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• W2072561293 abstract "Abstract Hypoxia relates with altered placental vasodilation, and in isolated endothelial cells, it reduces activity of the endothelial nitric oxide synthase (eNOS) and l -arginine transport. It has been reported that arginase-2 expression, an alternative pathway for l -arginine metabolism, is increased in adult endothelial cells exposed to hypoxia as well as in pre-eclamptic placentae. We studied in human umbilical vein endothelial cells (HUVEC) whether hypoxia-reduced NO synthesis results from increased arginase-mediated l -arginine metabolism and changes in subcellular localization of eNOS and arginase-2. In HUVEC exposed (24 h) to 5% (normoxia) or 2% (hypoxia) oxygen, l -arginine transport kinetics, arginase activity (urea assay), and NO synthase (NOS) activity ( l -citrulline assay) were determined. Arginase-1, arginase-2 and eNOS expression were determined by RT-PCR and Western blot. Subcellular localization of arginase-2 and eNOS were studied using confocal microscopy and indirect immunofluorescence. Experiments were done in absence or presence of S -(2-boronoethyl)- l -cysteine-HCl (BEC, arginase inhibitor) or N G -nitro- l -arginine methyl ester ( l -NAME). Hypoxia-induced reduction in eNOS activity was associated with a reduction in eNOS phosphorylation at Serine-1177 and increased phosphorylation at Threonine-495. This was paralleled with an induction in arginase-2 expression and activity, and decreased l -arginine transport. In hypoxia the arginase inhibition, restored NO synthesis and l -arginine transport, without changes in the eNOS post-translational modification status. Hypoxia increased arginase-2/eNOS colocalization, and eNOS redistribution to the cell periphery. Altogether these data reinforce the thought that eNOS cell location, post-translational modification and substrate availability are important mechanisms regulating eNOS activity. If these mechanisms occur in pregnancy diseases where feto-placental oxygen levels are reduced remains to be clarified." @default.
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• W2072561293 date "2011-12-01" @default.
• W2072561293 modified "2023-10-17" @default.
• W2072561293 title "Hypoxia-reduced nitric oxide synthase activity is partially explained by higher arginase-2 activity and cellular redistribution in human umbilical vein endothelium" @default.
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• W2072561293 doi "https://doi.org/10.1016/j.placenta.2011.09.003" @default.
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