Journal Article

Dynamic Regulation of Metabolism and Respiration by Endogenously Produced Nitric Oxide Protects against Oxidative Stress

Chen, Qiping, Hertkorn, Caryn, Selak, Mary, Daikhin, Evgueni, Yudkoff, Marc, Paxinou, Evgenia, Weisse, Marie, Sowa, Grzegorz, Sessa, William C., Souza, Jose M., Ischiropoulos, Harry

2001

Overview

One of the many biological functions of nitric oxide is the ability to protect cells from oxidative stress. To investigate the potential contribution of low steady state levels of nitric oxide generated by endothelial nitric oxide synthase (eNOS) and the mechanisms of protection against H2O 2, spontaneously transformed human ECV304 cells, which normally do not express eNOS, were stably transfected with a green fluorescent-tagged eNOS cDNA. The eNOS-transfected cells were found to be resistant to injury and delayed death following a 2-h exposure to H2O 2(50-150 µM). Inhibition of nitric oxide synthesis abolished the protective effect against H2O 2exposure. The ability of nitric oxide to protect cells depended on the presence of respiring mitochondria as ECV304+eNOS cells with diminished mitochondria respiration (ρ-)are injured to the same extent as nontransfected ECV304 cells and recovery of mitochondrial respiration restores the ability of nitric oxide to protect against H2O 2-induced death. Nitric oxide also found to have a profound effect in cell metabolism, because ECV304+eNOS cells had lower steady state levels of ATP and higher utilization of glucose via the glycolytic pathway than ECV304 cells. However, the protective effect of nitric oxide against H2O 2exposure is not reproduced in ECV304 cells after treatment with azide and oligomycin suggesting that the dynamic regulation of respiration by nitric oxide represent a critical and unrecognized primary line of defense against oxidative stress.

• Publisher: National Academy of Sciences,National Acad Sciences,The National Academy of Sciences
• Subject: Adenosine Triphosphate - metabolism / Azides - pharmacology / Biological Sciences / Cell Line / Cell lines / Cell Survival - drug effects / Cells / Cells, Cultured / Delta cells / Experiments / Fluorescence / Glucosephosphate Dehydrogenase - metabolism / Humans / Hydrogen peroxide / Hydrogen Peroxide - pharmacology / Kinetics / Luminescent Measurements / Mitochondria / NADP - metabolism / NG-Nitroarginine Methyl Ester - pharmacology / Nitric oxide / Nitrogen / Oligomycins - pharmacology / Oxidative Stress / Oxides / Oxygen / Oxygen consumption / Oxygen Consumption - drug effects / Physiological regulation / Respiration / Somatic cells