Galant Leticia Selinger, Doblado Laura, Radi Rafael, de Bem Andreza Fabro, Monsalve Maria
Department of Biochemistry, Federal University of Santa Catarina, Florianópolis 88040900, Brazil.
Instituto de Investigaciones Biomédicas Sols-Morreale (CSIC-UAM), Arturo Duperier 4, 28029 Madrid, Spain.
Antioxidants (Basel). 2024 Jul 20;13(7):873. doi: 10.3390/antiox13070873.
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.
了解培养细胞的复杂生物学过程，尤其是与代谢相关的过程，可能会受到培养条件的影响而产生偏差，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，细胞会降低其糖酵解通量，可作为营养物质供应受限的体外模型。然而，这些变化对相关生理过程（如氧化还原控制）的影响尚未得到充分记录，特别是在内皮细胞中，线粒体氧化被认为较低。我们评估了暴露于半乳糖或葡萄糖培养基中的内皮细胞线粒体动力学和功能的差异。我们观察到，维持在含半乳糖培养基中的细胞表现出更高的线粒体氧化能力、更融合的线粒体网络和更高的细胞间偶联。这些因素被证明会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因子的水平，发现半乳糖培养基中的牛主动脉内皮细胞（BAEC）比含葡萄糖培养基中的细胞具有更高水平的FOXO3和更低水平的Nrf2。因此，与含葡萄糖培养基相比，在内皮细胞中使用含半乳糖的培养基进行培养可能为体外线粒体相关过程的研究提供更合适的靶点；培养基会深刻影响这些细胞中的氧化还原信号传导。
