Association of electroanalytical and spectrophotometric methods to evaluate the antioxidant activity of isobenzofuranone in primary cultures of hippocampal neurons

Article abstract of DOI:10.1016/j.tiv.2020.104970

The isobenzofuran-1(3H)-ones (phthalides) exhibit various biological activities, including antioxidant activity on reactive oxygen species (ROS). An excess of ROS that cannot be naturally contained by cellular enzymatic systems is called redox imbalance,

Full text of DOI:10.1016/j.tiv.2020.104970

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Association of electroanalytical and spectrophotometric methods
to evaluate the antioxidant activity of isobenzofuranone in
primary cultures of hippocampal neurons
Aniely dos Reis Teixeira, Róbson Ricardo Teixeira, Iara Mariana
Lellis Ribeiro, Wagner Luiz Pereira, Taise Matte Manhabosco,
Ana Carolina Ferreira de Brito, Laser Antônio Machado Oliveira,
Katiane de Oliveira, Pinto Coelho Nogueira
PII:
S0887-2333(20)30520-8
https://doi.org/10.1016/j.tiv.2020.104970
TIV 104970
DOI:
Reference:
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Toxicology in Vitro
Received date:
Revised date:
Accepted date:
25 May 2020
30 July 2020
12 August 2020
Please cite this article as: A. dos Reis Teixeira, R.R. Teixeira, I.M.L. Ribeiro, et al.,
Association of electroanalytical and spectrophotometric methods to evaluate the
antioxidant activity of isobenzofuranone in primary cultures of hippocampal neurons,
Toxicology in Vitro (2020), https://doi.org/10.1016/j.tiv.2020.104970
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Association of electroanalytical and spectrophotometric methods to evaluate the antioxidant
activity of isobenzofuranone in primary cultures of hippocampal neurons
Aniely dos Reis Teixeira1, Róbson Ricardo Teixeira2, Iara Mariana Lellis Ribeiro1, Wagner Luiz
Pereira2, Taise Matte Manhabosco3, Ana Carolina Ferreira de Brito3, Laser Antônio Machado
Oliveira1, Katiane de Oliveira Pinto Coelho Nogueira1
1Neurobiology and Biomaterials Laboratory, Department of Biological Sciences, Federal
University of Ouro Preto, Ouro Preto, MG, Brazil
2 Department of Chemistry, Federal University of de Viçosa, Viçosa, MG, Brazil
3 Department of Physics - REDEMAT, Federal University of Ouro Preto, Ouro Preto, MG, Brazil
*Corresponding author: Katiane Nogueira
E-mail address: katiane@ufop.edu.br
Postal address: Universidade Federal de Ouro Preto, Campus Morro do Cruzeiro, s/n, Instituto de
Ciências Exatas e Biológicas (ICEB), Departamento de Ciências Biológicas (DECBI), Ouro
Preto, Minas Gerais, Brasil. CEP: 35400-000.
Sample CRediT author statement
Katiane Nogueira: Conceptualization, Investigation, Methodology, Validation, Project
administration, Resources, Data Curation, Funding acquisition, Writing – Original Draft, Writing
- Review and Editing, Róbson Teixeira: Conceptualization, Investigation, Methodology,
Validation, Writing – Original Draft, Laser Oliveira: Investigation, Resources, Data Curation,
Taise Manhabosco: Data curation; Formal analysis; Investigation; Methodology; Validation,
Resources Roles/Writing - original draft, Aniely Teixeira: Methodology, Validation, Formal
analysis, Investigation, Data Curation, Writing – Original Draft, Wagner Pereira: Methodology,
Validation, Investigation, Data Curation, Iara Ribeiro: Methodology, Validation, Formal
analysis, Investigation, Ana Carolina Brito: Methodology, Formal analysis, Investigation.

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Abstract
The isobenzofuran-1(3H)-ones (phthalides) exhibit various biological activities, including
antioxidant activity on reactive oxygen species (ROS). An excess of ROS that cannot be
naturally contained by cellular enzymatic systems is called redox imbalance, which damage cell
membranes, proteins, and DNA, thereby possibly triggering neuronal death in several
neurodegenerative diseases. Considering our ongoing efforts to find useful compounds to control
redox imbalance, herein we evaluated the antioxidant activity of two phtalides (compounds 3 and
4), using primary cultures of hippocampal neurons. Spectrophotometric assays showed that
compound 3 significantly reduced (p ≤ 0.05) ROS levels and lipid peroxidation compared to the
control treatment, while compound 4 was unable at any of the tested concentrations. Despite
their structural similarity, these compounds behave differently in the intracellular environment,
which was reliably corroborated by the determination of oxidation potentials via cyclic
voltammetry. It was demonstrated that compound 3 presents a lower oxidation potential. The
combination of the mentioned methods allowed us to find a strong correlation between the
chemical structure of compounds and their biological effects. Taking together, the results
indicate that compound 3 presents desirable characteristics to act as a candidate pharmacological
agent for use in the prevention and treatment of neurodegenerative diseases.
Keywords: spectrophotometric methods, cyclic voltammetry, isobenzofuranones, hippocampal
neurons, free radicals, antioxidants.

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1. Introduction
Primary cell cultures, specifically those of hippocampal neurons, obtained from a region
associated with higher cognitive functions, such as learning and memory, are quite relevant for
studies regarding pathogenic mechanisms involved in neurodegenerative diseases. Unlike cell
lines of the nervous system, in primary cultures, hippocampal neurons become polarized, with
developed axons and dendrites, facilitating the formation of functional synaptic connections
[1,2]. The development of physiological characteristics similar to those observed in vivo results
in these cultures being widely used for studies in the field of neurobiology, including as a model
for studying the neurotoxicity of free radicals as well as the interaction between reactive oxygen
species (ROS) and protective effects of compounds with antioxidant properties [3,4].
Neurotoxicity triggered by redox imbalance is a common feature in the development of different
types of neurodegenerative diseases. Therefore, compounds that can act as preventive treatments
have extensively been tested primarily using spectrophotometric techniques [5,6,7].
Development of new drugs that are capable of curbing redox imbalance and the
consequent damage to cell membranes, proteins, and DNA, without causing neuronal toxicity,
remains a challenge in the field of neurobiology. Considering this context, organic synthesis has
emerged as an alternative that allows structural modifications in compounds that can perform
well in both in vitro and in vivo assays. For instance, in isobenzofuran-1(3H)-ones (phtalides) the
introduction of functional groups at specific molecular sites lead to antiplatelet, cytotoxic,
GABA receptor-antagonistic, antimicrobial, antifungal, or antioxidant activity [8], demonstrating
that structural similarities between compounds do not necessarily indicate effectiveness or
similar function.