Primary hepatocytes as an useful bioassay to characterize metabolism and bioactivity of illicit steroids in cattle
TOXICOLOGY IN VITRO
Authors: Giantin, Mery; Gallina, Guglielmo; Pegolo, Sara; Lopparelli, Rosa Maria; Sandron, Clara; Zancanella, Vanessa; Nebbia, Carlo; Favretto, Donata; Capolongo, Francesca; Montesissa, Clara; Dacasto, Mauro
Abstract
Cattle hepatocytes have already been used in veterinary in vitro toxicology, but their usefulness as a multi-parametric screening bioassay has never been investigated so far. In this study, cattle hepatocytes were incubated with illicit steroids/prohormones (boldenone, BOLD; its precursor boldione, ADD; dehydroepiandrosterone, DHEA; an association of ADD:BOLD), to characterize their transcriptional effects on drug metabolizing enzymes (DMEs) and related nuclear receptors (NRs), on cytochrome P450 3A (CYP3A) apoprotein and catalytic activity as well as to determine ADD and BOLD metabolite profiling. DHEA-exposed cells showed an up-regulation (higher than 2.5-fold changes) of three out of six NRs, CYP2B22 and CYP2C87; likewise, ADD:BOLD increased CYP4A11 mRNA levels. In contrast, a reduction of CYP1A1 and CYP2E1 mRNAs (lower than 2.5(-1)-fold changes) was noticed in ADD- and DHEA-incubated cells. No effect was noticed on CYP3A gene and protein expression, though an inhibition of 6 beta-, 2 beta- and 16 beta-hydroxylation of testosterone (higher than 60% of control cells) was observed in ADD- and BOLD-exposed cells. Finally, 17 alpha-BOLD was the main metabolite extracted from hepatocyte media incubated with ADD and BOLD, but several mono-hydroxylated BOLD and ADD derivatives were detected, too. Collectively, cattle hepatocytes can represent a complementary screening bioassay, useful to characterize growth promoters metabolite profiling and their effects upon DMEs expression, regulation and function. (C) 2012 Elsevier Ltd. All rights reserved.
Impacts of dose and time of boldenone and stanazolol exposure in inflammatory markers, oxidative and nitrosative stress and histopathological changes in the rat testes
THERIOGENOLOGY
Authors: Bueno, Andressa; Carvalho, Fabiano B.; Gutierres, Jessie M.; Lhamas, Cibele L.; Brusco, Indiara; Oliveira, Sara M.; Amaral, Marta G.; Dorneles, Guilherme; Sorraila, Juliana; Duarte, Marta M.; de Andrade, Cinthia M.
Abstract
The present study was conducted to analyze the adverse effects of the anabolic steroids boldenone (BOL) and stanazolol (ST) in the reproductive function of male rats. These molecules were administered using three different protocols. In Protocol I, BOL and ST were administered in a higher dose than what is recommended but for a short period. In Protocol II, a moderate dose of these compounds was applied for an intermediate period, whereas in Protocol III a reduced dose was administered but for an extended period. Notably, Protocol I and III resulted in increased levels of reactive oxygen specimens (ROS [I, p < 0.01] [III, p < 0.001)]) and nitrite plus nitrate (NOx [I, p < 0.01] [II, p < 0.01] [III, p < 0.05]), respectively, whereas non-protein thiols (NPSH) levels were decreased only after Protocol III (p < 0.01). Myeloperoxidase activity was significantly increased after treatment with BOL in protocol II (p < 0.01) and III (p < 0.05) than with ST in protocol III (p < 0.05). Boldenone and ST also caused a significant up-regulation in the levels of serum testosterone when protocols I (p < 0.01) and II (p < 0.05) were performed. There were also visible histopathological alterations in the testes induced by treatment with BOL, namely degenerative changes primarily characterized by a decrease in the germinal epithelium. Together, these results suggest that the administration of BOL or ST exerts a significantly harmful effect in the testes of male rats. Moreover, all the treatment protocols used in this study induced deleterious effects on the testes, as indicated by the different biochemical parameters investigated. However, only the protocols of longer exposure time (II and III) induced morphological changes compatible with infertility. (c) 2017 Elsevier Inc. All rights reserved.