Abstract
N-Ethylhexedrone (NEH) and buphedrone (Buph) are emerging synthetic cathinones (SC) with limited information about their detrimental effects within central nervous system. Objectives: To distinguish mice behavioural changes by NEH and Buph and validate their differential harmful impact on human neurons and microglia. In vivo safety data showed the typical induced behaviour of excitation and stereotypies with 4–64 mg/kg, described for other SC. Buph additionally produced jumping and aggressiveness signs, while NEH caused retropulsion and circling. Transient reduction in body-weight gain was obtained with NEH at 16 mg/kg and induced anxiolytic-like behaviour mainly with Buph. Both drugs generated place preference shift in mice at 4 and 16 mg/kg, suggestive of abuse potential. In addition, mice withdrawn NEH displayed behaviour suggestive of depression, not seen with Buph. When tested at 50–400 μM in human nerve cell lines, NEH and Buph caused neuronal viability loss at 100 μM, but only NEH produced similar results in microglia, indicating different cell susceptibilities. NEH mainly induced microglial late apoptosis/necrosis, while Buph caused early apoptosis. NEH was unique in triggering microglia shorter/thicker branches indicative of cell activation, and more effective in increasing microglial lysosomal biogenesis (100 μM vs. 400 μM Buph), though both produced the same effect on neurons at 400 μM. These findings indicate that NEH and Buph exert neuro-microglia toxicities by distinct mechanisms and highlight NEH as a specific inducer of microglia activation. Buph and NEH showed in vivo/in vitro neurotoxicities but enhanced specific NEH-induced behavioural and neuro-microglia dysfunctionalities pose safety concerns over that of Buph.
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Funding
This work was supported by European Commission-JUST/2014/JDRUG/AG/DRUG and by Fundação para a Ciência e a Tecnologia (FCT): project PTDC-SAU-TOX/32515/2017 and, in part, UID/DTP/04138/2013 (iMed.ULisboa). We also acknowledge the financial support from FCT and Portugal 2020 to the Portuguese Mass Spectrometry Network (LISBOA-01-0145-FEDER-402-022125). ARV was recipient of a Postdoctoral grant from FCT (SFRH/BPD/76590/2011). The funding organizations had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
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AL, DB, CMS and RM were responsible for the study concept and design. CMS, SCH and AL performed the behaviour experiments, acquired and processed animal data. PF performed the synthesis of NPS. ARV, FB, PF and AF performed the cell culture maintenance, incubation with NPS and in vitro data acquisition and processing. CMS, ARV, DB and AL drafted the manuscript. All authors critically reviewed content and approved final version for publication.
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de Mello-Sampayo, C., Vaz, A.R., Henriques, S.C. et al. Designer Cathinones N-Ethylhexedrone and Buphedrone Show Different In Vitro Neurotoxicity and Mice Behaviour Impairment. Neurotox Res 39, 392–412 (2021). https://doi.org/10.1007/s12640-020-00229-6
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DOI: https://doi.org/10.1007/s12640-020-00229-6