Effects of Bisphenol A, Benzophenone 2 or Benzophenone 3 on Autophagy Protein Markers in Immortalized Gonadotropin-Releasing Hormone (GnRH) Neurons

JUAN MANUEL RIAÑO GÓMEZ; VICTORIA AR LUX-LANTOS; ELEONORA M. SORIANELLO; MARINA O. FERNANDEZ

Mar del Plata

Congreso; REUNIÓN ANUAL DE SOCIEDADES DE BIOCIENCIAS 2023; 2023

Benzophenones (BP) and Bisphenol A (BPA) are endocrine disrupting chemicals (EDC). BPA is a monomer of polycarbonate plastics and BPs are UV filters. Previously, we showed that the in vitro exposure to EDC inhibited kisspeptin-10-induced GnRH gene expression, and altered expression of inflammatory markers in GT1-7 cells; it also modified gene expression of autophagy markers in GnRH-expressing cell lines. In this study, we evaluated the effects of the exposure to BPA, BP2 or BP3 (1x10-7 or 1x10-9 M) on protein expression of autophagy markers, LC3 and p62.GnRH-expressing cell lines, GT1-7 or GN11 cells, were plated in 6-well plates, in DMEM high glucose, with 10% FBS and antibiotics. Twenty-four hours later, media was replaced by phenol red-free DMEM with 10% charcoal-stripped FBS and antibiotics, and cells were exposed to BPA, BP2 or BP3 (1x10-7 or 1x10-9 M), alone or in combination with Chloroquine (CQ, an inhibitor of the degradation of the autophagosome content), for 12 or 24h. Media were removed and plates were frozen at -70 C for western blot analysis. Cells were lysed in RIPA buffer with protease inhibitors and protein concentration measured by Bradford. Proteins were separated in polyacrylamide gels and transferred to PVDF membranes. LC3, p62, and tubulin were detected with specific antibodies. Results were expressed as Media±SEM and analyzed by Repeated Measures Two-way ANOVA or by ANOVA (Statistica).In GT1-7 cells, CQ increased the lipidated form of LC3 (LC3-II, p