Production of reactive oxygen by mitochondria from normoxic and hypoxic rat heart tissue

Thomas Paraidathathu, Herbert de Groot, James P. Kehrer

Research output: Contribution to journalArticle

75 Citations (Scopus)

Abstract

Reactive oxygen species (ROS), which may be involved in ischemic or reperfusion heart injury, can be produced by mitochondria. Previous work indicated that coupled mitochondria from ischemic heart tissue incubated in calcium-free medium produced less ROS than normal. The effects of calcium, which may be elevated in hypoxic or ischemic tissue, were not examined. The relative production of ROS by mitochondria from normoxic or hypoxic rat heart tissue was estimated by measuring the oxidation of dichlorofluorescin to the fluorescent compound, dichlorofluorescein. ROS were detectable during succinate-stimulated State 4 respiration. In the absence of calcium, mitochondria from hypoxic (60 min) heart tissue produced less ROS than mitochondria from normoxic heart tissue. In the presence of 0.1, 1, or 10 μM calcium, ROS produced by hypoxic mitochondria were increased to normoxic levels. While function was depressed in mitochondria from hypoxic tissue, the presence of 0.1 and 1 μM calcium had no further effect. Respiration was uncoupled in the presence of 10 μM calcium in mitochondria from both normoxic and hypoxic heart tissue. ROS production was increased in mitochondria from hypoxic tissue with both increasing concentrations of calcium and increasing duration of exposure. ROS production in mitochondria from normoxic heart tissue was only stimulated after 200 or more seconds of exposure to 1 or 10 μM calcium. Production of ROS in mitochondria from hypoxic tissue in the presence of 1 μM calcium was inhibited by rotenone (80%), ruthenium red (69%), and a combination of these agents (96%). In contrast, ruthenium red had no effect on ROS production by mitochondria from normoxic heart tissue. These data indicate that calcium can increase ROS production in mitochondria from hypoxic heart tissue. The production of ROS may be mediated by a reverse flow of electrons in the electron transport chain. The difference in the effect of calcium and ruthenium red on ROS production in mitochondria from normoxic and hypoxic tissue suggests that two different mechanisms may be involved.

Original languageEnglish
Pages (from-to)289-297
Number of pages9
JournalFree Radical Biology and Medicine
Volume13
Issue number4
DOIs
Publication statusPublished - 1992
Externally publishedYes

Fingerprint

Mitochondria
Rats
Reactive Oxygen Species
Tissue
Oxygen
Calcium
Heart Mitochondria
Ruthenium Red
Respiration
Heart Injuries
Rotenone
Succinic Acid
Electron Transport
Reperfusion Injury

Keywords

  • ardiac mitochondria
  • Calcium
  • Dichlorofluorescin diacetate
  • Free radicals
  • Heart
  • Hypoxia
  • Reactive oxygen
  • Succinate

ASJC Scopus subject areas

  • Clinical Biochemistry
  • Medicine(all)
  • Toxicology

Cite this

Production of reactive oxygen by mitochondria from normoxic and hypoxic rat heart tissue. / Paraidathathu, Thomas; de Groot, Herbert; Kehrer, James P.

In: Free Radical Biology and Medicine, Vol. 13, No. 4, 1992, p. 289-297.

Research output: Contribution to journalArticle

Paraidathathu, Thomas ; de Groot, Herbert ; Kehrer, James P. / Production of reactive oxygen by mitochondria from normoxic and hypoxic rat heart tissue. In: Free Radical Biology and Medicine. 1992 ; Vol. 13, No. 4. pp. 289-297.
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