Avaliação citotóxica de formulações de esteroides anabolizantes apreendidas pela Polícia Federal em cultivo primário de astrócitos

Abstract

Anabolic androgenic steroids (AAAs) are synthetic substances derived from testosterone. Due to the side effects caused by EAAs, their use is controlled or even prohibited. However, their illicit use has further increased the risk of serious deleterious effects. EAAs have neurotoxic actions, however, their effects on astrocytes are still poorly understood. Thus, due to the importance of astrocytes for brain homeostasis, the investigation of the effects of EAAs on these cells becomes relevant. Thus, the objective of this work was to perform the chemical analysis of AAS seized by the Federal Police and to evaluate the cytotoxicity of these compounds in astrocyte culture. Five formulations of EAAs learned by the Federal Police were used (called compounds C1, C2, C3, C4 and C5) and the commercial form of nandrolone decanoate as standard control. The analysis of the active ingredient in the samples was performed using gas chromatography coupled to mass spectrometry (GC-MS). For cytotoxic analysis, cultures of cortical astrocytes from neonatal Wistar rats were exposed to nandrolone ecanoate and to compounds C1 – C5 at concentrations of 10, 30 and 100 μM for 48 and 72 h. Afterwards, cell viability and proliferation tests, and the levels of reactive oxygen species (ROS), total thiol content (SH), nitrite levels and activity of catalase (CAT) and superoxide dismutase (SOD) enzymes were determined. Three samples had a different composition than indicated on the label. C1 and C2 were unlabeled, but nandrolone decanoate was identified in both. In C3, the label indicated the presence of boldenone undecylenate, but nandrolone decanoate was identified in the formulation. At C4, the label indicated Testosterone Propionate, but the compounds identified were Nandrolone Decanoate, Testosterone Propionate, and Testosterone Cypionate. C5 had nandrolone decanoate on the label, but this compound was not identified by GC-MS. Regarding the cytotoxic effects, nandrolone decanoate and C1 increased cell proliferation at a concentration of 10 μM while C3 and C4 at 100 μM (P<0.05). C1 and C2, C4, C5 and nandrolone decanoate (10 μM) and C3 (100 μM) increased cell viability. On the other hand, a reduction in cell viability was observed at the highest concentrations evaluated (P<0.05). An increase in ROS was observed with nandrolone decanoate, C1, C4, C2 and C5 while an increase in nitrites occurred with nandrolone decanoate, C3 and C4 at the highest concentration tested. SH levels were reduced when astrocytes were exposed to nandrolone decanoate, C1, C2 and C5 (P<0.05). Finally, CAT activity was reduced by nandrolone decanoate and C1, C2 and C5 and SOD by C2 and C5 (P<0.05). The apprehended EAAs alter cell viability and proliferation in addition to causing oxidative damage in astrocytes. These findings demonstrate the gliotoxic potential of EAAs in addition to showing that the seized formulations can be dangerous to the health of users, due to their composition being uncertain and not having the correct quality control for their production.