An altered redox balance mediates the hypersensitivity of Cockayne syndrome primary fibroblasts to oxidative stress
An altered redox balance mediates the hypersensitivity of Cockayne syndrome primary fibroblasts to oxidative stress
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DOI：
10.1111/j.1474-9726.2012.00815.x
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发表时间：
2012-06-01
期刊：
影响因子：
7.8
通讯作者：
D'Errico, Mariarosaria
中科院分区：
文献类型：
--
作者：
Pascucci, Barbara;Lemma, Tiziana;D'Errico, Mariarosaria
科凯恩综合征 (CS) 是一种罕见的遗传性多系统疾病，其特征是神经和发育障碍以及过早衰老。科凯恩综合征细胞对氧化应激高度敏感，但所涉及的分子机制仍未解决。在这里，我们提供了第一个证据，证明来自 CS-A 和 CS-B 患者的原代成纤维细胞呈现出氧化还原平衡的改变，细胞内活性氧 (ROS) 的稳态水平增加，基础和诱导的 DNA 氧化损伤、线粒体膜电位丧失以及基础氧化磷酸化速率显着降低。 Na/K-ATP酶是氧化应激的相关靶标，它也受到CS成纤维细胞转录减少的影响，并且在与野生型基因互补后恢复正常蛋白质水平。高分辨率磁共振波谱显示，与正常细胞相比，CS-A 和 CS-B 原代成纤维细胞的代谢特征明显受到干扰，这与氧化应激增加和细胞生物能学的改变一致。受影响的过程包括氧化代谢、糖酵解、胆碱磷脂代谢和渗透调节。通过在培养基中添加抗氧化剂，可以部分挽救细胞内 ROS 含量、氧化 DNA 损伤和代谢谱的变化，这表明 CS 细胞特有的持续氧化应激在潜在的病理生理学中发挥着致病作用。氧化和能量代谢的变化为CS患者的临床特征提供了线索，并为诊断和治疗提供了有价值的新工具。
Cockayne syndrome (CS) is a rare hereditary multisystem disease characterized by neurological and development impairment, and premature aging. Cockayne syndrome cells are hypersensitive to oxidative stress, but the molecular mechanisms involved remain unresolved. Here we provide the first evidence that primary fibroblasts derived from patients with CS-A and CS-B present an altered redox balance with increased steady-state levels of intracellular reactive oxygen species (ROS) and basal and induced DNA oxidative damage, loss of the mitochondrial membrane potential, and a significant decrease in the rate of basal oxidative phosphorylation. The Na/K-ATPase, a relevant target of oxidative stress, is also affected with reduced transcription in CS fibroblasts and normal protein levels restored upon complementation with wild-type genes. High-resolution magnetic resonance spectroscopy revealed a significantly perturbed metabolic profile in CS-A and CS-B primary fibroblasts compared with normal cells in agreement with increased oxidative stress and alterations in cell bioenergetics. The affected processes include oxidative metabolism, glycolysis, choline phospholipid metabolism, and osmoregulation. The alterations in intracellular ROS content, oxidative DNA damage, and metabolic profile were partially rescued by the addition of an antioxidant in the culture medium suggesting that the continuous oxidative stress that characterizes CS cells plays a causative role in the underlying pathophysiology. The changes of oxidative and energy metabolism offer a clue for the clinical features of patients with CS and provide novel tools valuable for both diagnosis and therapy.