In vitro assessment of the cytotoxic, genotoxic and oxidative stress effects of the synthetic cannabinoid JWH-018 in human SH-SY5Y neuronal cells

Abstract

Background: JWH-018 was the first synthetic cannabinoid introduced as a legal high and the first of the new generation of novel psychoactive substances that flooded worldwide drug markets. JWH-018 was marketed as “spice,” “herbal incense,” or “herbal blend,” as a popular and legal (at the time) alternative to cannabis (marijuana). JWH-018 is a potent synthetic cannabinoid with considerable toxicity associated with its use. JWH-018 has qualitatively similar but quantitatively greater pharmacological effects than cannabis, leading to intoxications and even deaths. The mechanisms of action of the drug’s toxicity require research, and thus, the aim of the present study was to investigate the toxicological profile of JWH-018 in human SH-SY5Y neuronal cells. Methods: SH-SY5Y neuronal cells were exposed to increasing concentrations from 5 to 150 μM JWH-018 over 24 h. Cytotoxicity, DNA damage, the apoptotic/necrotic rate, and oxidative stress were assessed following SH-SY5Y exposure. Results: JWH-018 did not produce a significant decrease in SH-SY5Y cell viability, did not alter apoptotic/necrotic rate, and did not cause genotoxicity in SH-SY5Y cells with 24-h exposure. Glutathione reductase and catalase activities were significantly reduced; however, there was no significant change in glutathione peroxidase activity. Also, JWH-018 treatment significantly decreased glutathione concentrations, significantly increased protein carbonylation, and significantly increased malondialdehyde (MDA) concentrations. For significance, all P < 0.05. Discussion/Conclusion: JWH-018 produced oxidative stress in SH-SY5Y cells that could be an underlying mechanism of JWH-018 neurotoxicity. Additional in vivo animal and human-based studies are needed to confirm our findings.

Human Neuronal Cell Lines as An In Vitro Toxicological Tool for the Evaluation of Novel Psychoactive Substances

“…The treatment with fentanyl induces a high number of either annexin V-FITC and/or PI-stained cells, and increased levels of Bax and caspase 3 activity, supporting a double mechanism of cell death. It should be noted that the concentrations of drugs used in our in vitro study are higher than those found in urine or blood of drug abusers [85][86][87]; however, our data are consistent with several studies reporting in vitro cytotoxic effects (i.e., reduced viability at concentrations >1-2 mM) after exposure to several drugs of abuse, including cathinones, amphetamine-like stimulants, phenethylamines, and synthetic cannabinoids [41,[88][89][90][91][92]. It should also be considered that, due to their high lipophilicity, fentanyl and cathinone derivatives possess a high volume of distribution, reaching significantly greater concentrations in lipophilic tissues, such as the central nervous system [93], and consequently, their blood concentrations might not directly reflect their organ/tissue levels.…”

The Anticancer Effect of Cannabinoid 2 Agonist L-759,633 on C6 and SH-SY5Y Cell Lines

“…This is in contrast to the study conducted by Wojcieszak et al (2016) who demonstrated that CB2 receptor agonist JWH-133 produced a concentration-dependent decline of SH-SY5Y cell viability and reproduction rate, and in contrast to the study conducted by Fisher et al (2016) who showed that medication with cannabidiol (CBD) reduced the viability and invasiveness of NBL cells and stimulated apoptosis in vitro. In line with our study, Sezer et al (2021) indicated that JWH-018, a non selective cannabinoid agonist, did not cause a significant reduction, in SH-SY5Y cell viability, did not modify apoptotic/necrotic rate, and did not induce genotoxicity in SH-SY5Y cells with one day exposure; Cioni et al (2019) demonstrated that COR167 significantly decreased the proliferation of both anaplastic astrocytoma and glioblastoma in a dose-dependent manner; Sánchez et al (2001) indicated that JWH-133 decreased the growth of tumors derived from C6 cell line, but this effect was prevented by the selective CB2 antagonist SR144528. The level of reactive oxygen species is one of the factors that play an significant role in the development and metastasis of tumors.…”

Novel Psychoactive Substances: The Razor’s Edge between Therapeutical Potential and Psychoactive Recreational Misuse

“…Despite their potential therapeutic use in a wide variety of pathologies and conditions, synthetic cannabinoids are associated with toxicity and several adverse effects [ 266 , 267 , 268 , 269 , 270 , 271 , 272 , 273 , 274 , 275 ]. Focusing on the synthetic cannabinoids that demonstrated medical potential, namely those belonging to the JWH, HU and AM series, the present section describes the toxicological effects observed.…”

“…Moreover, the cellular mechanism underlying this effect was explored. Sezer and collaborators observed an increment in oxidative stress by increasing MDA levels, decreasing the activity of glutathione reductase (GR) and catalase, responsible for the detoxification of peroxide hydrogen (H 2 O 2 ), reducing glutathione levels (GSH), a known endogenous antioxidant that protect cells from oxidative reactions [ 269 ]. Based on these results and considering that studies indicate that JWH-133 has therapeutic potential, the authors call for caution in its possible use as this substance may accumulate in the CNS and lead to neuronal damage [ 266 ].…”

“…Based on these results and considering that studies indicate that JWH-133 has therapeutic potential, the authors call for caution in its possible use as this substance may accumulate in the CNS and lead to neuronal damage [ 266 ]. Contrastingly, in the same cellular model, Couceiro et al and Sezer et al found that synthetic cannabinoid JWH-018 does not significantly modify the cell viability of SH-SY5Y cells [ 269 , 270 ]. Later, using yeast as a model, it was observed that this synthetic cannabinoid increases the rate of cell proliferation significantly through an increase in glycolytic flow to the detriment of the pentose phosphate pathway, which suffers a decrease [ 271 ].…”