Centro Andaluz de Biología del Desarrollo, Universidad Pablo de Olavide-Consejo Superior de Investigaciones Científicas, Sevilla 41013, Spain.
FASEB J. 2011 Aug;25(8):2669-87. doi: 10.1096/fj.10-165340. Epub 2011 May 6.
Mitochondrial encephalomyopathy, lactic acidosis, and stroke-like episodes (MELAS) is a mitochondrial disease most usually caused by point mutations in tRNA genes encoded by mtDNA. Here, we report on how this mutation affects mitochondrial function in primary fibroblast cultures established from 2 patients with MELAS who harbored the A3243G mutation. Both mitochondrial respiratory chain enzyme activities and coenzyme Q(10) (CoQ) levels were significantly decreased in MELAS fibroblasts. A similar decrease in mitochondrial membrane potential was found in intact MELAS fibroblasts. Mitochondrial dysfunction was associated with increased oxidative stress and the activation of mitochondrial permeability transition (MPT), which triggered the degradation of impaired mitochondria. Furthermore, we found defective autophagosome elimination in MELAS fibroblasts. Electron and fluorescence microscopy studies confirmed a massive degradation of mitochondria and accumulation of autophagosomes, suggesting mitophagy activation and deficient autophagic flux. Transmitochondrial cybrids harboring the A3243G mutation also showed CoQ deficiency and increased autophagy activity. All these abnormalities were partially restored by CoQ supplementation. Autophagy in MELAS fibroblasts was also abolished by treatment with antioxidants or cyclosporine, suggesting that both reactive oxygen species and MPT participate in this process. Furthermore, prevention of autophagy in MELAS fibroblasts resulted in apoptotic cell death, suggesting a protective role of autophagy in MELAS fibroblasts.
线粒体脑肌病、乳酸酸中毒和卒中样发作(MELAS)是一种线粒体疾病,通常由 mtDNA 编码的 tRNA 基因突变引起。在这里,我们报告了这种突变如何影响来自 2 名 MELAS 患者的原代成纤维细胞培养物中的线粒体功能,这些患者携带 A3243G 突变。MELAS 成纤维细胞中的线粒体呼吸链酶活性和辅酶 Q10(CoQ)水平显著降低。完整的 MELAS 成纤维细胞中也发现了类似的线粒体膜电位降低。线粒体功能障碍与氧化应激增加和线粒体通透性转换(MPT)的激活有关,这触发了受损线粒体的降解。此外,我们发现 MELAS 成纤维细胞中的自噬体消除存在缺陷。电子和荧光显微镜研究证实了大量线粒体的降解和自噬体的积累,表明自噬体激活和自噬流缺陷。携带 A3243G 突变的线粒体转染细胞也显示出 CoQ 缺乏和自噬活性增加。CoQ 补充部分恢复了所有这些异常。抗氧化剂或环孢菌素处理也消除了 MELAS 成纤维细胞中的自噬,表明活性氧和 MPT 都参与了这一过程。此外,MELAS 成纤维细胞中的自噬被阻止导致凋亡细胞死亡,表明自噬在 MELAS 成纤维细胞中具有保护作用。