title: In vitro effect of 4-nonylphenol on human chorionic gonadotropin (hCG) stimulated hormone secretion, cell viability and reactive oxygen species generation in mice Leydig cells.
abstract: Nonylphenol is considered an endocrine disruptor and has been reported to affect male reproductive functions. In our in vitro study, we evaluated the effects of 4-nonylphenol (4-NP) on cholesterol levels, hormone formation and viability in cultured Leydig cells from adult ICR male mice. We also determined the potential impact of 4-NP on generation of reactive oxygen species (ROS) after 44 h of cultivation. The cells were cultured with addition of 0.04; 0.2; 1.0; 2.5 and 5.0 μg/mL of 4-NP in the present of 1 IU/mL human chorionic gonadotropin (hCG) and compared to the control. The quantity of cholesterol was determined from culture medium using photometry. Determination of hormone production was performed by enzyme-linked immunosorbent assay. Metabolic activity assay was used for quantification of cell viability. The chemiluminescence technique, which uses a luminometer to measure reactive oxygen species, was employed. Applied doses of 4-NP (0.04-5.0 μg/mL) slight increase cholesterol levels and decrease production of dehydroepiandrosterone after 44 h of cultivation, but not significantly. Incubation of 4-NP treated cells with hCG significantly (P < 0.001) inhibited androstenedione, but not testosterone, formation at the highest concentration (5.0 μg/mL). The viability was significantly (P < 0.05); (P < 0.001) increased at 1.0; 2.5 and 5.0 μg/mL of 4-NP after 44 h treatment. Furthermore, 44 h treatment of 4-NP (0.04-5.0 μg/mL) caused significant (P < 0.001) intracellular accumulation of ROS in exposed cells. Taken together, the results of our in vitro study reported herein is consistent with the conclusion that 4-nonylphenol is able to influence hormonal profile, cell viability and generate ROS.
authors.affiliation: Department of Animal Physiology, Faculty of Biotechnology and Food Sciences, Slovak University of Agriculture in Nitra, Tr. A. Hlinku 2,94976 Nitra, Slovakia Republic. Electronic address: tomasjambor1@gmail.com.
authors.forename: Tomáš
authors.initials: T
authors.lastname: Jambor
authors.affiliation: Department of Animal Physiology, Faculty of Biotechnology and Food Sciences, Slovak University of Agriculture in Nitra, Tr. A. Hlinku 2,94976 Nitra, Slovakia Republic. Electronic address: evina.tvrda@gmail.com.
authors.forename: Eva
authors.initials: E
authors.lastname: Tvrdá
authors.affiliation: Department of Animal Physiology, Faculty of Biotechnology and Food Sciences, Slovak University of Agriculture in Nitra, Tr. A. Hlinku 2,94976 Nitra, Slovakia Republic. Electronic address: tusimova.eva@gmail.com.
authors.forename: Eva
authors.initials: E
authors.lastname: Tušimová
authors.affiliation: Department of Animal Physiology, Faculty of Biotechnology and Food Sciences, Slovak University of Agriculture in Nitra, Tr. A. Hlinku 2,94976 Nitra, Slovakia Republic. Electronic address: anton.kovacik@yahoo.com.
authors.forename: Anton
authors.initials: A
authors.lastname: Kováčik
authors.affiliation: Department of Animal Physiology, Faculty of Biotechnology and Food Sciences, Slovak University of Agriculture in Nitra, Tr. A. Hlinku 2,94976 Nitra, Slovakia Republic. Electronic address: jana312@gmail.com.
authors.forename: Jana
authors.initials: J
authors.lastname: Bistáková
authors.affiliation: National Institute of Chemical Safety, Nagyvaradter 2, H-1450, Budapest, Hungary. Electronic address: zforgacs@gmail.com.
authors.forename: Zsolt
authors.initials: Z
authors.lastname: Forgács
authors.affiliation: Department of Animal Physiology, Faculty of Biotechnology and Food Sciences, Slovak University of Agriculture in Nitra, Tr. A. Hlinku 2,94976 Nitra, Slovakia Republic. Electronic address: norbert.lukac@uniag.sk.
authors.forename: Norbert
authors.initials: N
authors.lastname: Lukáč
mesh_terms: D000735:Androstenedione / Q000378:metabolism; D000818:Animals; D002470:Cell Survival / Q000187:drug effects*; D006063:Chorionic Gonadotropin / Q000378:metabolism*; D052244:Endocrine Disruptors / Q000494:pharmacology*; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D007985:Leydig Cells / Q000187:drug effects* / Q000502:physiology; D008297:Male; D051379:Mice; D008813:Mice, Inbred ICR; D010636:Phenols / Q000378:metabolism / Q000494:pharmacology*; D017382:Reactive Oxygen Species / Q000378:metabolism; D013739:Testosterone / Q000378:metabolism
chemical_list: D006063:Chorionic Gonadotropin; D052244:Endocrine Disruptors; D010636:Phenols; D017382:Reactive Oxygen Species; D013739:Testosterone; D000735:Androstenedione; C041594:4-nonylphenol
keywords: 4-nonylphenol; Hormones; Human chorionic gonadotropin; Reactive oxygen species; Viability