Chronically ischemic mouse skeletal muscle exhibits myopathy in association with mitochondrial dysfunction and oxidative damage
Chronically ischemic mouse skeletal muscle exhibits myopathy in association with mitochondrial dysfunction and oxidative damage
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DOI：
10.1152/ajpregu.90374.2008
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发表时间：
2008-07-01
影响因子：
2.8
通讯作者：
Casale, George P.
中科院分区：
文献类型：
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作者：
Pipinos, Iraklis I.;Swanson, Stanley A.;Casale, George P.
周围动脉疾病（PAD）患者中存在以线粒体病理和氧化应激为特征的肌病。PAD患者在疾病严重程度、表现方式和是否存在共病方面存在差异。在这项研究中，我们使用小鼠后肢缺血模型，分离和直接调查慢性流入动脉闭塞对骨骼肌显微解剖，线粒体功能和表达，氧化应激的影响。在C57 BL/6小鼠中通过分阶段结扎/分离股总动脉和髂动脉诱导后肢缺血，12周后收获肌肉。通过明视野显微镜检查肌肉显微解剖结构，并通过荧光显微镜测定肌肉匀浆中柠檬酸合酶活性和ATP合酶表达的线粒体含量。通过呼吸测定法分别分析电子传递链（ETC）复合物I至IV。氧化应激被评估为总蛋白羰基和4-羟基-2-壬烯醛（HNE）加合物和锰超氧化物歧化酶（MnSOD）的表达和活性的改变。与对照组相比，缺血肌肉表现出肌病的组织学特征和线粒体含量增加。ETC复合物I、III和IV在缺血肌肉中的复合物依赖性呼吸显著降低。蛋白质羰基化，HNE加合物，MnSOD表达在缺血肌肉中显着增加。MnSOD活性无明显变化，提示MnSOD失活。使用小鼠模型，我们已经证明了第一次，流入动脉闭塞单独，即。例如，在没有其他共病的情况下，引起具有线粒体功能障碍和增加的氧化应激的肌病，重现了PAD患者的肌肉病理学。
A myopathy characterized by mitochondrial pathology and oxidative stress is present in patients with peripheral arterial disease ( PAD). Patients with PAD differ in disease severity, mode of presentation, and presence of comorbid conditions. In this study, we used a mouse model of hindlimb ischemia to isolate and directly investigate the effects of chronic inflow arterial occlusion on skeletal muscle microanatomy, mitochondrial function and expression, and oxidative stress. Hindlimb ischemia was induced by staged ligation/division of the common femoral and iliac arteries in C57BL/6 mice, and muscles were harvested 12 wk later. Muscle microanatomy was examined by bright-field microscopy, and mitochondrial content was determined as citrate synthase activity in muscle homogenates and ATP synthase expression by fluorescence microscopy. Electron transport chain (ETC) complexes I through IV were analyzed individually by respirometry. Oxidative stress was assessed as total protein carbonyls and 4-hydroxy-2-nonenal (HNE) adducts and altered expression and activity of manganese superoxide dismutase (MnSOD). Ischemic muscle exhibited histological features of myopathy and increased mitochondrial content compared with control muscle. Complex-dependent respiration was significantly reduced for ETC complexes I, III, and IV in ischemic muscle. Protein carbonyls, HNE adducts, and MnSOD expression were significantly increased in ischemic muscle. MnSOD activity was not significantly changed, suggesting MnSOD inactivation. Using a mouse model, we have demonstrated for the first time that inflow arterial occlusion alone, i. e., in the absence of other comorbid conditions, causes myopathy with mitochondrial dysfunction and increased oxidative stress, recapitulating the muscle pathology of PAD patients.