A physiological concentration of anandamide promotes the migration of human endometrial fibroblast and the interaction with endothelial cells in vitro

CAÑUMIL, VANESA A.; DE LA CRUZ BORTHIRY, FERNANDA L.; SCHEFFER, FRIDA; HERRERO, YAMILA; SCOTTI, LEOPOLDINA; BOGETTI, MARÍA EUGENIA; PARBORELL, FERNANDA; MERESMAN, GABRIELA F.; FRANCHI, ANA M.; BELTRAME, JIMENA S.; RIBEIRO, MARÍA L.

PLACENTA

W B SAUNDERS CO LTD

Año: 2023

0143-4004

Introduction: The mechanisms that govern fibroblast behavior during the vascular adaptations ofthe uterus at early pregnancy remain unknown. Anandamide, an endocannabinoid, binds tocannabinoid receptors (CBs), and regulates gestation and angiogenesis. Its tone is regulated byfatty acid amide hydrolase (FAAH) within the uterus. We investigated the role of anandamide inendometrial fibroblasts migration and whether anandamide modulates fibroblasts-endothelialcrosstalk.Methods: T-hESC and EA.hy926 cell lines were used as models of endometrial stromal andendothelial cells, respectively. T-hESC were incubated with anandamide plus different agents.Migration was tested (wound healing assay and phalloidin staining). Protein expression andlocalization was studied by Western blot and immunofluorescence. To test fibroblast-endothelialcrosstalk, EA.hy926 cells were incubated with fibroblast conditioned media obtained after T-hESCmigration.Results: Anandamide 1 nM increased T-hESC migration via CB1 and CB2. Cyclooxygenase-2participated in anandamide-stimulated fibroblast migration. Prostaglandin F2alpha, and notprostaglandin E2, increased fibroblast wound closure. CB1, CB2, cyclooxygenase-2 and FAAH wereexpressed in T-hESC. Anandamide did not alter cyclooxygenase-2 localization but induced itscytoplasmic and nuclear expression through CB1 and CB2. URB-597, a FAAH selective inhibitor,also increased T-hESC migration via both CBs, and augmented cyclooxygenase-2 expression.Conditioned media from anandamide-induced T-hESC wound healing closure stimulatedendothelial migration and did not alter their proliferation. Soluble factors from cyclooxygenase-2were secreted by T-hESC and participated in T-hESC-induced EA.hy926 migration. Althoughanandamide-conditioned media augmented in EA.hy926 the expression of γH2AX, a marker ofDNA damage, cyclooxygenase-2 was not involved in this effect.