title: Reactive oxygen species-induced aortic vasoconstriction and deterioration of functional integrity.
abstract: Oxygen derived free radicals and other reactive oxygen species (ROS) are involved in a variety of disease states, which can have cardiac and vascular implications. The present study was performed to investigate the mechanism of ROS-induced vasoconstriction and the influence of ROS on the functional integrity of isolated rat thoracic aorta. ROS were generated by means of electrolysis (30 mA, during 0.5, 1, 2 or 3 min) of the organ bath fluid. ROS induced a transient (approximately 60 min) vasoconstriction and the maximally induced contraction was dependent on the duration of electrolysis. Dimethyl sulfoxide (DMSO) diminished the ROS-induced vasoconstriction almost completely, indicating a major influence of hydroxyl radicals on contractility. The dual cyclooxygenase/lipoxygenase inhibitor, meclofenamate, completely prevented the ROS-induced vasoconstriction. The phospholipase A2 (PLA2) inhibitor, oleyloxyethyl phosphorylcholine, was able to reduce the vasoconstriction elicited by ROS by approximately 70%. Conversely, the specific cytoplasmic PLA2 inhibitor arachidonyl trifluoromethylketone proved ineffective in this respect. By using the specific mitogen-activated protein kinase (MAPkinase) kinase inhibitor PD98059, it was shown that the activation of extracellular-regulated kinase (ERK) MAPkinase contributes to the ROS-induced vasoconstriction. The effects of ROS on the functional integrity of the aortae were investigated, in particular with respect to receptor (alpha1-adrenoceptor) and non-receptor-mediated contractile responses (high potassium solution). In addition, both the endothelium dependent (methacholine) and endothelium independent (sodium nitroprusside) vasorelaxation were investigated before and after ROS exposure. Electrolysis periods of 0.5 and 1 min induced a modest leftward shift of the concentration response curves for the alpha1-adrenoceptor agonist methoxamine. Longer electrolysis periods of 2 and 3 min additionally decreased the maximal response to (alpha1-adrenoceptor stimulation. Methacholine-induced vasorelaxation proved diminished in aortae subjected to electrolysis (0.5, 1, 2 and 3 min), whereas relaxation to sodium nitroprusside was nearly complete in all groups. KCl-induced contractions proved attenuated only after longer electrolysis periods of 2 and 3 min. This ROS-induced deterioration of functional integrity was almost completely prevented by 0.6% DMSO. From these results we may conclude that ROS induce an eicosanoid and ERK MAPkinase-mediated vasoconstriction in isolated rat thoracic aorta. In addition, exposure to ROS leads to a deterioration of functional integrity characterized by endothelial dysfunction and decreased contractile function.
authors.affiliation: Department of Pharmacotherapy, Academic Medical Center, University of Amsterdam, The Netherlands.
authors.forename: S L
authors.initials: SL
authors.lastname: Peters
authors.forename: M J
authors.initials: MJ
authors.lastname: Mathy
authors.forename: M
authors.initials: M
authors.lastname: Pfaffendorf
authors.forename: P A
authors.initials: PA
authors.lastname: van Zwieten
mesh_terms: D000818:Animals; D001013:Aorta, Thoracic / Q000187:drug effects* / Q000502:physiology; D019086:Bridged Bicyclo Compounds, Heterocyclic; D005231:Fatty Acids, Unsaturated; D005419:Flavonoids / Q000494:pharmacology; D006834:Hydrazines / Q000494:pharmacology; D007213:Indomethacin / Q000494:pharmacology; D008297:Male; D008729:Methoxamine / Q000494:pharmacology; D048052:Mitogen-Activated Protein Kinase 3; D020928:Mitogen-Activated Protein Kinases / Q000502:physiology; D011189:Potassium Chloride / Q000494:pharmacology; D051381:Rats; D017208:Rats, Wistar; D017382:Reactive Oxygen Species / Q000378:metabolism; D014661:Vasoconstriction / Q000187:drug effects*
chemical_list: D019086:Bridged Bicyclo Compounds, Heterocyclic; D005231:Fatty Acids, Unsaturated; D005419:Flavonoids; D006834:Hydrazines; D017382:Reactive Oxygen Species; D011189:Potassium Chloride; C045749:SQ 29548; D048052:Mitogen-Activated Protein Kinase 3; D020928:Mitogen-Activated Protein Kinases; D008729:Methoxamine; C093973:2-(2-amino-3-methoxyphenyl)-4H-1-benzopyran-4-one; D007213:Indomethacin