ANGIOTENSIN II INDUCES DECIDUALISATION MARKERS AND CHEMOATRACTANTS IN HUMAN ENDOMETRIAL STROMAL CELLS AND REGULATES TROPHOBLAST MIGRATION

SOTELO, AGUSTINA; PARRADO, CECILIA; SALAVERRY, LUCIANA; CASTRO, MARISA; REY-ROLDAN, ESTELA; SCOTTO, FLORENCIA; GENTILE, TERESA; CANELLADA, ANDREA

Puerto Varas

Encuentro; Latin American Society for Maternal Fetal Interaction and Placenta (SLIMP), Sociedad Chilena de Ciencias Fisiológicas (SCHCF); 2017

We previously demonstrated that angiotensin II (Ang II) modulates gene expression, proliferation, and metalloprotease activity in rat endometrial stromal cells (ESC). Decidual prolactin and heparin binding-epidermal growth factor (HB-EGF) induced in rat ESC by progesterone-treated trophoblast conditioned medium, decreased in ESC pretreated with losartan.Objective: To analyse a) whether Ang II induces markers of uterine receptivity in human ESC (T-HESC), b) the effect of Ang II treated-T-HESC conditioned medium on the migration of trophoblast HTR8/SVneo cells, c) the involvement of angiotensin receptors.Methods: T-HESC were exposed or not to Ang II (125, 250, 500 ng/ml) for 24, 48, 72, or 96 h. Cells were pretreated (1 h) with INCA-6 (5 μM), as well as with losartan 1 μM or PD123319 (1 μM) to assess the involvement of calcineurin/nuclear factor of activated T cells (CN/NFAT) signalling pathway, and Ang II receptors, respectively. Insulin growth factor binding protein 1 (IGFBP-1) and HB-EGF -mRNA and protein expression was analysed by semiquantitative RT-PCR and Western blot. Interleukin (IL)-1β and IL-8 were analysed in cell culture supernatants by ELISA. HTR8/SVneo trophoblast cells were exposed or not to conditioned medium from T-HESC, which were cultured for 48 h in the presence or absence of Ang II, and their migration on scratch was then registered.Results: Ang II induced the expression of IGFBP-1 respect to control (7.81 ± 2.91 and 7.38 ± 1.52 fold, mRNA and protein, respectively), HB-EGF (1.52 ± 0.12 and 2.86 ± 0.35 fold, mRNA and protein, respectively) and IL-8 (59.29 ± 11.86 vs 23.39 ± 5.45 pg/ml), but not IL-1β in T-HESC. INCA-6 inhibited IGFBP-1 and HB-EGF induction. INCA-6 and losartan, but not PD123319 inhibited the induction of IL-8. Conditioned medium from T-HESC, which was stimulated with Ang II, increased the migration on scratch of HTR8/SVneo. Increased migration was not observed when trophoblast cells were incubated with conditioned medium from T-HESC, which was pretreated with losartan before the Ang II stimuli.Conclusions: Results suggest that Ang II acting at angiotensin receptor-1 induces NFAT-dependent gene expression related with receptivity in human ESC, which could favour the migration of trophoblast cells.