The Designer Drug αPHP Affected Cell Proliferation and Triggered Deathly Mechanisms in Murine Neural Stem/Progenitor Cells

SIMPLE SUMMARY: In recent decades, the world has faced an emerging problem, which is assuming increasingly impressive dimensions: the appearance of New Psychoactive Substances, characterized by peculiar pharmacological and toxicological properties, and are exceptionally dangerous for consumers health. Among novel psychoactive substances, synthetic cathinones have recently emerged on the market and, based on their low cost and availability in smartshops and online, their use has been growing extremely, particularly among adolescents and young adults. However, the understanding of neurotoxic mechanisms induced by synthetic cathinones is still lacking, particularly on neural stem/progenitor cell cultures. In the current in vitro study, the effects of increasing αPHP concentrations, one of major MDPV derivatives, on cell viability/proliferation, morphology/ultrastructure, and cell death pathways, have been evaluated after exposure in murine neural stem/progenitor cells, using a battery of complementary techniques. We revealed several different alterations after αPHP treatment, indicating the potential harmful effects of this synthetic cathinone on young brains. Hence, the present investigation could pave the way for a broadened understanding of the synthetic cathinone toxicology needed to establish an appropriate treatment for novel psychoactive substances and the potential consequences for public health. ABSTRACT: Increasing reports of neurological and psychiatric outcomes due to psychostimulant synthetic cathinones (SCs) have recently raised public concern. However, the understanding of neurotoxic mechanisms is still lacking, particularly for the under-investigated αPHP, one of the major MDPV derivatives. In particular, its effects on neural stem/progenitor cell cultures (NSPCs) are still unexplored. Therefore, in the current in vitro study, the effects of increasing αPHP concentrations (25–2000 μM), on cell viability/proliferation, morphology/ultrastructure, genotoxicity and cell death pathways, have been evaluated after exposure in murine NSPCs, using a battery of complementary techniques, i.e., MTT and clonogenic assay, flow cytometry, immunocytochemistry, TEM, and patch clamp. We revealed that αPHP was able to induce a dose-dependent significant decrease of the viability, proliferation and clonal capability of the NSPCs, paralleled by the resting membrane potential depolarization and apoptotic/autophagic/necroptotic pathway activation. Moreover, ultrastructural alterations were clearly observed. Overall, our current findings demonstrate that αPHP, damaging NSPCs and the morpho-functional fundamental units of adult neurogenic niches may affect neurogenesis, possibly triggering long-lasting, irreversible CNS damage. The present investigation could pave the way for a broadened understanding of SCs toxicology, needed to establish an appropriate treatment for NPS and the potential consequences for public health.