HYPEROSMOLAR STRESS AFFECTS TRPV-1 EXPRESSION AND THE PHYSIOLOGICAL FUNCTIONS OF HUMAN TROPHOBLAST

REPPETTI J.; SEYAHIAN A.; SZPILBARG N.; FARINA M.; DAMIANO A.; MARTÍNEZ N.

Puerto Varas

Simposio; SLIMP- Latin American Society for Maternal Fetal Interaction and Placenta and SCHCF- Sociedad Chilena de Ciencias Fisiológicas; 2017

SLIMP

Hyperosmolar stress may be an important stressor that alters normaldevelopment of embryos or placentation. Transient receptor potentialvanilloid 1 (TRPV-1) is activated by hyperosmolarity and participates inmany cellular processes such as apoptosis, authophagy, and others.Objective: To evaluate the effect of hyperosmolar stress on cell viability,migration and invasion, and the contribution of TRPV-1 to these processesin first trimester human trophoblast cells.Methods: Swan-71 cell line (human trophoblastic cells) was cultured incomplete DMEM-F12 and sucrose hyperosmolar solution was added for24 h with or without capsaicin (CPZ, 1 mM) and capsazepine (CPZ, 10mM). TRPV-1 protein expression was analysed by Western blot. Cellviability was analysed by MTT, LDH and b-hCG assays. Apoptosis wasevaluated by Bax expression, DNA fragmentation and TUNEL assay, andauthophagy by monodansylcadaverine (MDC) assay. Migration wasassessed by wound healing assay, activity of metalloproteinases (MMPs)by zymography, and invasion was evaluated in transwells pre-coatedwith Matrigel.Results: In hyperosmolar conditions, TRPV-1 expression was increased(1.3 ± 0.1 fold) (P<0.05, n ¼ 6), cell viability and b-hCG secretion wasreduced (25 ± 2% and 28 ± 3%, respectively) (P<0.01, n ¼ 8) and LDH levelswere not modified. Apoptotic indices were also increased (1.7 ± 0.1 fold)(P<0.05, n ¼ 6), but no significant changes were observed after theblocking of TRPV-1 with CPZ. MDC assay showed no changes among thetreatments. Finally, cell migration, MMPs activity, and invasion weredecreased (33 ± 4%, 75 ± 8%, and 45 ± 6%, respectively) (P<0.01, n ¼ 6), andthese effects were partially reversed by CPZ.Conclusions: Our results proposed that hyperosmolarity induces cellapoptosis and alters cell migration and invasion processes. Our findingsalso suggest that TRPV-1 may be involved in these events.