Couceiro Joana, Bandarra Susana, Sultan Haider, Bell Suzanne, Constantino Susana, Quintas Alexandre
Centro de Investigação Interdisciplinar Egas Moniz, Instituto Superior de Ciências da Saúde Egas Moniz, 2825-084 Caparica, Portugal; Laboratório de Ciências Forenses e Psicológicas Egas Moniz, Campus Universitário-Quinta da Granja, Monte de Caparica, 2825-084 Caparica, Portugal.
Centro de Investigação Interdisciplinar Egas Moniz, Instituto Superior de Ciências da Saúde Egas Moniz, 2825-084 Caparica, Portugal.
Forensic Sci Int. 2016 Jul;264:100-5. doi: 10.1016/j.forsciint.2016.03.024. Epub 2016 Mar 18.
The emergence and abuse of synthetic cannabinoids has been increasing as an alternative to cannabis, mainly among youth. As their appearance on the drug market has been recent, the pharmacological and toxicological profiles of these psychoactive substances are poorly understood. Current studies suggest that they have stronger effects compared to their natural alternatives and their metabolites retain affinity towards CB1 receptors in CNS. Since studies on its toxicological properties are scarce, the effects of the drug in human derived cell lines were investigated. The present study was designed to explore the toxicological impact of parent drug versus phase I metabolites of synthetic cannabinoids on human cells with and without CB1 receptor. The human cell line of neuroblastoma SH-SY5Y and human kidney cell line HEK-293T were exposed to JWH-018 and to its N-(3-hydroxypentyl) metabolite. Cell toxicity was evaluated using the MTT and LDH assay. Additionally, a dual staining methodology with fluorescent Annexin V-FITC and propidium iodide was performed to address the question of whether JWH-018 N-(3-hydroxypentyl) metabolite is inducing cell death through apoptosis or necrosis, in HEK293T and SH-SY5Y cell lines. The obtained results show that JWH-018 does not cause a statistically significant decrease in cell viability, in contrast to its N-(3-hydroxypentyl) metabolite, which at ≥25μM causes a significant decrease in cell viability. Both cell lines are affected by JWH-018 metabolite. Our results point to higher toxicity of JWH-018 metabolite when compared to its parent drug, suggesting a non-CB1 receptor mediated toxicological mechanism. Comparing the results from Annexin V/PI with MTT and LDH assays of SH-SY5Y and HEK293T in the presence of the synthetic cannabinoid metabolite, emerges the picture that cellular viability decreases and associated death is occurring through necrosis.
作为大麻的替代品，合成大麻素的出现和滥用情况一直在增加，主要发生在年轻人当中。由于它们最近才出现在毒品市场上，人们对这些精神活性物质的药理和毒理学特性了解甚少。目前的研究表明，与天然替代品相比，它们的作用更强，并且其代谢产物对中枢神经系统中的CB1受体仍具有亲和力。由于关于其毒理学特性的研究很少，因此对该药物在人源细胞系中的作用进行了研究。本研究旨在探讨合成大麻素的母体药物与其I期代谢产物对有或无CB1受体的人类细胞的毒理学影响。将神经母细胞瘤SH-SY5Y人细胞系和人肾细胞系HEK-293T暴露于JWH-018及其N-(3-羟基戊基)代谢产物中。使用MTT和LDH试验评估细胞毒性。此外，还采用了荧光膜联蛋白V-FITC和碘化丙啶的双重染色方法，以研究JWH-018的N-(3-羟基戊基)代谢产物是否通过凋亡或坏死诱导HEK293T和SH-SY5Y细胞系中的细胞死亡。获得的结果表明，与N-(3-羟基戊基)代谢产物相比，JWH-018不会导致细胞活力出现统计学上的显著下降，而N-(3-羟基戊基)代谢产物在浓度≥25μM时会导致细胞活力显著下降。两种细胞系均受到JWH-018代谢产物的影响。我们的结果表明，JWH-018代谢产物与其母体药物相比具有更高的毒性，提示存在一种非CB1受体介导的毒理学机制。在存在合成大麻素代谢产物的情况下，将SH-SY5Y和HEK293T的膜联蛋白V/PI结果与MTT和LDH试验结果进行比较，结果显示细胞活力下降，且相关死亡是通过坏死发生的。
