Ascorbic Acid Prevents Efavirenz-Induced Anxiety-Like Behavior and Brain Oxidative Stress in Zebrafish

Abstract

Efavirenz (EFV) is a medication widely used for the treatment of HIV-positive patients. Several studies have demonstrated that the prolongate use of EFV can lead to the development of neurological diseases, such as panic syndrome, depression, and anxiety disorders. In this current study, we evaluate whether the ascorbic acid (AA) treatment can prevent anxiety-like behavior and brain oxidative stress induced by EFV treatment in zebrafish. Our data demonstrated that the EFV treatment induces anxiogenic-like behavior and intense lipid peroxidation in the zebrafish brain. The AA treatment was able to prevent both anxiogenic-like behavior and brain oxidative stress elicited by the EFV treatment. Therefore, our data provide robust evidence that the EFV induced anxiety-like behavior in zebrafish via a redox-dependent pathway and that AA treatment can minimize these adverse effects. Taken together, our preclinical study strongly suggests that the use of an AA-enriched diet can minimize the effects of EFV on the central nervous system (CNS) and improve the quality of life for patients undergoing EFV treatment.

Keywords: anxiety-like behavior; ascorbic acid; efavirez; oxidative stress; zebrafish.

Figure 1

Timeline of the experimental procedures, starting with acclimatization, pharmacological applications followed by behavioral tests, and brain collection for the biochemical assays. (Illustration produced using BioRender.com).

Figure 2

Effect of Efavirenz (EFV) treatment on zebrafish. (a) Locomotion in the dark and white side of aquarium, (b) time spent in the white compartment, (c) freezing, (d) total distance, (e) thigmotaxis, and (f) mean speed. Control saline solution (CTRL) 0.9% (n = 14) and EFV: 30 mg/kg (n = 14). Graphs represented as mean ± SEM and comparisons were made using Student's t-test. ⁣^∗∗∗∗p  < 0, 00001 vs. CTRL.

Figure 3

Lipid peroxidation in the zebrafish brain. Malondialdehyde levels in brain tissue of (a) control and EFV group and (b) co-treated and pretreated with AA + EFV. Control saline solution (CTRL) 0.9% (n = 14), AA: 2 mg/kg (n = 12), EFV: 30 mg/kg (n = 10), co-AA + EFV (n = 10), and pre-AA + EFV (n = 9). Data are expressed as a percentage of control ± SEM. For comparison between two groups, the Student's t-test was used and for more than two groups, one-way ANOVA was performed, followed by Tukey post hoc test. ⁣^∗∗∗∗p  < 0.00001, ⁣^∗∗∗p  < 0.0001, and ⁣^∗p ≤ 0.05.

Figure 4

Effect of AA on zebrafish treated with EFV. (a) Locomotion in the dark and white side of aquarium, (b) time spent in the white compartment, (c) freezing, (d) total distance, (e) thigmotaxis, and (f) mean speed. Control saline solution (CTRL) 0.9% (n = 14), AA: 2 mg/kg (n = 12), EFV: 30 mg/kg (n = 14), co-AA + EFV (n = 12), and pre-AA + EFV (n = 9). The comparison between three or more groups was made by analysis of variance (ANOVA) followed by Bonferroni post hoc test. Graphs represented as mean ± SEM. ⁣^∗∗∗∗p  < 0, 00001, ⁣^∗∗∗p  < 0, 0001, and ⁣^∗∗p  < 0, 001.