Osmoticc stress and human amnion aquaporins

DOROSSHNKO, BRIAN; DAMIANO, ALICIA E; DI PAOLA, MAURICIO; ANUD, CAROLINA; SIERRA, MATÍAS; MASKIN BERNARDO

Ciudad Autonoma de Buenos Aires

Congreso; IFPA meeting; 2019

International Federation of Placenta Associations

OBJETIVETo study the effect of osmotic stress on the expression and function of human amnion aquaporins (AQPs) and its consequences on cell survival. METHODSThis study was approved by the ethics committee of the Hospital Nacional Prof. Dr. A. Posadas. Human amnion explants and WISH cell line were cultured in hypo (150 mOsm) and hyperosmolar (400 mOsm) conditions. Net transepithelial water movements across the human amnion were measured using a modified Ussing chamber and osmotic permeability (pOsm) was calculated. AQP1, AQP3, AQP8 and AQP9 expressions were assessed by Western Blot and semiquantitative RT-PCR.In some experiments, HgCl2 was used as a general AQPs inhibitor, and Tetraethylammonium (TEA), CuSO4 and phloretin were used as selective inhibitors of AQP1, AQP3 and AQP9, respectively. Cell viability and integrity were studied by the MTT and LDH assays. Nf-kb expression by Western Blot was also assayed.RESULTSIn human amnion, hyperosmolar condition showed a significantly decreased in AQP1 and AQP9 expressions (n=7; p