Anandamide (AEA) is involved in decidual nitric oxide (NO) production induced by lipopolysaccharide (LPS) on early murine pregnancy.

VERCELLI C; AISEMBERG J; BILLI S; CERVINI M; RIBEIRO ML; FRANCHI AM

Saint-Sauveur, Québec, Canadá

Simposio; 17th Annual Symposium on the Cannabinoids. International Cannabinoid Research Society (ICRS).; 2007

International Cannabinoid Research Society

Nitric oxide (NO) fulfils important functions during pregnancy and has a role in implantation, decidualization, vasodilatation and myometrial relaxation. However, at high concentrations, such as those that are produced in sepsis, NO has toxic effects as it is a free radical. Our previous results indicate that LPS, an integral part of the outer membrane of Gram negative bacteria, is capable of producing embryonic resorption in mice due to NO increased production not only in uterus but also in decidua. Recent research has revealed that LPS induces AEA synthesis in murine macrophages. The aim of the present work was to determine the effect of LPS and AEA on decidual NO-derived reactive species production induced by LPS on early murine pregnancy. On day 7 of pregnancy female mice were sacrificed by cervical dislocation and uterus and decidua were separated in each implantation site. Decidua was then incubated for 12 h in the presence of a) LPS (1 ug/ml), b) AEA (10-8M), c) LPS + AM251 (10-9M) (cannabinoid type 1 (CB1) receptor antagonist) and d) LPS + SR144528 (10-8M) (cannabinoid type 2 (CB2) receptor antagonist) and NO was measured in culture supernatants as NO3- plus NO2-. Both LPS and AEA were capable of increasing NO levels and NO production induced by LPS was inhibited when decidua was incubated in the presence of AM251 as well as in the presence of SR144528. We also determine protein nitration by western blot in decidua incubated for 12 h in the presence/absence of LPS. We observed an increase in tyrosine nitration in a time dependent manner.                 We determined the activity of the fatty acid amide hydrolase (FAAH–the enzyme responsible for hydrolysis of AEA- (conversion of [3H]-anandamide in [14C]-arachidonic acid and ethanolamine)) in the presence/absence of LPS. Treatment with the endotoxin increased FAAH activity.                 These results and the inhibition of LPS-induced augmentation of NO synthesis by both cannabinoid receptor antagonists suggest that AEA could be an intermediate in LPS effect.