Behavioral and Molecular Analysis of Antioxidative Potential of Rosmarinic Acid Against Methamphetamine-induced Augmentation of Casp3a mRNA in the Zebrafish Brain

Vida Nikshenas Shahrestani; Mohammad Haddadi; Ali Reza Samzadeh Kermani

doi:10.32598/bcn.12.2.1777.1

Behavioral and Molecular Analysis of Antioxidative Potential of Rosmarinic Acid Against Methamphetamine-induced Augmentation of Casp3a mRNA in the Zebrafish Brain

Basic Clin Neurosci. 2021 Mar-Apr;12(2):243-254. doi: 10.32598/bcn.12.2.1777.1. Epub 2021 Mar 1.

Authors

Vida Nikshenas Shahrestani¹, Mohammad Haddadi¹, Ali Reza Samzadeh Kermani²

Affiliations

¹ Department of Biology, Faculty of Basic Sciences, University of Zabol, Zabol, Iran.
² Department of Chemistry, Faculty of Basic Sciences, University of Zabol, Zabol, Iran.

Abstract

Introduction: Methamphetamine (MA) acts as a powerful oxidant agent, while Rosmarinic Acid (RA) is an effective herbal antioxidant. Oxidative stress-mediated by MA results in apoptosis, and caspase-3 is one of the critical enzymes in the apoptosis process. MA can epigenetically alter gene regulation. In this paper, to investigate the effects of RA on MA-mediated oxidative stress, changes in the level of casp3a mRNA were demonstrated in zebrafish.

Methods: The animals were grouped in 3 treatment conditions for the behavioral test: control, MA, MA pretreated by RA, and 6 treatment conditions for the molecular test: control, RA, MA, MA co-treated with RA, MA co-treated with RA/ZnO/chitosan nanoparticle, and ZnO/chitosan nanoparticle. Then molecular and behavioral investigations were carried out, and critical comparisons were made between the groups.MA solution was prepared with a concentration of 25 mg/L, and RA solution was prepared by DPPH test with the antioxidant power of about 97%. Each solution was administered by immersing 20 zebrafish for 20 minutes, once per day for 7 days. The level of casp3a mRNA was quantified by using qRT-PCR. One-sided trapezoidal tank diving test was applied to study behavioral alterations.

Results: The qPCR analysis demonstrated the high potential of RA/ZnO/chitosan in counteracting the MA-mediated elevation in casp3a mRNA level. Based on the diving test results of MA-treated fish, MA was found to be anxiolytic compared to the control. While the resulted diving pattern of the MA-treated animals pretreated by RA was novel and different from both the control and MA-treated groups.

Conclusion: The potential of RA combined with a suitable nanoparticle against MA-induced oxidative stress was supported. The high efficiency of ZnO/chitosan in increasing RA penetration to the brain cells was evident. MA at a dose of 25 mg/L is anxiolytic for zebrafish. However, the molecular mechanisms involved in these processes should be studied.

Keywords: Behavioral test; Casp3a; Chitosan nanoparticle; Methamphetamine; Rosmarinic acid; Zebrafish; Zinc oxide.