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1- Department of Biology, Faculty of Basic Sciences, University of Zabol, Bonjar Road, Zabol, Iran.
Abstract:  
Background: Methamphetamine (MA) acts as a powerful oxidant and Rosmarinic acid (RA) is a powerful herbal antioxidant. Oxidative stress mediated by MA results in apoptosis and caspase-3 is one of the key enzymes in the apoptosis process. MA is able to 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 has been demonstrated in zebrafish.
Purpose: This study aims to investigate MA ability in imposing the brain cell to increase the synthesis of casp3a mRNA in zebrafish brain tissue. We hypothesized that a powerful antioxidant such as RA may be able to counteract MA-mediated oxidative stress and its consequences. For better penetration of RA across the blood brain barriers (BBBs), RA was combined with ZnO/chitosan nanoparticles. To evaluate whether MA with the concentration of 25 mg/l is anxiogenic or anxiolytic and the effects of RA on it, one-sided trapezoidal tank diving tests were applied for 3 groups of MA-treated, MA & RA-treated and saline as the control group.
Study design: 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 cotreated with RA, MA cotreated 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.
Methods: MA solution was prepared with the concentration of 25 mg/l and RA solution was prepared by DPPH test with the antioxidant power about 97%. Each solution was administered by immersing 20 zebrafish for 20 minutes, once per day for a continuous 7 days. The level of casp3a mRNA was quantified by making use of 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. By the diving test of MA-treated fishes, MA found to be anxiolytic compared to the control. While the resulted diving pattern of the MA-treated animals pre-treated by RA was novel and different than both the control and MA-treated groups.
Conclusion: The potential of RA combined to 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. 25 mg/l of MA for zebrafish is anxiolytic. However, the molecular mechanisms involved in these processes are needed to be studied.
Type of Study: Original | Subject: Behavioral Neuroscience
Received: 2019/04/10 | Accepted: 2019/12/1

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