title: Peroxynitrite treatment reduces adenosine uptake via the equilibrative nucleoside transporter in rat astrocytes.
abstract: In the oxidative stress-loaded brain, extracellular adenosine levels are elevated and thereby neuronal damage is attenuated, but mechanisms underlying alteration of the extracellular kinetics of adenosine remain unclear. Here we investigated whether oxidative stress might alter functional expression of nucleoside transporters (NTs), a predominant regulatory system for nucleoside kinetics, in cultured rat astrocytes. Treatment of astrocytes with 0.5mM SIN-1 for 3h caused apparent cellular accumulation of nitrotyrosine, but had no effect on their viability, indicating load of oxidative stress to astrocytes without any change in their viability. Under the condition, [(3)H]adenosine uptake was significantly less than that by control cells. This decreased uptake was due to decrease in adenosine uptake mediated by an equilibrative NT (ENT) 1 which was inhibited by low concentrations (≤0.1 μM) of nitrobenzylthioinosine (NBMPR), but not by sodium-dependent or high concentrations (≥1 μM) of NBMPR-inhibitable nucleoside transporters. The expression level of ENT1 was not altered, while the Michaelis constant, but not the maximum rate, of adenosine uptake was increased. These findings suggest that under oxidative stress-loaded conditions, decreased adenosine clearance via astrocytic ENT1 might involve, at least in part, in an elevated extracellular adenosine level in the brain.
authors.affiliation: Department of Environmental Biochemistry, Kyoto Pharmaceutical University, 5 Nakauchi-cho, Misasagi, Yamashina-ku, Kyoto 607-8414, Japan.
authors.forename: Akiko
authors.initials: A
authors.lastname: Tanaka
authors.forename: Kentaro
authors.initials: K
authors.lastname: Nishida
authors.forename: Hiroto
authors.initials: H
authors.lastname: Okuda
authors.forename: Takeshi
authors.initials: T
authors.lastname: Nishiura
authors.forename: Youichirou
authors.initials: Y
authors.lastname: Higashi
authors.forename: Sadaki
authors.initials: S
authors.lastname: Fujimoto
authors.forename: Kazuki
authors.initials: K
authors.lastname: Nagasawa
mesh_terms: D000241:Adenosine / Q000378:metabolism*; D000818:Animals; D001253:Astrocytes / Q000187:drug effects / Q000378:metabolism*; D015153:Blotting, Western; D007150:Immunohistochemistry; D008981:Molsidomine / Q000031:analogs & derivatives / Q000494:pharmacology; D020030:Nitric Oxide Donors / Q000494:pharmacology; D033703:Nucleoside Transport Proteins / Q000187:drug effects / Q000378:metabolism*; D016877:Oxidants / Q000378:metabolism / Q000494:pharmacology; D018384:Oxidative Stress / Q000502:physiology*; D030421:Peroxynitrous Acid / Q000378:metabolism / Q000494:pharmacology; D051381:Rats; D017208:Rats, Wistar
chemical_list: D020030:Nitric Oxide Donors; D033703:Nucleoside Transport Proteins; D016877:Oxidants; D030421:Peroxynitrous Acid; C002385:linsidomine; D008981:Molsidomine; D000241:Adenosine