Formation of 4-Hydroxy-2-Nonenal-Modified Proteins in the Rat Brain Following Transient Global Ischemia Induced by Cardiac Arrest and Resuscitation
4-Hydroxy-2-nonenal (HNE) is an unsaturated aldehyde produced under conditions of oxidative stress and membrane peroxidation in many tissues. HNE forms adducts with proteins and disrupts their functions. For example, HNE can inhibit mitochondrial respiration. Proteins that are modified by reacting with HNE can be detected using antibodies raised against HNE-modified protein motifs. In this study we investigated the formation of HNE-modified proteins in rat brains subjected to transient global ischemia. Western blot analysis demonstrated the presence of HNE-modified proteins in cortex and brainstem of rats subjected to 12 min of transient global ischemia induced by cardiac arrest and resuscitation. HNE-modified proteins were detected in whole tissue samples after 15 min, 1 h, 6 h and 24 h of reperfusion after cardiac arrest and resuscitation. When compared with the whole tissue lysate at equivalent protein concentrations, the HNE-modified protein bands in the mitochondrial fraction were in the same positions but much stronger. These results suggest that HNE is produced during transient global ischemia and forms HNE-modified protein adducts in brain mitochondria which persist for at least 24 h. If HNE formation disrupts mitochondrial function in the brain it may contribute to the neuronal death that occurs after reversible global ischemia in rat brain.
KeywordsCardiac Arrest Mitochondrial Fraction Brain Mitochondrion Cereb Blood Flow Global Cerebral Ischemia
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