Abstract
Chlorpyrifos (CPF) is a neurotoxic organophosphorus (OP) insecticide widely used for agricultural purposes. CPF-mediated neurotoxicity is mainly associated with its anticholinesterase activity, which may lead to a cholinergic syndrome. CPF metabolism generates chlorpyrifos-oxon (CPF-O), which possesses higher anticholinesterase activity and, consequently, plays a major role in the cholinergic syndrome observed after CPF poisoning. Recent lines of evidence have also reported non-cholinergic endpoints of CPF- and CPF-O-induced neurotoxicities, but comparisons on the non-cholinergic toxic properties of CPF and CPF-O are lacking. In this study, we compared the non-cholinergic toxicities displayed by CPF and CPF-O in cultured neuronal cells, with a particular emphasis on their pro-oxidant properties. Using immortalized cells derived from mouse hippocampus (HT22 line, which does present detectable acetylcholinesterase activity), we observed that CPF-O was 5-fold more potent in decreasing cell viability compared with CPF. Atropine, a muscarinic acetylcholine receptor antagonist, protected against acetylcholine (ACh)-induced toxicity but failed to prevent the CPF- and CPF-O-induced cytotoxicities in HT22 cells. CPF or CPF-O exposures significantly decreased the levels of the antioxidant glutathione (GSH); this event preceded the significant decrease in cell viability. Pretreatment with N-acetylcysteine (NAC, a GSH precursor) protected against the cytotoxicity induced by both CPF and CPF-O. The present study indicates that GSH depletion is a non-cholinergic event involved in CPF and CPF-O toxicities. The study also shows that in addition of being a more potent AChE inhibitor, CPF-O is also a more potent pro-oxidant molecule when compared with CPF, highlighting the role of CPF metabolism (bioactivation to CPF-O) in the ensuing non-cholinergic toxicity.
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Acknowledgements
The project CNPq-PVE (300966/2014-8), afforded by CNPq and CAPES, is specially acknowledged. We are grateful to the Laboratório Multiusuário de Estudos em Biologia at the Universidade Federal de Santa Catarina (LAMEB/UFSC) for providing its infrastructure for carrying out biochemical analysis.
Funding
This work was supported by grants from the (i) Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq), (ii) Fundação de Apoio à Pesquisa do Estado de Santa Catarina, and (iii) Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES). M.W.L. was a PNPD/CAPES postdoctoral grantee. Scholarships to A.A.N and M.W.L (CAPES) are also acknowledged. ALD, JBTR, RBL, and MF receive CNPq-research scholarships.
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Highlights
• Chlorpyrifos and chlorpyrifos-oxon caused toxicity in HT22 cells.
• Chlorpyrifos-oxon is more toxic to HT22 cells when compared with chlorpyrifos.
• Atropine does not prevent such toxicity.
• Chlorpyrifos and chlorpyrifos-oxon caused glutathione depletion.
• N-acetylcysteine prevented chlorpyrifos- and chlorpyrifos-oxon-induced toxicity.
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Naime, A.A., Lopes, M.W., Colle, D. et al. Glutathione in Chlorpyrifos-and Chlorpyrifos-Oxon-Induced Toxicity: a Comparative Study Focused on Non-cholinergic Toxicity in HT22 Cells. Neurotox Res 38, 603–610 (2020). https://doi.org/10.1007/s12640-020-00254-5
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DOI: https://doi.org/10.1007/s12640-020-00254-5