Metabolism of anandamide in cerebral microvascular endothelial cells

Ping Chen, Shanming Hu, Shawn D. Harmon, Steven A. Moore, Arthur A. Spector, Xiang Fang

Research output: Contribution to journalArticle

9 Scopus citations

Abstract

Anandamide (N-arachidonoylethanolamine, AEA), an endogenous cannabinoid receptor agonist, causes potent vasodilation in the cerebral circulation through an endothelial-dependent or -independent mechanism. We have investigated the processing of [3H]AEA in cultured mouse cerebral microvascular endothelial cells (MEC) in order to better understand its mechanism of action in the cerebral vasculature. These cells took up anandamide very quickly, reaching a maximum value in 5min and remaining at that level for at least 8h. Analysis of the cell lipids demonstrated that, in addition to free anandamide, radioactivity was incorporated into phosphatidylcholine (PC), phosphatidylinositol (PI), and phosphatidylethanolamine (PE) in a time-dependent manner. Analysis of the hydrolyzed cell lipids indicated that anandamide was converted to arachidonic acid, a process that was inhibited by the selective fatty acid amide hydrolase inhibitor oleyl trifluoromethyl ketone (OTMK). Phospholipase A2 (PLA2) hydrolysis of the PC, PI, and PE fractions indicated that the arachidonic acid formed from anandamide was esterified predominately into sn-2 position of the endothelial phospholipids. Furthermore, anandamide and arachidonic acid were released when the cells were incubated with A23187. These results suggest that the biological activity of anandamide might be regulated by its rapid uptake and calcium-dependent release in endothelial cells, and conversion of anandamide to arachidonic acid might serve as an inactivation process in the cerebral microcirculation.

Original languageEnglish (US)
Pages (from-to)59-72
Number of pages14
JournalProstaglandins and Other Lipid Mediators
Volume73
Issue number1-2
DOIs
StatePublished - Jan 2004
Externally publishedYes

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Keywords

  • Amide hydrolase
  • Anandamide
  • Arachidonic acid
  • Endothelial cells
  • Phospholipids

ASJC Scopus subject areas

  • Biochemistry
  • Endocrinology

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