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

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.