title: 2-hydroxyoestradiol and 2-methoxyoestradiol, two endogenous oestradiol metabolites, induce DNA fragmentation in Sertoli cells.
abstract: Elevated intratesticular levels of hydroxyoestradiols and methoxyoestradiols, two classes of endogenous oestradiol metabolites, have been associated with male infertility. The aim of this study was to explore the effects of 2-hydroxyoestradiol (2OHE2), 4-hydroxyoestradiol (4OHE2), 2-methoxyoestradiol (2ME2) and 4-methoxyoestradiol (4ME2) on Sertoli cell viability. For this, TM4 cells were incubated with different concentrations of these metabolites for 24 h to then evaluate the viability and DNA integrity by MTS and TUNEL assay respectively. The participation of classical oestrogen receptors and the involvement of oxidative stress and apoptotic mechanisms were also evaluated co-incubating TM4 cells with these estradiol metabolites and with the drugs ICI182780, N-acetylcysteine and Z-VAD-FMK respectively. Only high concentrations of 2OHE2and 2ME2decreased cell viability inducing DNA fragmentation. In addition, ICI182780 did not block the effect of 2OHE2and 2ME2, while N-Acetylcysteine and Z-VAD-FMK only blocked the effect of 2OHE2. Moreover, 2OHE2but not 2ME2induced PARP and caspase-3 cleavage. Finally, lower 2OHE2and 2ME2concentrations (0.01-0.1-1.0 μmol l-1) decreased Sertoli cell viability 48 h post-treatment. Our results support the hypothesis that elevated intratesticular 2OHE2or 2ME2concentrations could be related to male infertility since 2OHE2by apoptosis and 2ME2by undetermined mechanisms induce DNA fragmentation in Sertoli cells.
authors.affiliation: Instituto de Investigaciones Materno Infantil, Facultad de Medicina, Universidad de Chile, Santiago, Chile.
authors.forename: C
authors.initials: C
authors.lastname: Valencia
authors.affiliation: Instituto de Investigaciones Materno Infantil, Facultad de Medicina, Universidad de Chile, Santiago, Chile.
authors.forename: C
authors.initials: C
authors.lastname: Molina
authors.affiliation: Instituto de Investigaciones Materno Infantil, Facultad de Medicina, Universidad de Chile, Santiago, Chile.
authors.forename: M
authors.initials: M
authors.lastname: Florez
authors.affiliation: Departamento de Fisiología, Facultad de Ciencias Biológicas, Pontificia Universidad Católica de Chile, Santiago, Chile.
authors.forename: J
authors.initials: J
authors.lastname: Buñay
authors.affiliation: Departamento de Fisiología, Facultad de Ciencias Biológicas, Pontificia Universidad Católica de Chile, Santiago, Chile.
authors.forename: R D
authors.initials: RD
authors.lastname: Moreno
authors.affiliation: Laboratorio de Inmunología de la Reproducción y CEDENNA, Universidad de Santiago de Chile, Santiago, Chile.
authors.forename: P A
authors.initials: PA
authors.lastname: Orihuela
authors.affiliation: Instituto de Investigaciones Materno Infantil, Facultad de Medicina, Universidad de Chile, Santiago, Chile.
authors.forename: A
authors.initials: A
authors.lastname: Castro
authors.affiliation: Instituto de Investigaciones Materno Infantil, Facultad de Medicina, Universidad de Chile, Santiago, Chile.
authors.forename: A
authors.initials: A
authors.lastname: Parada-Bustamante
mesh_terms: D000077584:2-Methoxyestradiol; D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D053148:Caspase 3 / Q000378:metabolism; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D002648:Child; D053938:DNA Fragmentation / Q000187:drug effects*; D004958:Estradiol / Q000031:analogs & derivatives* / Q000494:pharmacology; D065171:Estrogen Receptor Antagonists / Q000494:pharmacology; D000077267:Fulvestrant; D006801:Humans; D008297:Male; D051379:Mice; D018384:Oxidative Stress / Q000187:drug effects; D000071137:Poly (ADP-Ribose) Polymerase-1 / Q000378:metabolism; D012708:Sertoli Cells / Q000187:drug effects* / Q000378:metabolism
chemical_list: D065171:Estrogen Receptor Antagonists; D000077267:Fulvestrant; D004958:Estradiol; D000077584:2-Methoxyestradiol; C001390:2-hydroxyestradiol; D000071137:Poly (ADP-Ribose) Polymerase-1; D053148:Caspase 3
keywords: Apoptosis; estradiol metabolites; testis