BRAIN MICROVASCULAR ENDOTHELIAL RESPONSE TO STIMULI FROM THE PLACENTA. PRELIMINARY RESULTS

LEÓN J.; ESCUDERO C.; REPPETI J.; DAMIANO A.; ACURIO J.; MARTÍNEZ N.

Congreso; III MEETING ON RESEARCH AND INNOVATION IN VASCULAR HEALTH V RESEARCH MEETING ON HYPERTENSION IN PREGNANCY; 2019

Pathophysiology of brain damage in preeclampsia is unclear. But, current hypothesis indicate that hypoxia/reoxygenation induce placental damage that enhances shedding of syncytiotrophoblast microparticles including apoptotic bodies or exosomes into maternal circulation causing brain microvascular endothelium dysfunction. Also, emerging evidence suggests that an intact caveola is necessary for adequate endothelial cell function. Caveola is a small uncoated pit found in the cell membrane of many highly differentiated mammalian cells including endothelium. Within the caveola increasing number of signaling proteins have been documented, including Caveolin-1 (Cav-1) and aquaporin 1 and 4 (AQP1, AQP4) which normal function is related to cell migration and vascular permeability.AimsTo evaluate whether conditioned media from placentas exposed to hypoxia/reoxygenation impairs brain endothelial cells migration and expression of Cav-1, AQP1, AQP4.MethodsHuman brain microvascular cell line hCMEC/D3 was exposed (24 h) to conditioned medium (CM) harvested from normal pregnancy placentas explants cultured in: a) N: normoxia (20% O2); b) H: hypoxia (2% O2); and c) H/R: hypoxia/reoxygenation. Caveolin-1 (Cav-1), aquaporin 1 and 4 (AQP1, AQP4) expressions were evaluated by RT-PCR and Western Blot. To evaluate the role of Cav-1, methyl-β-cyclodextrin (MβCD-5 mM, 30 min) was used to disrupt the caveolae and then hCMEC/D3 migration was evaluated by the wound healing assay (0 to 20 h).ResultsCM from hypoxic placentas significantly increased hCMEC/D3 cell migration at 6h and 8h (n=3; p