Measurement by HPLC of desferrioxamine-available iron in rabbit kidneys to assess the effect of ischaemia on the distribution of iron within the total pool.
A method for the determination of desferrioxamine-available iron in tissue fractions is described which involves incubation with desferrioxamine, extraction of desferrioxamine and its iron-bound form, ferrioxamine, and quantitation of these two forms of the drug by reversed-phase hplc analysis. Chelatable iron levels in the 1-10 microMolar region could be accurately and reproducibly measured using this technique. The desferrioxamine-available iron levels in both the cortex and medulla of rabbit kidneys were significantly elevated (up to 2-fold) after the organs had been subjected to 2 hours warm ischaemia or 24 hours cold storage at 0 degrees C in hypertonic citrate solution. There was no change in the total iron content of the tissues under these circumstances and thus a redistribution of intracellular iron to more available pools had presumably taken place as a result of ischaemia. This redistribution of iron may be an important factor in the initiation of peroxidative damage to cell membranes upon reperfusion of the organ with oxygen.
Gower
J
J
Section of Surgical Research, Clinical Research Centre, Harrow, Middlesex, U.K.
Healing
G
G
Green
C
C
D000818:Animals; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D003600:Cytosol / Q000378:metabolism; D003676:Deferoxamine / Q000302:isolation & purification / Q000378:metabolism*; D007501:Iron / Q000302:isolation & purification / Q000378:metabolism*; D007511:Ischemia / Q000378:metabolism*; D007668:Kidney / Q000098:blood supply / Q000378:metabolism*; D007672:Kidney Cortex / Q000378:metabolism; D007679:Kidney Medulla / Q000378:metabolism; D007700:Kinetics; D011485:Protein Binding; D011817:Rabbits; D012016:Reference Values
D007501:Iron; D003676:Deferoxamine