Endocannabinoid System Regulation in Pyometra-Affected and Healthy Canine Uteri

Pyometra is a frequent and life-threatening reproductive disorder in bitches, characterized by profound immune and inflammatory responses within the uterus. The endocannabinoid system (eCS) is a key modulator of immune regulation, tissue homeostasis, and inflammation; however, its role in canine uterine physiology and pathology remains unexplored. This study aimed to characterize the presence and regulation of eCS components in the uterus of healthy and pyometra-affected dogs. Twenty-eight bitches were categorized into four groups: closed-cervix pyometra (CP; n=7), open-cervix pyometra (OP; n=7), diestrus (DE; n=7), and anestrus (AE; n=7). Uterine tissues were obtained by ovariohysterectomy. Serum progesterone (P4), anandamide (AEA), and 2-arachidonylglycerol (2-AG) concentrations were quantified, while uterine expression of cannabinoid receptors (CB1 and CB2) was assessed using real-time PCR and localized by immunohistochemistry (IHC). Serum AEA levels were significantly reduced in CP compared with AE (p=0.017), whereas 2-AG differences did not reach significance (p=0.072). Both CB1 and CB2 were consistently expressed across all groups, with IHC revealing receptor-specific patterns within uterine compartments. Collectively, these findings demonstrate for the first time that the canine uterus possesses a functional eCS, and that its modulation is linked to reproductive physiology and pyometra-associated inflammatory processes.