Evidence of in vivo Free Radical Generation by Spin Trapping with α-Phenyl N-Tert-Butyl Nitrone During Ischemia/Reperfusion in Rabbit Kidneys

J. Pincemail Laboratory of Biochemistry and Radiobiology. University of Liège, Institute of Chemistry, B6, Sart Tilman, B-4000, Liège, Belgium

J. O. Defraigne Department of Transplant Surgery, University of Liège, C.H.U., B35, Sart Tilman, B-4000, Liège, Belgium

C. Franssen Department of Anaesthesiology, University of Liège. C.H.U., B33, Sart Tilman, B-4000, Liège, Belgium

T. Defechereux Department of Transplant Surgery, University of Liège, C.H.U., B35, Sart Tilman, B-4000, Liège, Belgium

J-L. Canivet Department of Anaesthesiology, University of Liège. C.H.U., B33, Sart Tilman, B-4000, Liège, Belgium

C. Philippart Laboratory of Biochemistry and Radiobiology. University of Liège, Institute of Chemistry, B6, Sart Tilman, B-4000, Liège, Belgium

M. Meurisse Department of Transplant Surgery, University of Liège, C.H.U., B35, Sart Tilman, B-4000, Liège, Belgium

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Abstract

By using α-phenyl N-tert-butyl nitrone (PBN) as spin trap molecule and the electron paramagnetic resonance (EPR) technique, we obtained the first direct evidence of in vivo intervention of free radicals during an ischemia (50 minutes) reperfusion phenomenon in kidney of an intact rabbit.

An EPR signal (triplet of doublets) characterized by coupling constants a_N = 14.75–15 G and a^H_s = 2.5–3 G was detected in blood samples. The signal was consistent with a nitroxyl-radical adduct resulting from the spin trapping by PBN of either oxygen-or carbon-centered radicals. Control experiments indicated that the EPR signal was not due to a toxic effect of the spin trap molecule.

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Evidence of in vivo Free Radical Generation by Spin Trapping with α-Phenyl N-Tert-Butyl Nitrone During Ischemia/Reperfusion in Rabbit Kidneys

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Evidence of in vivo Free Radical Generation by Spin Trapping with α-Phenyl N-Tert-Butyl Nitrone During Ischemia/Reperfusion in Rabbit Kidneys

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