Mitochondrial mutagenesis correlates with the local inflammatory environment in arthritis
Mitochondrial mutagenesis correlates with the local inflammatory environment in arthritis
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DOI：
10.1136/annrheumdis-2011-200245
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发表时间：
2012-04-01
影响因子：
27.4
通讯作者：
Fearon, Ursula
中科院分区：
文献类型：
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作者：
Harty, Leonard C.;Biniecka, Monika;Fearon, Ursula
目的研究炎症性关节炎患者的线粒体突变与促炎微环境之间的关系。方法对50例炎症性关节炎患者行关节镜检查、滑膜组织活检、滑液检查和临床评估。还招募了TNFi治疗前/后的15名患者。从10名接受介入性关节镜检查的受试者中获得正常滑膜活检。通过视觉模拟量表测量宏观滑膜炎/血管分布。通过免疫组织学定量细胞特异性标志物CD 3（T细胞）和CD 68（巨噬细胞）。通过MSD多重测定法测量滑液中的TNF α、IL-6、IFN γ和IL-1 β。使用线粒体随机突变捕获试验（RMCA）对滑液组织线粒体突变进行定量。在类风湿性关节炎滑膜成纤维细胞（RASFCs）的原代培养物中评估TNF α对氧化应激和线粒体功能的直接影响。用RMCA和特异性细胞荧光探针定量检测线粒体突变、活性氧（ROS）、线粒体膜电位（MMP）和线粒体质量（MM）。
Background To examine the association between mitochondrial mutagenesis and the proinflammatory microenvironment in patients with inflammatory arthritis.Methods Fifty patients with inflammatory arthritis underwent arthroscopy and synovial tissue biopsies, synovial fluid and clinical assessment were obtained. Fifteen patients pre/post-TNFi therapy were also recruited. Normal synovial biopsies were obtained from 10 subjects undergoing interventional arthroscopy. Macroscopic synovitis/vascularity was measured by visual analogue scale. Cell-specific markers CD3 (T cells) and CD68 (macrophages) were quantified by immunohistology. TNF alpha, IL-6, IFN gamma and IL-1 beta were measured in synovial fluids by MSD multiplex assays. Synovial tissue mitochondrial mutagenesis was quantified using a mitochondrial random mutation capture assay (RMCA). The direct effect of TNF alpha on oxidative stress and mitochondrial function was assessed in primary cultures of rheumatoid arthritis synovial fibroblast cells (RASFCs). Mitochondrial mutagenesis, reactive oxygen species (ROS), mitochondrial membrane potential (MMP) and mitochondrial mass (MM) were quantified using the RMCA and specific cell fluorescent probes.Results A significant increase in mtDNA mutation frequency was demonstrated in inflamed synovial tissue compared with control (p