Effects of superoxide anions on endothelial Ca2+ signaling pathways.
Although the involvement of free radicals in the development of endothelial dysfunction under pathological conditions, like diabetes and hypercholesterolemia, has been proposed frequently, there is limited knowledge as to how superoxide anions (O2-) might affect endothelial signal transduction. In this study, we investigated the effects of preincubation with the O2(-)-generating system xanthine oxidase/hypoxanthine (XO/HX) on mechanisms for Ca2+ signaling in cultured porcine aortic endothelial cells. Incubation of cells with XO/HX yielded increased intracellular Ca2+ release and capacitative Ca2+ entry in response to bradykinin and ATP in a time- and concentration-dependent manner. This effect was prevented by superoxide dismutase but not by the tyrosine kinase inhibitor tyrphostin A48. In addition, capacitative Ca2+ entry induced by the receptor-independent stimulus 2,5-di-(tert-butyl)-1,4-benzohydroquinone or thapsigargin was enhanced in O2(-)-exposed cells (+38% and +32%, respectively). Increased Ca2+ release in response to bradykinin in XO/HX-pretreated cells might be due to enhanced formation of inositol-1,4,5-trisphosphate (+140%). Exposure to XO/HX also affected other signal transduction mechanisms involved in endothelial Ca2+ signaling, such as microsomal cytochrome P450 epoxygenase and membrane hyperpolarization to Ca2+ store depletion with thapsigargin (+103% and +48%, respectively) and tyrosine kinase activity (+97%). A comparison of bradykinin-initiated intracellular Ca2+ release and thapsigargin-induced hyperpolarization with membrane viscosity modulated by XO/HX (decrease in viscosity) or cholesterol (increase in viscosity) reflected a negative correlation between bradykinin-initiated Ca2+ release and membrane viscosity. Because intracellular Ca2+ is a main regulator of endothelial vascular function, our data suggest that O2- anions are involved in regulation of the vascular endothelium.
Graier
W F
WF
Department of Medical Biochemistry, University of Graz, Austria. wolfgang.graier@kfunigraz.ac.at
Hoebel
B G
BG
Paltauf-Doburzynska
J
J
Kostner
G M
GM
D000255:Adenosine Triphosphate / Q000494:pharmacology; D000818:Animals; D000838:Anions; D001011:Aorta; D001920:Bradykinin / Q000494:pharmacology; D002118:Calcium / Q000378:metabolism*; D002462:Cell Membrane / Q000502:physiology; D002478:Cells, Cultured; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism; D004730:Endothelium, Vascular / Q000187:drug effects / Q000378:metabolism* / Q000648:ultrastructure; D019271:Hypoxanthine / Q000378:metabolism; D007295:Inositol Phosphates / Q000378:metabolism; D008564:Membrane Potentials; D008861:Microsomes / Q000201:enzymology; D011505:Protein-Tyrosine Kinases / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects*; D013481:Superoxides / Q000378:metabolism / Q000494:pharmacology*; D013552:Swine; D014783:Viscosity; D014969:Xanthine Oxidase / Q000378:metabolism
D000838:Anions; D007295:Inositol Phosphates; D013481:Superoxides; D019271:Hypoxanthine; D000255:Adenosine Triphosphate; D003577:Cytochrome P-450 Enzyme System; D014969:Xanthine Oxidase; D011505:Protein-Tyrosine Kinases; D001920:Bradykinin; D002118:Calcium