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

Mauro Maccarrone, Monica Bari, Tatiana Lorenzon, Tiziana Bisogno, Vincenzo Di Marzo, Alessandro Finazzi-Agrò

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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 α-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 α-linolenoylvanillyl-amide/N-(4-hydroxyphenyl)-arachidonoylamide were found to attenuate and enhance, respectively, this effect of AEA. Consistently, activation of CB1 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 HUVECs in a fashion sensitive to the NO synthase inhibitor Nω-nitro-L- arginine methyl ester. These findings suggest a NO-mediated regulatory loop between CB1 cannabinoid receptors and AEA transporter.

Original languageEnglish
Pages (from-to)13484-13492
Number of pages9
JournalJournal of Biological Chemistry
Issue number18
Publication statusPublished - May 5 2000


ASJC Scopus subject areas

  • Biochemistry

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