Screening selected medicinal plants for potential anxiolytic activity using an in vivo zebrafish model

Rationale Medicinal plants are used extensively in many countries to treat conditions related to the central nervous system (CNS), and there is renewed interest to explore natural products, which may exhibit CNS activity. Objective In this study, seven plants were selected based on literature reports of their ethnopharmacological use in treating anxiety-related conditions and assayed in a zebrafish model. Methods Crude extracts were prepared with solvents of different polarities, and the maximum tolerated concentration (MTC) of these crude extracts was established. The anxiolytic activity of the crude extracts was determined using 5-day post-fertilization (dpf) zebrafish larvae. General locomotor activity and reverse-thigmotaxis behavior (indicative of anxiolytic activity) were analyzed under continuous illumination and alternating light-dark challenges, which induced anxiety in the zebrafish larvae. Results Of the 28 extracts tested, 13 were toxic according to the MTC values obtained. Larvae were subsequently treated with the 15 non-toxic extracts, at a dose determined in the MTC assay or with 1% DMSO as control. The anxiolytic activity (reverse-thigmotaxis) was demonstrated by an increase in the percentage time spent by the larvae in the central arena of the well. Of the 15 non-toxic extracts tested, the Sceletium tortuosum water extract exhibited the most promising anxiolytic activity. Conclusions The zebrafish model was effective in studying anxiety-related behavior. Thus, the study confirmed that S. tortuosum mitigates anxiety in zebrafish larvae, a step towards the full in vivo validation of the traditional use of the plant.