Iron-dependent peroxidation of rat brain: a regional study.
Iron has been shown to initiate a variety of free radical reactions in biological systems. The present study examined the in vitro susceptibility of homogenates prepared from different regions of rat brain to iron-induced peroxidation. Among the regions studied, basal thiobarbituric acid-reactive product (TBAR) formation is highest in the cerebellum and amygdala, intermediate in the cortex, hippocampus, and neostratium, and lowest in the hypothalamus, midbrain, and brainstem. In the presence of 200 microM FeCl3, there is a 20-25-fold increase in the net TBAR formation in all regions, with TBAR formation in the cerebellum and amygdala being significantly higher than in the midbrain and brainstem. Time-course and dose-response studies of iron-induced peroxidation showed that the cerebellum and amygdala are the most susceptible regions with respect to concentration of iron and duration of the incubation time, whereas the midbrain and brainstem are the least affected areas. Following low-speed (1,000 g) centrifugation of brain part homogenates, TBAR formation in the supernatant fractions is quite uniform across regions, while the pellet fractions give the same regional variations as the whole homogenates. TBAR formation in both fractions is increased 20-30-fold in the presence of 200 microM iron. Brain tissue TBAR formation induced by 200 microM iron is inhibited by the iron chelator desferrioxamine (IC50 = 600 microM), by Tris buffer pH = 8.0 (2.5 mM Tris gives 50% inhibition by trapping hydroxyl radicals), and by high concentrations of the cyclooxygenase inhibitor indomethacin (IC50 = 1.2 mM).
Subbarao
K V
KV
Department of Pharmacology, College of Medicine, University of Saskatchewan, Saskatoon, Canada.
Richardson
J S
JS
D000818:Animals; D001921:Brain / Q000187:drug effects / Q000378:metabolism*; D002614:Chelating Agents / Q000494:pharmacology; D004247:DNA / Q000378:metabolism*; D003676:Deferoxamine / Q000494:pharmacology; D005609:Free Radicals; D066298:In Vitro Techniques; D007501:Iron / Q000378:metabolism*; D015227:Lipid Peroxidation / Q000187:drug effects*; D008297:Male; D051381:Rats; D011919:Rats, Inbred Strains; D013858:Thiobarbiturates
D002614:Chelating Agents; D005609:Free Radicals; D013858:Thiobarbiturates; D004247:DNA; D007501:Iron; D003676:Deferoxamine; C029684:thiobarbituric acid