Endocannabinoid hydrolases differentially distribute in platelets and red blood cells and are differentially released by thrombin

Abstract

BackgroundPlasma levels of the major endocannabinoids 2-arachidonoylgycerol (2AG) and anandamide (N-arachidonoylethanolamine, AEA) have been identified to vary independently with particular pathological conditions. The levels of these endocannabinoids are tightly regulated by two hydrolytic enzymes, monoacylglycerol lipase (MAGL) and fatty acid amide hydrolase (FAAH), respectively. ObjectivesIn this study, we have quantified these enzyme activities in the major blood fractions. Patients/MethodsIn blood fractions from human volunteers, radiometric assays were used to quantify monoacylglycerol lipase and fatty acid amide hydrolase. Tagging with fluorophosphonate-rhodamine allowed quantification of platelet serine hydrolase activities. ResultsFatty acid amide hydrolase activity was highest in platelets, while MAGL activity was most abundant in erythrocytes. Sampling the blood of donors on two further occasions 15 days apart showed no significant change in platelet FAAH or erythrocyte MAGL activities. Activities were not different when comparing female donors with males. Storage of these blood fractions at − 80 °C was associated with a rapid loss in enzyme activities, which could largely by avoided by storage in liquid nitrogen. Incubation of platelets and erythrocytes in the presence of thrombin lead to release of measurable FAAH, but not MAGL, activity. Tagging of serine hydrolase activities with fluorophosphonate-rhodamine allowed confirmation of MAGL activity in platelet preparations, as well as multiple other enzymes. ConclusionsThese investigations suggest a potential role for FAAH in regulation of coagulation, while the role of MAGL in blood requires further investigation.