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Chemiluminescent measurement of increased free radical formation after ischemia/reperfusion

Mechanisms of free radical formation in the liver

  • Liver: Infectious, Infiammatory, And Metabolic Disorders
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Abstract

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.

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Authors and Affiliations

  1. Department of Biochemistry and Biophysics, University of Pennsylvania School of Medicine, and the Philadelphia Veterans Administration Hospital, Philadelphia, Pennsylvania

    Frederick A. Nunes MD, Chellappa Kumar PhD, Britton Chance PhD &  Clifford A. Brass MD, PhD

  2. Division of Gastroenterology, University of Pennsylvania, Clinical Research Building 638b, 422 Curie Boulevard, 19104-6144, Philadelphia, Pennsylvania

    Frederick A. Nunes MD, Chellappa Kumar PhD, Britton Chance PhD &  Clifford A. Brass MD, PhD

Authors
  1. Frederick A. Nunes MD
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  2. Chellappa Kumar PhD
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  3. Britton Chance PhD
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  4. Clifford A. Brass MD, PhD
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Additional information

C. A. Brass was the Hans Popper Liver Scholar of the American Liver Foundation (1989–1993). Supported in part by a VA Merit Review Award and NIH training grant 5-21643 (F.A. Nunes). Presented in part at the annual meeting of the American Association for the Study of Liver Disease, November, 1992.

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Nunes, F.A., Kumar, C., Chance, B. et al. Chemiluminescent measurement of increased free radical formation after ischemia/reperfusion. Digest Dis Sci 40, 1045–1053 (1995). https://doi.org/10.1007/BF02064197

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  • DOI: https://doi.org/10.1007/BF02064197

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