High Levels of Tumor Necrosis Factor-Alpha Reduces Placental Aquaporin 3 Expression and Impairs in vitro Trophoblastic Cell Migration

RODRIGUES DOS PASSOS, R.; ALVES DE FREITAS, R.; REPPETTI J.; MEDINA Y.; DELA JUSTINA, V.; BACH C.; BOMFIM G.; VITORINO LIMA V.; DAMIANO A. E.; GIACHINI F.R.

Simposio; International Symposium on Reproductive Health: overcoming barriers for research in reproduction; 2021

High Levels of Tumor Necrosis Factor-Alpha Reduces Placental Aquaporin 3 Expression and Impairs in vitro Trophoblastic Cell MigrationRinaldo R. dos Passos Junior1, Raiany A. de Freitas1, Julieta Reppetti 3, Yollyseth Medina3, Vanessa Dela Justina1, Camila W. Bach2, Gisele F. Bomfim5, Victor V. Lima2, Alicia E. Damiano3,4, Fernanda R. Giachini 1,21 ? Institute of Biological Sciences, Federal University of Goias, Goiânia, Goiás, Brazil.2 ? Institute of Biological Sciences and Health, Federal University of Mato Grosso, Barra do Garças, Mato Grosso, Brazil.3 ? Institute of Physiology and Biophysics Bernardo Houssay (IFIBIO) ? CONICET, Faculty of Medicine, University of Buenos Aires, Buenos Aires, Argentina4 ? Department of Biological Sciences, Faculty of Pharmacy and Biochemistry, University of Buenos Aires, Buenos Aires, Argentina.5 ? Institute of Health Sciences, Federal University of Mato Grosso, Sinop, Mato Grosso, Brazil.Introduction: Placentas from preeclamptic women display augmented tumor necrosis factor-alpha (TNF-α) levels with reduced expression of aquaporin 3 (AQP3). However, whether TNF-α modulates AQP3 expression remains to be elucidated. Objective: To characterize if elevated levels of TNF-α reduces AQP3 expression and negatively impacts on trophoblastic cell migration. Methods: Spontaneously hypertensive rats (SHR) and Wistar (14-16 wk) were divided into hypertensive (n=6) and normotensive (n=6) groups. Systolic blood pressure (SBP) was measured, and animals mated. In a third group, pregnant SHRs (n=8) were treated with a TNF-α antagonist, etanercept (0.8 mg/kg, i.p.) on days 0, 6, 12, and 18 of pregnancy. Placentas were collected on the 20th day of pregnancy. Human placental explants (n=7), from normotensive pregnancies, were incubated with TNF-α (5, 10 and 20 ng/mL) and/or etanercept (1 µg/mL). Swan 71 cells were incubated with TNF-α (10 ng/mL) and/or etanercept (1 µg/mL) and subjected to the wound healing assay. AQP3 expression was assessed by western blot and TNF-α levels by ELISA. Results: SBP (mmHg) was elevated in the hypertensive group, and etanercept treatment reduced this parameter. Placental TNF-α levels (pg/mL) were higher in the hypertensive group. AQP3 expression was reduced in the hypertensive group, and etanercept treatment reversed this parameter. Explants submitted to TNF-α exposition displayed reduced expression of AQP3 and etanercept incubation reversed it. Trophoblastic cells incubated with TNF-α showed decreased cell migration and reduced AQP3 expression and etanercept incubation ameliorated it. Conclusion: Altogether, these data demonstrate that high TNF-α levels negatively modulates AQP3 in placental tissue, impairing cell migration, and its relationship in a pregnancy affected by hypertension. Key Words: placenta, hypertensive pregnancy, preeclampsia, TNF-α, trophoblast migration.