Intracellular Ca2+ homeostatic regulation and 4-hydroxynonenal-induced aortic endothelial dysfunction.
The aldehydic lipid peroxidation product 4-hydroxynonenal (HNE) is known to compromise erythrocyte passive Ca2+ permeability and to irreversibly inhibit the plasma membrane (Ca2+ + Mg2+)-ATPase and Ca2+-transport. To measure the effects of HNE on passive and active Ca2+ transport in endothelial cells, we first characterized 45Ca2+ uptake and efflux in cultured porcine aortic endothelial cells (PAEC). PAEC exchanged 45Ca2+ to a cumulative near-isotopic equilibrium of about 4.5 pmole 45Ca2+/10(6) cells in 120 min at 37 degrees C. This Ca2+ pool was diminished by thapsigargin, cyclopiazonic acid, oligomycin B, and sodium azide. In contrast, ouabain enhanced Ca2+ uptake capacity from 5.17 to 5.77 pmole/10(6) cells. Accumulated 45Ca2+ was extruded at rate of 8.7 fmole 45Ca2+/10(6) cells/min or shunted rapidly by the ionophore A23187. HNE increased total 45Ca2+ accumulation in a time- and concentration-dependent manner by as much as 562% with an EC50 of 64.0 wM. Concomitant morphological analysis of PAEC revealed vacuolization, nuclear swelling, cell shrinking, and cell detachment. Initial structural changes, such as vacuolization, began well before any changes in Ca2+ accumulation were observed. These functional and morphological changes indicate that HNE significantly increases intracellular Ca2+ accumulation in vascular endothelium, which may explain the cytotoxic effects associated with HNE exposure and provide further evidence that atherogenic effects of HNE may, in part, be caused by disturbances in Ca2+ homeostasis.
McConnell
Elizabeth J
EJ
Department of Pharmacology and Toxicology, Indiana University School of Medicine, Evansville, IN 47712, USA.
Raess
Beat U
BU
D000447:Aldehydes / Q000378:metabolism* / Q000494:pharmacology; D000818:Animals; D001013:Aorta, Thoracic / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D002118:Calcium / Q000378:metabolism*; D002478:Cells, Cultured; D004305:Dose-Response Relationship, Drug; D004730:Endothelium, Vascular / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D006706:Homeostasis; D007424:Intracellular Fluid / Q000378:metabolism; D017136:Ion Transport / Q000187:drug effects; D015227:Lipid Peroxidation; D013552:Swine
D000447:Aldehydes; C027576:4-hydroxy-2-nonenal; D002118:Calcium