title: Alterations in zinc homeostasis underlie endothelial cell death induced by oxidative stress from acute exposure to hydrogen peroxide.
abstract: Oxidative stress has been associated with multiple pathologies and disease states, including those involving the cardiovascular system. Previously, we showed that pulmonary artery endothelial cells (PAECs) undergo apoptosis after acute exposure to H(2)O(2). However, the underlying mechanisms regulating this process remain unclear. Because of the prevalence of H(2)O(2) in normal physiological processes and apparent loss of regulation in disease states, the purpose of this study was to develop a more complete understanding of H(2)O(2)-mediated adverse effects on endothelial cell survival. Acute exposure of PAECs to H(2)O(2) caused a dose-dependent increase in cellular release of lactate dehydrogenase and a significant increase in production of superoxide ions, which appear to be generated within the mitochondria, as well as a significant loss of mitochondrial membrane potential and activity. Subsequent to the loss of mitochondrial membrane potential, PAECs exhibited significant caspase activation and apoptotic nuclei. We also observed a significant increase in intracellular free Zn(2+) after bolus exposure to H(2)O(2). To determine whether this increase in Zn(2+) was involved in the apoptotic pathway induced by acute H(2)O(2) exposure, we developed an adenoviral construct for overexpression of the Zn(2+)-binding protein metallothionein-1. Our data indicate that chelating Zn(2+), either pharmacologically with N,N,N',N-tetrakis(2-pyridylmethyl)ethylene diamine or by overexpression of the Zn(2+)-binding protein metallothionein-1, in PAECs conferred significant protection from induction of apoptosis and cell death associated with the effects of acute H(2)O(2) exposure. Our results show that the acute toxicity profile of H(2)O(2) can be attributed, at least in part, to liberation of Zn(2+) within PAECs. We speculate that regulation of Zn(2+) levels may represent a potential therapeutic target for cardiovascular disease associated with acute oxidative stress.
authors.affiliation: Vascular Biology Center, Medical College of Georgia, 1459 Laney Walker Blvd., Augusta, GA 30912, USA.
authors.forename: Dean A
authors.initials: DA
authors.lastname: Wiseman
authors.forename: Sandra M
authors.initials: SM
authors.lastname: Wells
authors.forename: Maryann
authors.initials: M
authors.lastname: Hubbard
authors.forename: Jonathan E
authors.initials: JE
authors.lastname: Welker
authors.forename: Stephen M
authors.initials: SM
authors.lastname: Black
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D002478:Cells, Cultured; D002614:Chelating Agents / Q000494:pharmacology; D042783:Endothelial Cells / Q000187:drug effects / Q000378:metabolism* / Q000473:pathology*; D005029:Ethylenediamines / Q000494:pharmacology; D006706:Homeostasis; D006861:Hydrogen Peroxide / Q000633:toxicity*; D008668:Metallothionein / Q000235:genetics / Q000378:metabolism; D018384:Oxidative Stress; D011651:Pulmonary Artery / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D012756:Sheep; D015398:Signal Transduction / Q000187:drug effects; D015032:Zinc / Q000378:metabolism*
chemical_list: D002614:Chelating Agents; D005029:Ethylenediamines; D011994:Recombinant Proteins; D008668:Metallothionein; D006861:Hydrogen Peroxide; D015032:Zinc; C044387:N,N,N',N'-tetrakis(2-pyridylmethyl)ethylenediamine