Abstract
Detection of free radicals by electron spin resonance (ESR) proves the involvement of reactive oxygen species (ROS) in reperfused organ injuries. Spin-traps are known to ameliorate hemodynamic parameters in an isolated postischemic heart. The effects of 5 mmol/L DMPO (5,5-dimethyl-1-pyrroline-N-oxide) or DEPMPO (5-(diethylphosphoryl)-5-methyl-1-pyrrolineN-oxide) on intracellular pH (pHin) and ATP level were evaluated by31P nuclear magnetic resonance on isolated rat liver submitted to 1 hour of warm ischemia and reperfusion. At the end of the reperfusion period, during which pHin recovered to its initial value (7.16±0.03) in all groups, the ATP recovery level (expressed in percentage of initial value) was similar in controls and DEPMPO (60%±5%,n=6 and 54%±4%,n=6, respectively), but only 37%±1% in DMPO-treated livers (n=6) (p<0.05 versus controls andp<0.05 versus DEPMPO). Oxidative phosphorylation was not affected by an addition of nitrones on isolated mitochondria extracted from livers not submitted to ischemia-reperfusion. In contrast, mitochondria extracted at the end of the ischemia-reperfusion showed an impairment in the phosphorylation parameters, particularly in the presence of DMPO. Mass spectrum of ischemic liver perchloric acid extracts evidenced probable catabolites in treated groups. The differences in the effect of the two nitrones on energetic metabolism may be explained by the production of deleterious catabolites by DMPO as compared to DEPMPO. Even though a specific radical scavenging effect could be operative in the liver, our results indicate that catabolic effects were predominant. The absence of deleterious effects of DEPMPO in contrast to DMPO on the liver energetic metabolism was evidenced, allowing the use of DEPMPO for ESR detection.
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References
Jones DP (1985) The role of oxygen concentration in oxidative stress: hypoxic and hyperoxic models. InOxidative Stress (Sies H, ed.) pp. 152–195. London: Academic.
Jaeschke H, Mitchell JR (1989) Mitochondria and xanthine oxidase both generate reactive oxygen species in isolated perfused rat liver after hypoxic injury.Biochem Biophys Res Commun 160: 140–147.
Gonzalez-Flecha B, Cutrin JC, Boveris A (1993) Time course and mechanism of oxidative stress and tissue damage in rat liver subjected toin vivo ischemia-reperfusion.J Clin Invest 91: 456–464.
Rymsa B, Wang JF, De Groot H (1991) O2 − release by activated Kupffer cells upon hypoxia-reoxygenation.Am J Physiol,261: G602–607.
Kusumoto K, Morimoto T, Minor Th, Uchino J, Isselhard W (1995) Allopurinol effects in rat liver transplantation on recovery of energy metabolism and free radical-induced damage.Eur Surg Res 27: 285–291.
Sumimoto R, Jamieson NV, Kobayashi T (1991) The need for glutathione and allopurinol in HL solution for rat liver preservation.Transplantation 52: 565–567.
Arroyo CM, Kramer JH, Dickens BF, Weglicki WB (1987) Identification of free radicals in myocardial ischemia/reperfusion by spin trapping with nitrone DMPO.FEBS Lett 221: 101–104.
Zweier JL (1988) Measurement of superoxide-derived free radicals in the reperfused heart: evidence for a free radical mechanism of reperfusion injury.J Biol Chem 263: 1353–1357.
Pietri S, Culcasi M, Cozzone PJ (1989) Real-time continuous-flow spin trapping of hydroxyl free radical in the ischemic and post-ischemic myocardium.Eur J Biochem 186: 163–173.
Togashi H, Shinzawa H, Yong H, Takahashi T, Noda H, Oikawa K, Kamada H (1994) Ascorbic acid radical, superoxide, and hydroxyl radical are detected in reperfusion injury of rat liver using electron spin resonance spectroscopy.Arch Biochem Biophys 308: 1–7.
Fréjaville C, Karoui H, Tuccio B, Le Moigne F, Culcasi M, Pietri S, Lauricella R, Tordo P (1995) 5-(Diethoxyphosphoryl)-5-methyl-1-pyrrolineN-oxide; a new efficient phosphorylated nitrone for thein vitro andin vivo spin trapping of oxygen-centered radicals.J Med Chem 38: 258–265.
Tosaki A, Haseloff RF, Hellegouarch A, Schoenheit K, Martin VV, Das DK, Blasig IE (1992) Does the antia-arrhythmic effect of DMPO originate from its oxygen radical trapping property or the structure of the molecule itself?Basic Res Cardiol 87: 536–547.
Delmas-Beauvieux MC, Gallis JL, Clerc M, Canioni P (1993)31PNMR studies of rat liver cold preservation with histidine-buffered lactobionate solution.Cryobiology 30: 551–561.
Desmoulin F, Canioni P, Crotte C, Gérolami A, Cozzone PJ (1987) Hepatic metabolism during acute ethanol administration: a phosphorous-31 nuclear magnetic resonance study on perfused rat liver after normoxic or hypoxic conditions.Hepatology 7: 315–323.
Delmas-Beauvieux MC, Gallis JL, N Rousse, Clerc M, Canioni P (1992) Phosphorous-31 nuclear magnetic resonance study of isolated rat liver during hypothermic ischemia and subsequent normothermic perfusion.J Hepatol 15: 192–201.
Dufour S, Rousse N, Canioni P, Diolez P (1996) Topdown control analysis of temperature effect on oxidative phosphorylation.Biochem J 314: 743–751.
Diolez P, Moreau F (1985) Correlations between ATP synthesis, membrane potential and oxidation rate in plant mitochondria.Biochim Biophys Acta 806: 56–63.
Gallis JL, Delmas-Beauvieux MC, Biran M, Rousse N, Durand T, Canioni P (1991) Is cellular integrity responsible for the partial NMR invisibility of ATP in ischemic rat liver.NMR Biomed 4: 279–285.
Villa P, Carugo C, Guaitani A (1992) No evidence of intracellular oxidative stress during ischemia-reperfusion damage in rat liverin vivo.Tox Lett 61: 283–290.
Currin TC, Toole JG, Thurman RG, Lemasters JJ (1990) Evidence that Carolina rinse solution protects sinusoidal endothelial cells against reperfusion injury after cold ischemic storage of rat liver.Transplantation 50: 1076–1079.
Ikeda T, Yanaga K, Kishikawa K, Kakizoe S, Shimada M, Sugimachi K (1992) Ischemic injury in liver transplantation: difference in injury sites between warm and cold ischemia in rats.Hepatology 16: 454–461.
Saris NEL, Eriksson KO (1995) Mitochondrial dysfunction in ischaemia-reperfusion.Acta Anaesthesiol Scand 39: 171–176.
Bernardi P (1996) The permeability transition pore: control points of a cyclosporin A-sensitive mitochondrial channel involved in cell death.Biochim Biophys Acta 1275: 5–9.
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Delmas-Beauvieux, M.C., Pietri, S., Culcasi, M. et al. Use of spin-traps during warm ischemia-reperfusion in rat liver: comparative effect on energetic metabolism studied using31P nuclear magnetic resonance. MAGMA 5, 45–52 (1997). https://doi.org/10.1007/BF02592265
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DOI: https://doi.org/10.1007/BF02592265