engleski

Amphetamine-like stimulants induce oxidative stress and DNA damage in human neuroblastoma SH-SY5Y cell line

Introduction: Amphetamine-like stimulants (ATS) are widely abused substances that impair dopaminergic and serotonergic functions. The mechanisms of their toxicity in human neuronal cells are not clear yet, but they may involve the formation of reactive oxygen species. The aim of this study was to determine the induction of oxidative stress parameters and the levels of DNA instability in human neuroblastoma SH-SY5Y cells treated with selected concentrations of four different ATS during 24 hours. Methods: SH-SY5Y cells were treated with ATS at concentrations that did not decrease viability below 75 %. A new psychoactive substance mephedrone was tested at 6.25 μM, while so-called classical ATS were applied as follows: 3, 4- methylenedioxymethamphetamine (MDMA, ecstasy) at 0.625 μM, methamphetamine at 0.625 μM, and amphetamine at 3.13 μM. After the treatment, cells were trypsinized, washed in phosphate-buffered saline, resuspended in complete medium and prepared for further analysis. The ATS-toxicity was evaluated using markers of DNA damage and oxidative stress. Lipid peroxidation level (malondialdehyde (MDA)), reactive oxygen species (ROS) production, glutathione (GSH) levels, and activities of antioxidant enzymes (catalase, glutathione peroxidase and superoxide dismutase) were measured as biomarkers of oxidative stress, while primary DNA damage was determined using alkaline comet assay. Results: When compared to controls, cells treated with methamphetamine had a significant increase in MDA, cells treated with mephedrone had a significant increase in ROS, GSH and GPx, whereas cells treated with amphetamine had a significant increase in ROS, and a significant decrease in GSH. The range of DNA damage measured with the comet assay after treatment (based on the median values obtained for tail intensity) was: amphetamine > methamphetamine > mephedrone > MDMA. Conclusions: Significant induction of oxidative stress parameters and increased level of DNA damage detected at cell level call for further studies to clarify the toxicological risks associated with ATS consumption.

amphetamine-like stimulants ; oxidative stress ; DNA damage ; human neuroblastoma SH-SY5Y cell

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