Assessing bioenergetic function in response to oxidative stress by metabolic profiling

作者：Dranka Brian P; Benavides Gloria A; Diers Anne R; Giordano Samantha; Zelickson Blake R; Reily Colin; Zou Luyun; Chatham John C; Hill Bradford G; Zhang Jianhua; Landar Aimee; Darley Usmar Victor M^*

来源：Free Radical Biology and Medicine, 2011, 51(9): 1621-1635.

DOI：10.1016/j.freeradbiomed.2011.08.005

• Extracellular flux
• Oxygen consumption rate
• Mitochondrial profile
• Reserve capacity
• Mitochondria
• Oxidative stress
• Nitric oxide
• 4-Hydroxynonenal
• Free radicals

It is now clear that mitochondria are an important target for oxidative stress in a broad range of pathologies, including cardiovascular disease, diabetes, neurodegeneration, and cancer. Methods for assessing the impact of reactive species on isolated mitochondria are well established but constrained by the need for large amounts of material to prepare intact mitochondria for polarographic measurements. With the availability of high-resolution polarography and fluorescence techniques for the measurement of oxygen concentration in solution, measurements of mitochondrial function in intact cells can be made. Recently, the development of extracellular flux methods to monitor changes in oxygen concentration and pH in cultures of adherent cells in multiple-sample wells simultaneously has greatly enhanced the ability to measure bioenergetic function in response to oxidative stress. Here we describe these methods in detail using representative cell types from renal, cardiovascular, nervous, and tumorigenic model systems while illustrating the application of three protocols to analyze the bioenergetic response of cells to oxidative stress.

• 出版日期2011-11-1