The present study was designed to measure directly, using e.p.r. spectroscopy, oxygen-derived free radicals in (post)-ischaemic or (post)-anoxic rat hearts. Rat hearts were rapidly freeze-clamped at 77 K under normoxic, anoxic, ischaemic or reperfusion conditions. The samples were measured at three different temperatures (13, 77 and 115 K) and at several microwave power levels, and were compared with isolated rat heart mitochondria. Samples were prepared both by grinding and as tissue cuts. The two preparation techniques gave identical e.p.r. results, which excludes the occurrence of grinding artifacts. No free radical signals linked to reperfusion injury were detected. Several electron transfer centres known in the mitochondrial respiratory chain were measured. The signals previously assigned to post-ischaemic reperfusion injury were found to originate from electron transfer centres of the respiratory chain, predominantly the iron-sulphur cluster S-1 in succinate dehydrogenase. The differences in signal intensity between normoxic, ischaemic and reperfused hearts were found to result from the different redox stages of these centres under the various conditions tested. These findings do not necessarily imply that oxygen-derived free radicals are not formed in cardiac tissue during (post)-ischaemic reperfusion. The constitutive background of paramagnetism from the respiratory chain, however, seriously hampers the direct detection of comparatively low concentrations of free radicals in cardiac tissue. It is therefore expedient to focus future experiments in this field on the use of spin-trapping agents.
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Research Article|
December 15 1989
Reappraisal of the e.p.r. signals in (post)-ischaemic cardiac tissue
A M van der Kraaij;
A M van der Kraaij
*Department of Biochemistry I, Erasmus University Rotterdam, P.O. Box 1738, 3000 DR Rotterdam
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J F Koster;
J F Koster
*Department of Biochemistry I, Erasmus University Rotterdam, P.O. Box 1738, 3000 DR Rotterdam
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W R Hagen
W R Hagen
†Department of Biochemistry, Agricultural University, P.O. Box 8128, 6700 ET Wageningen, The Netherlands
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Publisher: Portland Press Ltd
Online ISSN: 1470-8728
Print ISSN: 0264-6021
© 1989 London: The Biochemical Society
1989
Biochem J (1989) 264 (3): 687–694.
Citation
A M van der Kraaij, J F Koster, W R Hagen; Reappraisal of the e.p.r. signals in (post)-ischaemic cardiac tissue. Biochem J 15 December 1989; 264 (3): 687–694. doi: https://doi.org/10.1042/bj2640687
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