Iron loading of endothelial cells augments oxidant damage

G. Balla, G. M. Vercellotti, J. W. Eaton, H. S. Jacob

Research output: Contribution to journalArticle

109 Citations (Scopus)

Abstract

Transition metals, particularly iron, will potentiate oxidant damage to isolated cell organelles, plasma membranes, and DNA when added to in vitro incubation systems. However, similar studies of intact cells have been hampered by the relative impermeability of whole cells to iron. We have iron loaded cultured endothelial cells by using the iron-chelating fungistat 8-hydroxyquinoline (8HQ). 8HQ forms lipophilic chelates with iron and rapidly transfers the metal across the intact plasma membrane of endothelial cells. After brief exposure to 8HQ and subsequent thorough washing of endothelial cells, the cell-associated iron cannot be removed by the powerful chelator deferoxamine, clearly indicating the intracelullar location of 8HQ-transported iron. Iron-loaded cells (but not cells exposed to high concentrations of 8HQ or iron separately) are extremely sensitive to oxidants (1) produced externally by phorbol-stimulated granulocytes, (2) generated intracellularly by menadione, or (3) added as H2O2. In the latter instance, as little as 7 μmol/L H2O2 provokes destruction of ~50% of iron-loaded endothelial cells, whereas untreated endothelium readily survives exposure to H2O2 concentrations as high as 2 mmol/L. Cytotoxicity is accompanied by membrane lipid peroxidation (formation of thiobarbituric) acid-reactive substances). Both cytotoxicity and lipid peroxidation are inhibited by the lipophilic 21-adminosteroid U74500A ('lazaroid') (50% inhibitory concentration = approximately 0.5 μmol/L), whereas deferoxamine (250 μmol/) is ineffective (suggesting iron intercalation into hydrophobic domains of the cell). We conclude that this pharmacologic model for iron loading of intact cells may yield valuable insights inot the pathogenic importance of intracellular iron in iron overload states, inflammation, and cellular injury.

Original languageEnglish
Pages (from-to)546-554
Number of pages9
JournalJournal of Laboratory and Clinical Medicine
Volume116
Issue number4
Publication statusPublished - 1990

Fingerprint

Endothelial cells
Oxidants
Iron
Endothelial Cells
Oxyquinoline
Deferoxamine
terconazole
Lipid Peroxidation
Cell membranes
Cytotoxicity
Metals
Cell Membrane
Iron Chelating Agents
Vitamin K 3
Iron Overload
Thiobarbituric Acid Reactive Substances
Membrane Lipids
Chelating Agents
Granulocytes
Organelles

ASJC Scopus subject areas

  • Medicine(all)
  • Pathology and Forensic Medicine

Cite this

Balla, G., Vercellotti, G. M., Eaton, J. W., & Jacob, H. S. (1990). Iron loading of endothelial cells augments oxidant damage. Journal of Laboratory and Clinical Medicine, 116(4), 546-554.

Iron loading of endothelial cells augments oxidant damage. / Balla, G.; Vercellotti, G. M.; Eaton, J. W.; Jacob, H. S.

In: Journal of Laboratory and Clinical Medicine, Vol. 116, No. 4, 1990, p. 546-554.

Research output: Contribution to journalArticle

Balla, G, Vercellotti, GM, Eaton, JW & Jacob, HS 1990, 'Iron loading of endothelial cells augments oxidant damage', Journal of Laboratory and Clinical Medicine, vol. 116, no. 4, pp. 546-554.
Balla, G. ; Vercellotti, G. M. ; Eaton, J. W. ; Jacob, H. S. / Iron loading of endothelial cells augments oxidant damage. In: Journal of Laboratory and Clinical Medicine. 1990 ; Vol. 116, No. 4. pp. 546-554.
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