The Cytotoxic Effects of Nyaope, a Heroin-based Street Drug, in SH-SY5Y Neuroblastoma Cells

Abstract

Nyaope is a local adulterated drug that contributes significantly to the psychosocial challenge of substance use in South Africa. Despite being a huge burden on society and the health care system, research into the deleterious effects of nyaope is limited. The aim of the present study was therefore to perform a chemical analysis of the drug and to assess its toxic effects on neuroblastoma cells. Gas chromatography-mass spectrometry (GC/MS) analysis showed that nyaope mainly consists of heroin and heroin-related products. SH-SY5Y cells were subsequently exposed to increasing concentrations of nyaope (0.625, 1.25, 2.5, 5 and 10 µg/µL) for 1, 6 or 24 h. The toxic effects of nyaope were determined by measuring lactate dehydrogenase (LDH) released into the cell culture medium as an indicator of necrosis, the mRNA expression levels of Bax and Bcl-2 as markers of apoptosis, and the mRNA expression levels of p62 and microtubule-associated protein 1 A/1B light-chain 3 (LC3) as indicators of autophagy. Exposing SH-SY5Y cells to concentrations of nyaope 5 µg/µL and greater for 24 h, resulted in a significant increase in LDH levels in the cell culture medium, unchanged mRNA expression of Bax and Bcl-2 mRNA, and significantly reduced p62 and elevated LC3 mRNA expression levels. The chemical analysis suggests that nyaope should be considered synonymous with heroin and the toxic effects of the drug may recruit pathways involved in necrosis and autophagy.

Keywords

• Cell viability
• Necrosis
• Apoptosis
• Autophagy
• Gas chromatography
• PCR