_.echo mitochondria 1_
The relationship between, lipid peroxidation induced by ascorbate and adenosine ADP/Fe3+, and its effect on the
respiratory chain activities of beef heart submitochondrial particles has been investigated. Lipid peroxidation, measured
as thiobarbituric acid reactive substance formation, resulted in an inhibition of the NADH and succinate oxidase
activities. Examination of several partial reactions of the respiratory chain revealed inactivation primarily of those
involving endogenous ubiquinone, i.e., NADH- and succinate-ubiquinone and cytochrome c reductases. Ubiquinol-cytochrome c
reductase, measured with reduced ubiquinone as electron donor, was unaffected. The amount of NADH- or succinate-reducible
cytochrome b in the presence of cyanide was strongly decreased, but could be recovered by the addition of antimycin. There
occurred a substantial decrease of the ubiquinone content in the course of lipid peroxidation, with a linear relationship
between this decrease and the NADH and succinate oxidase activities. The results are consistent with the conclusion that
the ubiquinone pool undergoes an oxidative modification during lipid peroxidation, to a form that can no longer function
as a component of the respiratory chain. Lipid peroxidation also led to a partial inhibition of the succinate
dehydrogenase and cytochrome c oxidase activities and a minor decrease of the cytochrome c and cytochrome a contents.
Reduction of endogenous ubiquinone prevented lipid peroxidation as well as the concomitant modification of ubiquinone and
inactivation of the respiratory chain. These observations suggest that the destruction of ubiquinone through lipid
peroxidation is the primary cause of inactivation of the respiratory chain, and emphasize the antioxidant role of
ubiquinol in preventing these effects. The possible implications of these findings for regulation of the cellular turnover
of ubiquinone by the prevailing oxidative stress are discussed. During the progression of certain degenerative conditions,
including myocardial ischemia-reperfusion injury, mitochondria are a source of increased free-radical generation and
exhibit declines in respiratory function(s). It has therefore been suggested that oxidative damage to mitochondrial
components plays a critical role in the pathology of these processes. Polyunsaturated fatty acids of membrane lipids are
prime molecular targets of free-radical damage. A major product of lipid peroxidation, 4-hydroxy-2-nonenal (HNE), is
highly cytotoxic and can readily react with and damage protein. In this study, the effects of HNE on intact cardiac
mitochondria were investigated to gain insight into potential mechanisms by which free radicals mediate mitochondrial
dysfunction. Exposure of mitochondria to micromolar concentrations of HNE caused rapid declines in NADH-linked but not
succinate-linked state 3 and uncoupled respiration. The activity of complex I was unaffected by HNE under the conditions
of our experiments. Loss of respiratory activity reflected the inability of HNE-treated mitochondria to meet NADH demand
during maximum rates of O2 consumption. HNE exerted its effects on intact mitochondria by inactivating alpha-ketoglutarate
dehydrogenase. These results therefore identify a potentially important mechanism by which free radicals bring about
declines in mitochondrial respiration.
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