title: Nitrosothiol stores in vascular tissue: modulation by ultraviolet light, acetylcholine and ionomycin.
abstract: Our previous studies demonstrated that light-induced vascular relaxation (photorelaxation) was mediated by a tissue source of nitric oxide that was independent of endothelial nitric oxide synthase (eNOS), but sensitive to inhibitors of soluble guanylate cyclase, extracellular nitric oxide scavengers and possessed the properties of a nitrosothiol. In the present study we describe High Performance Liquid Chromatography and spectrofluorometric techniques that allowed us to measure tissue levels of the nitrosothiol, S-nitrosoglutathione and its modulation in mouse aortic tissues, smooth muscle cells and human umbilical vein endothelial cells (HUVECs) following exposure to exogenous S-nitrosoglutathione, light and chemical stimuli. Basal levels of S-nitrosoglutathione were similar in control mouse aortae and HUVECs and the store size could be enhanced by exposure of tissues/cells to nitric oxide solution. No basal S-nitrosoglutathione was detected in tissue from diabetic db/db mice; however, ultraviolet light was still able to elicit relaxation of aortic tissues. Ultraviolet light induced the release of nitric oxide from the S-nitrosoglutathione store with an associated increase in the concentration of nitrite. The release of nitric oxide from the store in HUVECs was modulated by extracellular oxidative stress induced by xanthine/xanthine oxidase and also, in an atropine-sensitive process, by acetylcholine, as well as by the calcium ionophore, ionomycin. These interventions resulted in a reduced S-nitrosoglutathione store and elevated levels of nitrite. These data suggest that endothelial and vascular smooth muscle cells possess stores of nitric oxide that, in part, exist in the form of S-nitrosoglutathione. Furthermore, these stores, albeit small, may provide an additional mechanism for the regulation of vascular tone, especially under conditions, such as diabetes, in which nitric oxide generation or bioavailability is compromised; however, additional studies are required to determine not only whether there are additional chemical storage forms of nitric oxide, but also the location of such stores.
authors.affiliation: Smooth Muscle Research Group, Department of Pharmacology and Therapeutics, Faculty of Medicine, University of Calgary, Alberta, Canada.
authors.forename: Ella S M
authors.initials: ES
authors.lastname: Ng
authors.forename: Zhong-Jian
authors.initials: ZJ
authors.lastname: Cheng
authors.forename: Anthie
authors.initials: A
authors.lastname: Ellis
authors.forename: Hong
authors.initials: H
authors.lastname: Ding
authors.forename: Yanfen
authors.initials: Y
authors.lastname: Jiang
authors.forename: Yang
authors.initials: Y
authors.lastname: Li
authors.forename: Morley D
authors.initials: MD
authors.lastname: Hollenberg
authors.forename: Chris R
authors.initials: CR
authors.lastname: Triggle
mesh_terms: D000109:Acetylcholine / Q000494:pharmacology*; D000818:Animals; D001011:Aorta / Q000187:drug effects / Q000378:metabolism / Q000528:radiation effects; D002478:Cells, Cultured; D042783:Endothelial Cells / Q000378:metabolism*; D006801:Humans; D015759:Ionomycin / Q000494:pharmacology*; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D009131:Muscle, Smooth, Vascular / Q000166:cytology / Q000378:metabolism*; D009569:Nitric Oxide / Q000378:metabolism; D026422:S-Nitrosoglutathione / Q000378:metabolism*; D014466:Ultraviolet Rays; D019820:Xanthine / Q000494:pharmacology
chemical_list: D019820:Xanthine; D009569:Nitric Oxide; D015759:Ionomycin; D026422:S-Nitrosoglutathione; D000109:Acetylcholine