Potential Role of the Endocannabinoid, Anandamide, in Reducing Cardiomyocyte Mitochondrial Function Following Alcohol Binge

Binge alcohol drinking is a growing health problem in the United States with an increasing number of emergency room visits. We recently demonstrated the involvement of the endocannabinoid (EC), anandamide (AEA), and its receptor, cannabinoid type 1 receptor (CB1R), in acute alcohol-induced cardiac dysfunction. Activation of CB1 receptors in the myocardium following binge alcohol drinking negatively affects cardiovascular function, impacting both cardiac contractility and vascular tone. However, it is unknown if the alcohol-induced increase in AEA levels in the myocardium leads to mitochondrial dysfunction that may contribute to impaired contractility in cardiomyocytes. We hypothesize that acute alcohol-induced AEA synthesis decreases cardiomyocyte mitochondrial bioenergetics and reduces cardiac performance through CB1R-dependent mechanisms. In this study, we used multidisciplinary approaches, including mass spectrometry, histological approaches, RNA sequencing, and seahorse mitochondrial analysis, to examine how the increased levels of AEA in the myocardium alter cardiac mitochondrial function. Utilizing a mouse model of acute alcohol intoxication, we found that oral administration of alcohol (5g/kg) leads to an increase in cardiac AEA levels that was paralleled with cardiac dysfunction and the tissue accumulation of 4-hydroxynonenal, a stable byproduct of lipid peroxidation. RNA sequencing analysis of cardiac tissue showed a significant increase in the expression of genes related to AEA synthesis (Abhd4, Gde-1) in hearts three hours after alcohol administration compared to control samples. Additionally, these genes showed a strong positive correlation with other differentially expressed genes implicated in mitochondrial homeostasis and regulation of cellular bioenergetics. To assess the direct effects of AEA on cardiac mitochondrial function, neonatal mouse cardiomyocytes (NMCMs) were isolated from pups at 3 days of age. The cells were cultured for 6 days and then treated with AEA at 1nM, 10nM, 100nM, and 1µM for one hour immediately before measuring cellular respiration using the Seahorse XF pro bioanalyzer. AEA decreased maximal respiration, spare respiratory capacity, and ATP production, indicating there are negative effects of AEA on cardiomyocyte mitochondrial function and cellular bioenergetics. These findings demonstrate the potential role of anandamide in the alteration of cardiac mitochondrial function following alcohol binge drinking, further impacting cardiac contractility. Supported by T32AA007577-2, R00AA028300, and LSUHSC start-up funds This abstract was presented at the American Physiology Summit 2025 and is only available in HTML format. There is no downloadable file or PDF version. The Physiology editorial board was not involved in the peer review process.