Abstract
Benzodiazepines are highly effective in combating anxiety; however, they have considerable adverse effects, so it is important to discover new safe anxiolytic agents. This study was designed to investigate the effect of the natural product 2-hydroxy-3,4,6-trimethoxyacetophenone (HTMCX) on anxiety and seizure behavior in adult zebrafish and its possible mechanisms of action. The acute toxicity of 96 h of HTMCX was analyzed, and the open and light/dark field tests (n = 6 animals/group) were used to assess the anxiety behavior of animals treated with HTMCX. In addition, the mechanisms of action were investigated with antagonists of the GABAA, 5-HT receptors, and molecular anchorage study. Pentylenetetrazole (PTZ) was used to induce seizure by immersion. As a result, acetophenone HTMCX (1, 3 and 10 mg/kg; v.o.) was non-toxic and affected locomotor activity. The higher doses (3 and 10 mg/kg; v.o.) produced signs of anxiolytic action in the light/dark test, and this effect was reversed by the pizotifen (antagonist 5HTR1 and 5HTR2A/2C), having the potential to form a complex with 5HTR1B. However, the anxiolytic effect of HTMCX has not been abolished by flumazenil (antagonist GABAA), cyproheptadine (antagonist 5HTR2A), and granisetron (antagonist 5HTR3A/3B). Therefore, HTMCX demonstrated an anxiolytic effect, suggesting that the 5HTR1 and 5HTR2C receptors may be involved in the pharmacological performance of this acetophenone in the central nervous system.
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Data availability
All data generated or analyzed during this study are included in the supplementary material.
Code availability
The authors thank Centro Nacional de Processamento de Alto Desempenho (CENAPAD) of the Federal University of Ceará (UFC) for the use of the AutoDock Vina software and to the Centro Nordestino de Aplicação e Uso da Ressonância Magnética Nuclear (CENAUREM) for the NMR measurements. The Discovery Studio Visualizer and UCSF Chimera free software were used to analyzing the results of the research.
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Funding
This research was funded by Fundação Cearense de Apoio ao Desenvolvimento Científico e Tecnológico – FUNCAP (grant number BP4-0172–00075.01.00/20 for Hélcio Silva dos Santos), Coordenação de Aperfeiçoamento de Pessoal de Nível Superior—CAPES (finance code 001 for Antonio Wlisses da Silva), and Conselho Nacional de Desenvolvimento Científico e Tecnológico—CNPq (grant number 305719/2018–1 for Alexandre Magno Rodrigues Teixeira).
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AWS performed the experiments, analyzed the data, prepared figures and tables, and authored and wrote the manuscript. MKAF and JESAM conceived and designed the experiments. ELR, FRSM, and ALSM analyzed the data and reviewed drafts of the manuscript. MMM and ESM performed the docking molecular study and reviewed the paper. HSS and AMRT contributed reagents/materials/analysis tools, supervised the research, and reviewed the manuscript. All authors read and approved the final version of the manuscript. The authors declare that all data were generated in-house and that no paper mill was used.
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All procedures performed in studies involving zebrafish-based animal were approved by the Animal Use Ethics Committee of the State University of Ceará (CEUA-UECE; # 3344801/2017). All methods were performed in accordance with the relevant guidelines and regulations.
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da Silva, A.W., Ferreira, M.K.A., Rebouças, E.L. et al. Anxiolytic-like effect of natural product 2-hydroxy-3,4,6-trimethoxyacetophenone isolated from Croton anisodontus in adult zebrafish via serotonergic neuromodulation involvement of the 5-HT system. Naunyn-Schmiedeberg's Arch Pharmacol 394, 2023–2032 (2021). https://doi.org/10.1007/s00210-021-02116-z
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DOI: https://doi.org/10.1007/s00210-021-02116-z