Mitochondrial Oxidative Phosphorylation defect in the Heart of Subjects with Coronary Artery Disease
Mitochondrial Oxidative Phosphorylation defect in the Heart of Subjects with Coronary Artery Disease
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DOI：
10.1038/s41598-019-43761-y
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发表时间：
2019-05-20
影响因子：
4.6
通讯作者：
Beyer, Andreas M.
中科院分区：
文献类型：
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作者：
Ait-Aissa, Karima;Blaszak, Scott C.;Beyer, Andreas M.
冠状动脉疾病（CAD）是全世界死亡的主要原因，通常与线粒体功能障碍有关。目前尚缺乏对 CAD 患者线粒体功能异常的详细了解。我们评估了人类非 CAD 和 CAD 心脏中的线粒体损伤、能量产生和线粒体复合体活性。使用新鲜和冷冻的人类心脏组织。使用标准技术分离细胞裂解物或线粒体。评估线粒体 DNA ((mt)DNA)、NAD + 和 ATP 水平以及线粒体氧化磷酸化能力。还在组织裂解物中评估了对线粒体代谢和功能调节至关重要的蛋白质。 PCR 分析显示，(mt)DNA 损伤增加，线粒体常见缺失频率增加，与非 CAD 患者样本相比，两者都建立了 CAD 线粒体完整性受损的标志物。与非 CAD 受试者相比，CAD 受试者的 NAD(+) 和 ATP 水平显着降低（NAD(+) 倍数变化：非 CAD 1.00 +/- 0.17 对比 CAD 0.32 +/- 0.12*，ATP 倍数变化：非 CAD 1.00 +/- 0.294 对比 CAD 0.01 +/- 0.001*；N = 15，P < 0.005）。我们观察到 CAD 组织中呼吸控制指数降低，复合物 I、II 和 III 的活性降低。 ETC复合体亚基的表达和呼吸体的形成增加；然而，在 CAD 中观察到复合物 I 的失活形式升高。我们观察到糖酵解通量相应增加，由丙酮酸激酶和乳酸脱氢酶活性的增加表明，表明细胞能量的糖酵解补偿性增加。总之，这些结果表明，在患有 CAD 的人类心脏受试者中，线粒体代谢从氧化磷酸化转变为糖酵解。
Coronary artery disease (CAD) is a leading cause of death worldwide and frequently associated with mitochondrial dysfunction. Detailed understanding of abnormalities in mitochondrial function that occur in patients with CAD is lacking. We evaluated mitochondrial damage, energy production, and mitochondrial complex activity in human non-CAD and CAD hearts. Fresh and frozen human heart tissue was used. Cell lysate or mitochondria were isolated using standard techniques. Mitochondrial DNA ((mt)DNA), NAD + and ATP levels, and mitochondrial oxidative phosphorylation capacity were evaluated. Proteins critical to the regulation of mitochondrial metabolism and function were also evaluated in tissue lysates. PCR analysis revealed an increase in (mt)DNA lesions and the frequency of mitochondrial common deletion, both established markers for impaired mitochondrial integrity in CAD compared to non-CAD patient samples. NAD(+) and ATP levels were significantly decreased in CAD subjects compared to Non-CAD (NAD(+) fold change: non-CAD 1.00 +/- 0.17 vs. CAD 0.32 +/- 0.12* and ATP fold change: non-CAD 1.00 +/- 0.294 vs. CAD 0.01 +/- 0.001*; N = 15, P < 0.005). We observed decreased respiration control index in CAD tissue and decreased activity of complexes I, II, and III. Expression of ETC complex subunits and respirasome formation were increased; however, elevations in the de-active form of complex I were observed in CAD. We observed a corresponding increase in glycolytic flux, indicated by a rise in pyruvate kinase and lactate dehydrogenase activity, indicating a compensatory increase in glycolysis for cellular energetics. Together, these results indicate a shift in mitochondrial metabolism from oxidative phosphorylation to glycolysis in human hearts subjects with CAD.