Use of 3,3,5,5-tetramethyl-1-pyrroline-1-oxide spin trap for the continuous flow ESR monitoring of hydroxyl radical generation in the ischemic and reperfused myocardium.
A new approach for in vivo spin trapping and quantitation of oxygen-derived free radicals has been developed using a continuous flow high speed ESR detection system. Spin adducts of OH. were detected as 1:1:1:1:1:1 sextets (aN=15.2 G, aH=16.8 G, g=2.0055) in the isolated rat heart when perfused with 3,3,5,5-tetramethyl-1-pyrroline-1-oxide (40 mM) during a 10-min control pretreatment (14 ml/min) followed by 50 min of low-flow ischemia (1 ml/min), 30 min of global ischemia and subsequent reperfusion at 14 ml/min. The ESR signals appeared within 15-20 min of low-flow ischemia and grew moderately during the remaining 30 min at a rate of 2-6 nmoles of spin adduct released per minute. Post-ischemic reperfusion was characterized by a burst of spin adduct formation at 30 s-1 min, corresponding to 51.8 nmoles of spin adduct released between 30 s and 1 min.
Culcasi
M
M
Centre de Résonance Magnétique Biologique et Médicale, URA CNRS 1186, Faculté de Médecine de la Timone, Marseille, France.
Pietri
S
S
Cozzone
P J
PJ
D000818:Animals; D003327:Coronary Disease / Q000378:metabolism*; D003497:Cyclic N-Oxides; D004578:Electron Spin Resonance Spectroscopy / Q000379:methods; D005609:Free Radicals; D006878:Hydroxides / Q000378:metabolism*; D017665:Hydroxyl Radical; D066298:In Vitro Techniques; D007700:Kinetics; D008297:Male; D015425:Myocardial Reperfusion; D009206:Myocardium / Q000378:metabolism*; D051381:Rats; D011919:Rats, Inbred Strains; D013113:Spin Labels
D003497:Cyclic N-Oxides; D005609:Free Radicals; D006878:Hydroxides; D013113:Spin Labels; C051831:3,3,5,5-tetramethyl-1-pyrroline N-oxide; D017665:Hydroxyl Radical