PDF(Pubmed)
Abstract：
Understanding the complex biological processes of cells in culture, particularly those related to metabolism, can be biased by culture conditions, since the choice of energy substrate impacts all of the main metabolic pathways. When glucose is replaced by galactose, cells decrease their glycolytic flux, working as an in vitro model of limited nutrient availability. However, the effect of these changes on related physiological processes such as redox control is not well documented, particularly in endothelial cells, where mitochondrial oxidation is considered to be low. We evaluated the differences in mitochondrial dynamics and function in endothelial cells exposed to galactose or glucose culture medium. We observed that cells maintained in galactose-containing medium show a higher mitochondrial oxidative capacity, a more fused mitochondrial network, and higher intercellular coupling. These factors are documented to impact the cellular response to oxidative stress. Therefore, we analyzed the levels of two main redox regulators and found that bovine aortic endothelial cells (BAEC) in galactose media had higher levels of FOXO3 and lower levels of Nrf2 than those in glucose-containing media. Thus, cultures of endothelial cells in a galactose-containing medium may provide a more suitable target for the study of in vitro mitochondrial-related processes than those in glucose-containing media; the medium deeply influences redox signaling in these cells.
摘要：
了解培养细胞的复杂生物过程，特别是那些与新陈代谢有关的，可能会受到文化条件的偏见，因为能量底物的选择会影响所有主要的代谢途径。当葡萄糖被半乳糖取代时，细胞减少糖酵解通量，作为有限的营养可用性的体外模型。然而,这些变化对相关生理过程如氧化还原控制的影响没有很好的记录，特别是在内皮细胞中，线粒体氧化被认为是低的。我们评估了暴露于半乳糖或葡萄糖培养基的内皮细胞中线粒体动力学和功能的差异。我们观察到，维持在含半乳糖的培养基中的细胞显示出更高的线粒体氧化能力，一个更融合的线粒体网络，和更高的细胞间偶联。这些因素被证明影响细胞对氧化应激的反应。因此,我们分析了两种主要氧化还原调节剂的水平，发现半乳糖培养基中的牛主动脉内皮细胞（BAEC）的FOXO3水平高于含葡萄糖培养基中的FOXO3水平，而Nrf2水平较低。因此，与含葡萄糖的培养基相比，在含半乳糖的培养基中培养内皮细胞可能为研究体外线粒体相关过程提供更合适的靶标；培养基会深刻影响这些细胞中的氧化还原信号。
