ORIGINAL ARTICLE In vivo measurement of superoxide in the cerebral cortex during anoxia-reoxygenation and ischemia-reperfusion

The exact time profile of superoxide generation during anoxia-reoxygenation and ischemia reperfusion was assessed in the feline cerebral cortex in vivo using a chemiluminescence technique with a probe specific for superoxide, 2-methyl-6-[p-methoxyphenyl]-3,7-dihydroimidazo[1,2-ƒ¿]pyrazin 3-one (MCLA). MCLA solution was superfused on the cortex throughout the protocol, and MCLA chemiluminescence was measured using a newly developed photon counting system. Reflectance at 398 nm was simultaneously measured to compensate for hemodynamic artifacts resulting from cerebral blood volume changes. In 19 animals, a 90-second anoxia was induced by the inhalation of 100% ni trogen followed by a 40-minute reoxygenation. The chemiluminescence decreased during the period of anoxia (p<0.01) and then exceeded the baseline level at 15 and 20 minutes after reoxygenation (p<0.05). In six animals, superoxide dismutase (SOD) was continuously superfused and anoxia reoxygenation was performed in the same manner. The chemiluminescence decreased during the period of anoxia (p<0.05) but did not exceed the baseline level during the reoxygenation period, indicating that an increase in superoxide production was the main cause of the chemiluminescence in crease. In eight animals, a 15-minute forebrain ischemia was induced by the occlusion of the bilateral common carotid arteries with systemic hypotension (systolic blood pressure less than 50mmHg) fol lowed by a 30-minute reperfusion. The chemiluminescence decreased during the period of ischemia (p<0.01) and then increased at 20 and 25 minutes after reperfusion (p<0.05). These results indicate that superoxide generation decreases during anoxia and ischemia and then increases within 20 minutes after reoxygenation or reperfusion. (Keio J Med 51 (4): 201-207, December 2002)