial dysfunctions have been associated with apoptosis, aging, and a number of pathological conditions, including osteoarthritis (OA). Mitochondrial proteins are attractive targets for the study of metabolism of the chondrocyte, the unique cell type present in mature cartilage, and its role in tissue degradation. Using a proteomics approach based on two-dimensional DIGE and MALDI-TOF/TOF mass spectrometric identification of mitochondria- enriched protein fractions from human articular chondrocytes, we analyzed mitochondrial protein changes that are characteristic of OA chondrocytes. A total of 73 protein forms were unambiguously identified as significantly altered in OA; 23 of them have been previously described as mitochondrial. An extensive statistical and cluster analysis of the data revealed a mitochondrial protein profile characteristic for OA. This pattern includes alterations in energy production, maintenance of mitochondrial membrane integrity, and free radical detoxification. Real time PCR, Western blot, and immunohistofluorescence assays confirmed a significant decrease of the major mitochondrial antioxidant protein manganese-superoxide dismutase (SOD2) in the superficial layer of OA cartilage. As possible outputs for this antioxidant deficiency, we found an increase of intracellular reactive oxygen species generation in OA chondrocytes and also verified an OA-dependent increase in the mitochondrial tumor necrosis factor-α receptor-associated protein 1 (TRAP1), a chaperone with a reported reactive oxygen species antagonist role. Our results describe the differences between the mitochondrial protein profiles of normal and OA chondrocytes, demonstrating that mitochondrial dysregulation occurs in cartilage cells during OA and highlighting redox imbalance as a key factor in OA pathogenesis.