Marques Ana P, Resende Rosa, Silva Diana F, Batista Mariana, Pereira Daniela, Wildenberg Brigite, Morais Sofia, Macedo António, Pais Cláudia, Melo Joana B, Madeira Nuno, Pereira Cláudia F
Center for Innovative Biomedicine and Biotechnology (CIBB), University of Coimbra, 3004-504 Coimbra, Portugal.
Center for Neuroscience and Cell Biology (CNC), University of Coimbra, 3004-504 Coimbra, Portugal.
Biomedicines. 2021 May 7;9(5):522. doi: 10.3390/biomedicines9050522.
This study aims to evaluate whether mitochondrial changes occur in the early stages of bipolar disorder (BD). Using fibroblasts derived from BD patients and matched controls, the levels of proteins involved in mitochondrial biogenesis and dynamics (fission and fusion) were evaluated by Western Blot analysis. Mitochondrial membrane potential (MMP) was studied using the fluorescent probe TMRE. Mitochondrial morphology was analyzed with the probe Mitotracker Green and mitophagy was evaluated by quantifying the co-localization of HSP60 (mitochondria marker) and LC3B (autophagosome marker) by immunofluorescence. Furthermore, the activity of the mitochondrial respiratory chain and the glycolytic capacity of controls and BD patients-derived cells were also studied using the Seahorse technology. BD patient-derived fibroblasts exhibit fragmented mitochondria concomitantly with changes in mitochondrial dynamics and biogenesis in comparison with controls. Moreover, a decrease in the MMP and increased mitophagy was observed in fibroblasts obtained from BD patients when compared with control cells. Impaired energetic metabolism due to inhibition of the mitochondrial electron transport chain (ETC) and subsequent ATP depletion, associated with glycolysis stimulation, was also a feature of BD fibroblasts. Overall, these results support the fact that mitochondrial disturbance is an early event implicated in BD pathophysiology that might trigger neuronal changes and modification of brain circuitry.
本研究旨在评估双相情感障碍（BD）早期阶段是否发生线粒体变化。使用来自BD患者和匹配对照的成纤维细胞，通过蛋白质免疫印迹分析评估参与线粒体生物发生和动力学（分裂和融合）的蛋白质水平。使用荧光探针TMRE研究线粒体膜电位（MMP）。用探针Mitotracker Green分析线粒体形态，并通过免疫荧光定量HSP60（线粒体标志物）和LC3B（自噬体标志物）的共定位来评估线粒体自噬。此外，还使用海马技术研究了对照细胞和BD患者来源细胞的线粒体呼吸链活性和糖酵解能力。与对照相比，BD患者来源的成纤维细胞表现出线粒体碎片化，同时线粒体动力学和生物发生发生变化。此外，与对照细胞相比，在BD患者获得的成纤维细胞中观察到MMP降低和线粒体自噬增加。BD成纤维细胞的另一个特征是由于线粒体电子传递链（ETC）受到抑制以及随后的ATP消耗导致能量代谢受损，并伴有糖酵解刺激。总体而言，这些结果支持线粒体紊乱是BD病理生理学中涉及的早期事件这一事实，该事件可能触发神经元变化和脑回路改变。
