Anandamide (AEA) has vasodilator activity, which can be terminated by cellular re-uptake and degradation. Here we investigated the presence and regulation of the AEA transporter in human umbelical vein endothelial cells (HUVECs), HUVECs take up AEA by facilitated transport (apparent K-m = 190 +/- 10 nM and V-max = 45 +/- 3 pmol min(-1) . mg(-1) protein), which is inhibited by alpha-linolenoyl-vanillyl-amide and N-(4-hydroxyphenyl) arachidonoylamide, and stimulated up to 2.2-fold by nitric oxide (NO) donors. The NO scavenger hydroxocobalamin abolishes the latter effect, which is instead enhanced by superoxide anions but inhibited by superoxide dismutase and N-acetylcysteine, a precursor of glutathione synthesis. Peroxynitrite (ONOO-) causes a 4-fold activation of AEA transport into cells. The HUVEC AEA transporter contributes to the termination of a typical type 1 cannabinoid receptor (CB1) -mediated action of AEA i.e. the inhibition of forskolin-stimulated adenylyl cyclase, because NO/ONOO- donors and alpha-linolenoyl-vanillyl-amide/N-(4-hydroxyphenyl)-arachidonoylamide were found to attenuate and enhance, respectively, this effect of AEA. Consistently, activation of CB, cannabinoid receptors by either AEA or the cannabinoid HU-210 caused a stimulation of HUVEC inducible NO synthase activity and expression up to 2.9- and 2.6-fold, respectively. Also these effects are regulated by the AEA transporter, HU-210 enhanced AEA uptake by HOVECs in a fashion sensitive to the NO synthase inhibitor N omega-nitro-L-arginine methyl ester, These findings suggest a NO-mediated regulatory loop between CB1 cannabinoid receptors and AEA transporter.

Anandamide uptake by human endothelial cells and its regulation by nitric oxide

Maccarrone M;
2000

Abstract

Anandamide (AEA) has vasodilator activity, which can be terminated by cellular re-uptake and degradation. Here we investigated the presence and regulation of the AEA transporter in human umbelical vein endothelial cells (HUVECs), HUVECs take up AEA by facilitated transport (apparent K-m = 190 +/- 10 nM and V-max = 45 +/- 3 pmol min(-1) . mg(-1) protein), which is inhibited by alpha-linolenoyl-vanillyl-amide and N-(4-hydroxyphenyl) arachidonoylamide, and stimulated up to 2.2-fold by nitric oxide (NO) donors. The NO scavenger hydroxocobalamin abolishes the latter effect, which is instead enhanced by superoxide anions but inhibited by superoxide dismutase and N-acetylcysteine, a precursor of glutathione synthesis. Peroxynitrite (ONOO-) causes a 4-fold activation of AEA transport into cells. The HUVEC AEA transporter contributes to the termination of a typical type 1 cannabinoid receptor (CB1) -mediated action of AEA i.e. the inhibition of forskolin-stimulated adenylyl cyclase, because NO/ONOO- donors and alpha-linolenoyl-vanillyl-amide/N-(4-hydroxyphenyl)-arachidonoylamide were found to attenuate and enhance, respectively, this effect of AEA. Consistently, activation of CB, cannabinoid receptors by either AEA or the cannabinoid HU-210 caused a stimulation of HUVEC inducible NO synthase activity and expression up to 2.9- and 2.6-fold, respectively. Also these effects are regulated by the AEA transporter, HU-210 enhanced AEA uptake by HOVECs in a fashion sensitive to the NO synthase inhibitor N omega-nitro-L-arginine methyl ester, These findings suggest a NO-mediated regulatory loop between CB1 cannabinoid receptors and AEA transporter.
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11697/155835
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