Anandamide and cyclooxygenase-2 regulate the interaction between endometrial fibroblast and endothelial cells.

CAÑUMIL VA; BELTRAME JS; BOGETTI M; SCOTTI L; FRANCHI A; PARBORELL MF; MERESMAN G; RIBEIRO ML

Congreso; XXIII Reunión Anual de la Sociedad Argentina de Biología (SAB); 2021

During pregnancy, an adequate uterine blood flow is required to ensure the formation of the placenta and the delivery of oxygen and nutrients to the developing embryo. This change in blood flow is the result of the process of angiogenesis and remodeling of the maternal vessels. The transformation of these vessels requires a dynamic interaction between different cell types of the maternal–fetal interface, such as trophoblast cells, endometrial fibroblasts, and endothelial cells, among others. Anandamide (AEA) is a lipid molecule from the group of endocannabinoids. AEA shows pro-implantation characteristics and participates in the processes that take place at the maternal–fetal interface during implantation. Furthermore, AEA has been shown to regulate cyclooxygenase-2 (COX-2) pathway, and that COX-2-derived prostaglandins play a crucial role at implantation sites. However, the role of AEA during vascular remodeling of the maternal–fetal interface has not been studied. Therefore, the first objective of the present work was to investigate the effect of AEA on the vascular behavior of endometrial stromal fibroblasts (wound healing assay). For this, T-hESC cells (cell line derived from human endometrial stromal fibroblasts) were seeded in 24-well plates until confluence. The wound was made, and cells were incubated with 1 nM, 10 nM, 100 nM, or 1000 nM AEA for 12 h. The percentage of wound closure was measured as [(T0h cell-free area − T12h cell-free area) × 100]/ T0h area. Once the effective concentration of AEA was determined, the participation of COX-2 was evaluated, using a selective COX-2 inhibitor (meloxicam). Subsequently, the effect of AEA in the interaction between endometrial stromal fibroblasts and maternal endothelial cells was evaluated. The supernatants from T-hESC migration assay performed in the presence of AEA were collected, and used as conditioned media in endothelial cell migration (EA.hy926). First, we observed that AEA stimulated the migration of T-hESC cells in a concentration-dependent manner, and COX-2 mediated this effect. On the other hand, the conditioned media from the migration of T-hESC cells incubated with AEA stimulated the migration of EA.hy926 cells. We propose that AEA stimulates the release of solublefactors derived from the COX-2 pathway that regulate the migration of stromal fibroblasts, as well as the interaction between this cell type and the endothelium. Taken together, our results suggest the participation of AEA in the vascular remodeling that takes place in the uterus during early gestation by a mechanism that involves the COX-2 isoform.