Postischaemic reperfusion injury is often attributed to the generation of oxygenated free radicals which may subsequently promote lipid peroxidation in cell membranes. Electron paramagnetic resonance spectroscopy in association with the spin trap molecule α-phenyl-N-tert-butyl-nitrone allowed direct confirmation of lipid free radical production after renal ischaemia-reperfusion in an in vivo rabbit model. A 60-min period of ischaemia followed by reperfusion caused free radical production twofold greater than after 15min of ischaemia. Glutathione and a-tocopherol have been measured in renal tissue, as indirect markers of lipid peroxidation. After 15min of ischaemia followed by 10min of reperfusion, the mean(s.e.m.) glutathione content of the ischaemic kidney was slightly but significantly reduced by 11.9(2.5)% (P < 0.003). The content of α-tocopherol was unchanged. However, 10min of reperfusion following 60 min of ischaemia led to significant decrease in mean(s.e.m.) content of both glutathione (30.4(3.7)%) (2.23(0.2) versus 3.14(0.18) μmol/g wet tissue, (P < 0.001) and α-tocopherol (46.1(7.8)%) (0.57(0.10) versus 1.09(0.14) μg/g wet tissue. P < 0.001) when compared to the control kidney. Under these experimental conditions, desferrioxamine (15mg/kg administered intravenously before inducing ischaemia), a drug known to limit free radical production, significantly limited the decrease of α-tocopherol to 20.8(6.4)% (0.83(0.08) versus 1.05(0.04) μg/g wet tissue. P < 0.05), but did not prevent glutathione consumption in the reperfused kidney.
McCordJM. Oxygen-derived free radicals in postischemic tissue injury. N Engl J Med1985; 312: 159–63.
2.
ParksDA, BulkleyGB, GrangerDN, HamiltonSR, McCordJM. Ischemic injury in the cat small intestine: role of superoxide radicals. Gastroenterology1982; 82: 9–15.
3.
AtallaSL, Toledo-PereyraIH, MackenzieGH, CerdenaJP. Influence of oxygen-derived free radical scavengers on ischemic livers. Transplantation1985; 40: 585–90.
4.
PallerMS, HerbelRP. Ethane production as a measure of lipid peroxidation after renal ischemia. Am J Physiol1986; 251: 839–43.
5.
CooperAL, PulsinelliWA, DuffyTH. Glutathione and ascorbate during ischemia and postischemic reperfusion in rat brain. J Neurochem1980; 35: 1242–5.
6.
McCoyRN, HillKE, AyonMA, SteinJH, BurhRF. Oxidant stress following renal ischemia: changes in the glutathione redox ratio. Kidney Int1988; 33: 812–17.
7.
MarubayashiS, DohiK, YamadaK, KawasakiT.Changes in the levels of endogenous coenzyme Q homologs, α-tocopherol, and glutathione in rat liver after hepatic ischemia and reperfusion, and the effect of pretreatment with coenzyme Q10. Biochim Biophys Acta1984; 797: 1–9.
8.
PuettDW, FormanMB, CatesCUOxypurinol limits myocardial stunning but does not reduce infarct size after reperfusion. Circulation1987; 76: 678–86.
9.
BandoK, TeramotoS, TagoMOxygenated perfluorocarbon, recombinant human superoxide dismutase, and catalase ameliorate free radical induced myocardial injury during heart preservation and transplantation. J Thorac Cardiovasc Surg1988; 96: 930–8.
10.
HanssonR, GustafssonB, JonssonOEffect of xanthine oxidase inhibition on renal circulation after ischemia. Transplant Proc1982; XIV: 51–8.
11.
ZweierJL, FlahertyJT, WeisfeldtML. Direct measurement of free radical generation following reperfusion of ischemic myocardium. Proc Natl Acad Sci USA1987; 84: 1404–7.
12.
GarlickPB, DaviesMJ, HearseDJ, SlaterTF. Direct detection of free radicals in the reperfused rat heart using electron spin resonance spectroscopy. Circ Res1987; 61: 757–60.
13.
BF, WeglickiWB. Identification of free radicals in myocardial ischemia/reperfusion by spin trapping with nitrone DMPO. FEBS Lett1987; 221: 101–4.
RaoPS, CohenMV, MuellerHS. Production of free radicals and lipid peroxides in early experimental myocardial ischemia. J Mol Cell Cardiol1983; 15: 713–16.
16.
ArroyoCM, KramerJH, LeiboffRH, MergnerGW, DickensBF, WeglickiWB. Spin trapping of oxygen and carbon centered free radicals in ischemic canine myocardium. Free Radic Biol Med1987; 3: 313–16.
17.
BolliR, PatelBS, JeroudiMO, LaiEK, McCayPB. Demonstration of free radicals generation in «stunned’ myocardium of intact dogs with the use of the spin trapα-phenyl N-tert-butyl nitrone. J Clin Invest1988; 82: 476–85.
18.
NilssonUA, LundgrenO, HaglindE, Bylund-FelleniusAC. Radical production during in vivo intestinal ischemia and reperfusion in cat. Am J Physiol1989; 20: 409–14.
19.
FolchJ, LeeM, Shane-StanleyGH. Simple method for the isolation and purification of total lipid animal tissues. J Biol Chem1957; 266: 497–510.
20.
SedlakJ, LindsayRH. Estimation of total protein bound and non-protein sulfhydryl groups in tissue with Ellman's reagent. Anal Biochem1968; 25: 192–205.
21.
ChouPP, JaynesPK, BaileyJL. Determination of vitamin E in microsamples of serum by liquid chromatography with electrochemical detection. Clin Chem1985; 31: 880–2.
22.
GreenGJ, HealingG, SimpkinS, LunecJ, FullerBJ. Desferrioxamine reduces susceptibility to lipid peroxidation in rabbit kidneys subjected to warm ischemia and reperfusion. Corop Biochem Physiol1986; 85B: 113–17.
23.
GreenCL, HealingG, LunecJ, FullerBJ, SimpkinS.Evidence of free-radical-induced damage in rabbit kidneys after simple hypothermic preservation and autotransplantation. Transplantation1986; 41: 161–5.
24.
HoshinoT, MaleyWR, BulkleyGB, WilliamsGM. Ablation of free radical-mediated reperfusion injury for the salvage of kidneys taken from non-heartbeating donors. Transplantation1988; 45: 428–9.
25.
KoyamaI, BulkleyGB, WilliamsGM, ImMJ. The role of oxygen free radicals in mediating the reperfusion injury of cold-preserved ischemic kidneys. Transplantation1985; 40: 590–5.
SiemsW, MielheB, MullerM, HermannC, RäderL, GerberG.Status of glutathione in the rat liver. Enhanced formation of oxygen radicals at low oxygen tension. Biomed Biochim Acta1983; 42: 1079–89.
28.
FerrariR, CecariC, CurelloSOxygen-mediated myocardial damage during ischemia and reperfusion: role of the cellular defenses against oxygen toxicity. J Mol Cell Cardiol1985; 17: 937–45.
29.
LinasSL, ShanleyPF, WhiteCW, ParkerNP, RepineJE. O2 metabolite-mediated injury in perfused kidneys is reflected by consumption of DMTU and glutathione. Am J Physiol1987; 253: 692–701.
30.
YoshidaSH, BustoR, WatsonBD, SantisoM, GinsbergMD. Postischemic cerebral lipid peroxidation in vitro: modification by dietary vitamin E. J Neurochem1985; 44: 1593–601.
31.
GowerJ, FullerB, GreenC.Lipid peroxidation in the cortex and medulla of rabbit kidneys subjected to cold ischemia and the value of protective agents. Free Radic Res Com1987; 3: 107–15.
32.
OwenML, LazarusHM, WolcottMW, MaxwellJG, TaylorJB. Allopurinol and hypoxanthine pretreatment of canine kidney donors. Transplantation1974; 17: 424–7.
33.
HalliwellB, GutteridgeJMC. The importance of free radicals and catalytic metal ions in human diseases. Mol Aspects Med1985; 8: 89–113.
34.
PallerMS, HedlundBE, SckoraJ, FaassenA, WaterfieldR.Role of iron in postischemic renal injury in the rat. Kidney Int1988; 34: 474–80.
35.
FullerBJ, LunecJ, HealingG, SimpkinS, GreenCJ. Reduction of susceptibility to lipid peroxidation by desferrioxamine in rabbit kidneys subjected to 24-hour cold ischemia and reperfusion. Transplantation1986; 43: 604–6.
