Chemiluminescent measurement of increased free radical formation after ischemia/reperfusion. Mechanisms of free radical formation in the liver.
It has been proposed that xanthine oxidase-derived superoxide mediates reperfusion injury in the liver; however, there is a little direct evidence to support this hypothesis. In this paper we describe a model system to directly and noninvasively measure oxyradical formation and hepatic injury in isolated perfused rat liver. Using this sensitive chemiluminescent technique, we clearly demonstrate the theorized burst in oxygen radical production upon reperfusion of previously ischemic liver, without perturbing the system with chemical luminescence enhancers. This increase in chemiluminescence (CL) upon reperfusion was diminished by the free radical scavengers trolox and ascorbate, as well as N-2-mercaptoproprionyl-glycine (MPG), thereby confirming the oxyradical nature of this signal. Additionally, superoxide dismutase and the xanthine oxidase inhibitor allopurinol, but not catalase, attenuated the reperfusion effect, providing the most direct evidence so far that XOD derived superoxide anion is formed during liver reperfusion. Hepatic injury (AST release) did not appear to relate to increased CL, supporting the notion that the oxyradical flux may serve as a signal for other events leading to tissue injury. Further studies using this sensitive chemiluminescent technique should aid in delineating the detailed mechanism(s) of reperfusion injury.
Nunes
F A
FA
Department of Medicine, University of Pennsylvania School of Medicine, Philadelphia, USA.
Kumar
C
C
Chance
B
B
Brass
C A
CA
D000818:Animals; D001219:Aspartate Aminotransferases / Q000378:metabolism; D016166:Free Radical Scavengers / Q000494:pharmacology; D008099:Liver / Q000098:blood supply* / Q000378:metabolism*; D008163:Luminescent Measurements; D008297:Male; D010477:Perfusion; D051381:Rats; D017207:Rats, Sprague-Dawley; D015427:Reperfusion Injury / Q000378:metabolism*; D013481:Superoxides / Q000378:metabolism*; D013997:Time Factors; D014969:Xanthine Oxidase / Q000378:metabolism
D016166:Free Radical Scavengers; D013481:Superoxides; D014969:Xanthine Oxidase; D001219:Aspartate Aminotransferases