Potential role of mitochondrial calcium metabolism during reperfusion injury

A. A. Vlessis, L. Mela-Riker

Research output: Contribution to journalArticle

20 Scopus citations

Abstract

Ischemia-reperfusion injury has been associated with intracellular H2O2 and superoxide radical production from accumulated hypoxanthine (HX) and xanthine oxidase (XO). The effect of H2O2 and superoxide radical on mitochondrial Ca2+ efflux was characterized in isolated renal mitochondria using a HX-XO system. Mitochondria were suspended in buffered medium containing 200 μM HX. Extramitochondrial Ca2+ was monitored kinetically at 660-685 nm using the Ca2+ indicator arsenazo III. After preloading mitochondria with 18-25 nmol Ca2+/mg protein, addition of XO to the medium caused a rapid oxidation of mitochondrial NAD(P)H followed by Ca2+ release. Ca2+ efflux was attributed to mitochondrial metabolism of H2O2 because efflux could be prevented with catalase but not superoxide dismutase. The Ca2+ efflux rate (r = 0.995) and lag time to Ca2+ efflux (r = 0.987) both correlate well with the NAD(P)H oxidation rate. Exogenous ATP prevents Ca2+ efflux in a dose-dependent fashion (K(m) = 35 μM ATP) without affecting NAD(P)H oxidation; ATP plus oligomycin, however, had no effect. The protective effect of ATP on Ca2+ efflux was diminished by ruthenium red (RR). XO-induced Ca2+ efflux increased state 4 respiration 148% via a futile Ca2+ cycle involving the Ca2+ uniport. The increase in state 4 respiration could be reversed with RR (α < 0.001) or ATP (α < 0.01); ATP plus oligomycin, however, had no efect. The results are discussed in relation to the oxygen free radical theory of reperfusion injury.

Original languageEnglish (US)
Pages (from-to)25/6
JournalAmerican Journal of Physiology - Cell Physiology
Volume256
Issue number6
StatePublished - Jan 1 1989

ASJC Scopus subject areas

  • Physiology
  • Cell Biology

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