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2‐hydroxyoestradiol and 2‐methoxyoestradiol, two endogenous oestradiol metabolites, induce DNA fragmentation in Sertoli cells
Author(s) -
Valencia C.,
Molina C.,
Florez M.,
Buñay J.,
Moreno R. D.,
Orihuela P. A.,
Castro A.,
ParadaBustamante A.
Publication year - 2016
Publication title -
andrologia
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.633
H-Index - 59
eISSN - 1439-0272
pISSN - 0303-4569
DOI - 10.1111/and.12576
Subject(s) - sertoli cell , dna fragmentation , apoptosis , viability assay , andrology , endogeny , tunel assay , chemistry , endocrinology , medicine , spermatogenesis , biology , programmed cell death , biochemistry
Summary 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 (2 OHE 2 ), 4‐hydroxyoestradiol (4 OHE 2 ), 2‐methoxyoestradiol (2 ME 2 ) and 4‐methoxyoestradiol (4 ME 2 ) on Sertoli cell viability. For this, TM 4 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 TM 4 cells with these estradiol metabolites and with the drugs ICI 182780, N‐acetylcysteine and Z‐ VAD ‐ FMK respectively. Only high concentrations of 2 OHE 2 and 2 ME 2 decreased cell viability inducing DNA fragmentation. In addition, ICI 182780 did not block the effect of 2 OHE 2 and 2 ME 2 , while N‐Acetylcysteine and Z‐ VAD ‐ FMK only blocked the effect of 2 OHE 2 . Moreover, 2 OHE 2 but not 2 ME 2 induced PARP and caspase‐3 cleavage. Finally, lower 2 OHE 2 and 2 ME 2 concentrations (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 2 OHE 2 or 2 ME 2 concentrations could be related to male infertility since 2 OHE 2 by apoptosis and 2 ME 2 by undetermined mechanisms induce DNA fragmentation in Sertoli cells.