Endothelium-derived 2-arachidonylglycerol: an intermediate in vasodilatory eicosanoid release in bovine coronary arteries

• Published in: American journal of physiology. Heart and circulatory physiology Vol. 288; no. 3; pp. H1344 - H1351
• Main Authors: Gauthier, Kathryn M, Baewer, David V, Hittner, Sarah, Hillard, Cecilia J, Nithipatikom, Kasem, Reddy, D. Sudarshan, Falck, J. R, Campbell, William B
• Format: Journal Article
• Language: English
• Published: United States 01.03.2005
• Subjects: 8,11,14-Eicosatrienoic Acid - analogs & derivatives; 8,11,14-Eicosatrienoic Acid - metabolism; Animals; Arachidonic Acid - pharmacokinetics; Arachidonic Acids - metabolism; Carbon Radioisotopes; Cattle; Cells, Cultured; Coronary Vessels - cytology; Coronary Vessels - physiology; Endocannabinoids; Endothelium, Vascular - cytology; Endothelium, Vascular - metabolism; Glycerides - metabolism; Hydroxyeicosatetraenoic Acids - metabolism; Vasodilation - physiology
• ISSN: 0363-6135; 1522-1539
• DOI: 10.1152/ajpheart.00537.2004

1 Department of Pharmacology and Toxicology, Medical College of Wisconsin, Milwaukee, Wisconsin; and 2 Department of Biochemistry, University of Texas Southwestern Medical Center, Dallas, Texas Submitted 10 June 2004 ; accepted in final form 1 November 2004 Acetylcholine stimulates the release of endothelium-derived arachidonic acid (AA) metabolites including prostacyclin and epoxyeicosatrienoic acids (EETs), which relax coronary arteries. However, mechanisms of endothelial cell (EC) AA activation remain undefined. We propose that 2-arachidonylglycerol (2-AG) plays an important role in this pathway. An AA metabolite isolated from bovine coronary ECs was identified as 2-AG by mass spectrometry. In ECs pretreated with the fatty acid amidohydrolase inhibitor diazomethylarachidonyl ketone (DAK; 20 µmol/l), methacholine (10 µmol/l)-stimulated 2-AG release was blocked by the phospholipase C inhibitor U-73122 (10 µmol/l) or the diacylglycerol lipase inhibitor RHC-80267 (40 µmol/l). In U-46619-preconstricted bovine coronary arterial rings, 2-AG relaxations averaging 100% at 10 µmol/l were inhibited by endothelium removal, by DAK, by the hydrolase inhibitor methyl arachidonylfluorophosphate (10 µmol/l), by the cyclooxygenase inhibitor indomethacin (10 µmol/l), but not by the CB1 cannabinoid receptor antagonist SR-141716 (1 µmol/l). The cytochrome P -450 inhibitor SKF-525a (10 µmol/l) and the 14,15-epoxyeicosa-5 Z -enoic acid EET antagonist (14,15-EEZE; 10 µmol/l) further attenuated the indomethacin-resistant relaxations. The nonhydrolyzable 2-AG analogs noladin ether, 2-AG amide, and 14,15-EET glycerol amide did not induce relaxation. N -nitro- L -arginine-resistant relaxations to methacholine were also inhibited by U-73122, RHC-80267, and DAK. 14,15-EET glycerol ester increased opening of large-conductance K + channels 12-fold in cell-attached patches of isolated smooth muscle cells and induced relaxations averaging 95%. These results suggest that methacholine stimulates EC 2-AG production through phospholipase C and diacylglycerol lipase activation. 2-AG is further hydrolyzed to AA, which is metabolized to vasoactive eicosanoids. These studies reveal a role for 2-AG in EC AA release and the regulation of coronary tone. arachidonic acid; epoxyeicosatrienoic acids; vascular relaxation; prostacyclin; endothelium-derived hyperpolarizing factor Address for reprint requests and other correspondence: K. M. Gauthier, Dept. of Pharmacology and Toxicology, Medical College of Wisconsin, 8701 Watertown Plank Rd., Milwaukee, WI 53226 (E-mail: kgauth{at}mcw.edu )