Effect of oxygen tension on bioenergetics and proteostasis in young and old myoblast precursor cells

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• 作者：M. Konigsberg ; V.I. P茅rez ; C. R铆os ; Y. Liu ; S. Lee ; Y. Shi ; H. Van Remmen
• 关键词：Myoblasts ; Mitochondrial respiration ; Protein damage ; Hypoxia ; Oxidative stress
• 刊名：Redox Biology
• 出版年：2013
• 年：2013
• 卷：1
• 期：1
• 页码：475-482
• 全文大小：1316 K

文摘

In the majority of studies using primary cultures of myoblasts, the cells are maintained at ambient oxygen tension (21% O₂), despite the fact that physiological O₂ at the tissue level in vivo is much lower (~1-5% O₂). We hypothesized that the cellular response in presence of high oxygen concentration might be particularly important in studies comparing energetic function or oxidative stress in cells isolated from young versus old animals. To test this, we asked whether oxygen tension plays a role in mitochondrial bioenergetics (oxygen consumption, glycolysis and fatty acid oxidation) or oxidative damage to proteins (protein disulfides, carbonyls and aggregates) in myoblast precursor cells (MPCs) isolated from young (3-4 m) and old (29-30 m) C57BL/6 mice. MPCs were grown under physiological (3%) or ambient (21%) O₂ for two weeks prior to exposure to an acute oxidative insult (H₂O₂). Our results show significantly higher basal mitochondrial respiration in young versus old MPCs, an increase in basal respiration in young MPCs maintained at 3% O₂ compared to cells maintained at 21% O₂, and a shift toward glycolytic metabolism in old MPCs grown at 21% O₂. H₂O₂ treatment significantly reduced respiration in old MPCs grown at 3% O₂ but did not further repress respiration at 21% O₂ in old MPCs. Oxidative damage to protein was higher in cells maintained at 21% O₂ and increased in response to H₂O₂ in old MPCs. These data underscore the importance of understanding the effect of ambient oxygen tension in cell culture studies, in particular studies measuring oxidative damage and mitochondrial function.