Mordhorst Bethany R, Murphy Stephanie L, Schauflinger Martin, Rojas Salazar Shirley, Ji Tieming, Behura Susanta K, Wells Kevin D, Green Jonathan A, Prather Randall S
1 Department of Animal Sciences, University of Missouri , Columbia, Missouri.
2 Electron Microscopy Core Facility, University of Missouri , Columbia, Missouri.
Cell Reprogram. 2018 Aug;20(4):225-235. doi: 10.1089/cell.2018.0008.
The Warburg effect is characterized by decreased mitochondrial oxidative phosphorylation and increased glycolytic flux in adequate oxygen. The preimplantation embryo has been described to have characteristics of the Warburg effect, including similar changes in gene expression and mitochondria, which are more rudimentary in appearance. We hypothesized hypoxia would facilitate anaerobic glycolysis in fibroblasts thereby promoting gene expression and media metabolite production reflecting the Warburg effect hallmarks in early embryos. Additionally, we speculated that hypoxia would induce a rudimentary small mitochondrial phenotype observed in several cell types evidenced to demonstrate the Warburg effect. While many have examined the role hypoxia plays in pathological conditions, few studies have investigated changes in primary cells which could be used in somatic cell nuclear transfer. We found that cells grown in 1.25% O had normal cell viability and more, but smaller mitochondria. Several hypoxia-inducible genes were identified, including seven genes for glycolytic enzymes. In conditioned media from hypoxic cells, the quantities of gluconolactone, cytosine, and uric acid were decreased indicating higher consumption than control cells. These results indicate that fibroblasts alter gene expression and mitochondria to compensate for hypoxic stress and maintain viability. Furthermore, the metabolic changes observed, making them more similar to preimplantation embryos, could be facilitating nuclear reprogramming making these cells more amendable to future use in somatic cell nuclear transfer.
瓦伯格效应的特征是在充足氧气条件下线粒体氧化磷酸化减少以及糖酵解通量增加。着床前胚胎已被描述具有瓦伯格效应的特征，包括基因表达和线粒体的类似变化，其外观更为原始。我们假设缺氧会促进成纤维细胞中的无氧糖酵解，从而促进反映早期胚胎中瓦伯格效应特征的基因表达和培养基代谢产物的产生。此外，我们推测缺氧会诱导在几种细胞类型中观察到的原始小线粒体表型，这已被证明可证明瓦伯格效应。虽然许多人研究了缺氧在病理状况中所起的作用，但很少有研究调查可用于体细胞核移植的原代细胞的变化。我们发现，在1.25%氧气环境中生长的细胞具有正常的细胞活力，并且线粒体更多但更小。鉴定出了几个缺氧诱导基因，包括七个糖酵解酶基因。在缺氧细胞的条件培养基中，葡萄糖酸内酯、胞嘧啶和尿酸的量减少，表明其消耗量高于对照细胞。这些结果表明，成纤维细胞改变基因表达和线粒体以补偿缺氧应激并维持活力。此外，观察到的代谢变化使其更类似于着床前胚胎，这可能有助于核重编程，使这些细胞更适合未来用于体细胞核移植。
