Anandamide impairs the syncytialization of human cytotrophoblast cells

PALLIGRAS MARCOS; ABÁN CYNTIA; ETCHEVERRY, TOMÁS; VALEFF NATALÍN; MARTÍNEZ NORA; ACCIALINI PAULA; SARACO NORA; FARINA MARIANA

Mar del Plata

Congreso; LXIII Reunión Científica Anual de la Sociedad Argentina de Investigación Clínica (SAIC); 2018

The syncytiotrophoblast (STB) is the main structural and functional layer of the human placenta. The underlying cytotrophoblast cells (CTB) fuse to form this multinucleated syncytium in a process called syncytialization. Disturbance in this event is associated with pregnancy pathologies such as preeclampsia and intrauterine growth restriction.Endocannabinoids (eCBs) are a group of bioactive lipid mediators which, together with their receptors and the enzymes involved in their metabolism, constitute the endocannabinoid system (ECS). Anandamide (N-arachidonylethanoalmine, AEA) and 2-Arachidonoylglycerol (2-AG) are the major eCBs, having both important roles in human placentation. Recently it has been reported that 2-AG impairs CTB fusion. However, the role of AEA in this process remains unknown.The aim of this work was to study the impact of AEA on syncytialization and to elucidate the mechanisms involved in this process.BeWo cell line was cultured with 25µM forskolin (FSK) to induce syncytialization. CTB (BeWo) and STB (BeWo+FSK) were treated with or without R-(+)-Methanandamide (0.01µM-10µM Met-AEA), a stable analogous of AEA. Cell viability was evaluated by MTT assay and LDH activity. We studied different trophoblast syncytialization markers: Glial cells missing-1 (GCM-1) mRNA levels by RT-qPCR; human chorionic gonadotropin (hCG) concentration by chemiluminescence immunoassay; syncytin-1 protein expression, cell size and DNA content by flow cytometry; and E-cadherin distribution by immunocytochemistry. Incubation with 1µM Met-AEA diminished the mRNA levels of GCM-1(p