Therapeutical Potentials of Pomegranate Peel Extract (PPE) in Zebrafish (Danio rerio): Integrated Phytochemical and Neurobehavioral Assessment

ABSTRACT
Objectives: Present study aimed to characterize the phytochemical, anti-inflammatory, antioxidant, antidepressant and
anxiolytic effect of hydroethanolic extract (71.26%) from the peel of Pomegranate (Punica granatum) with In Vivo study
on zebrafish (Danio Rerio).
Methods: A small portion of the dry extract was used for the phytochemical tests for compounds which include tannins,
flavonoids, alkaloids, saponin, and steroids, phenol, Terpenoid. The phosphomolybdenum method was used to assess the
extract's overall antioxidant capability. PPP (Pomegranate Peel Powder) extract was administered orally (n = 10/group) onr
zebrafish (Danio Rerio). The anxiolytic activity was analyzed through the SCOTOTAXIS TEST (light–dark preference
test) test using caffeine as an anxiogenic agent, fluoxetine as a positive control and test sample with 3 doses of PPP extract
(100, 200 & 300 mg/L). The parameters assessed were: Time spent in the white compartment, Latency, Toggle, Erratic
swimming and Number of Freezing. The antidepressant effect was evaluated through the NOVEL TANK test using 1%
ethanol, unpredictable stress, and social isolation as depressors. The parameters assessed were: Time spent at the Top
compartment, Latency, Distance traveled, Number of Freezing and Number of Quadrant crossing. Behavioral analysis was
performed by TOXTRAC software. ELISA is an immunological method which was used to quantify Dopamine, Serotonin
and Cortisol levels in samples of zebrafish.
Results: It showed that the administration of the PPP extract on zebrafish significantly reversed the anxiogenic effect of
caffeine without impairing their locomotion. Additionally, the treatment with PPP extract dosed exerted antidepressant
activity similarly to fluoxetine. The MANOVA test provided definitive statistical evidence of highly significant multivariate
behavioral differences between control and experimental groups (Anxiolytic Behaviour Λ = 0.0209, F = 71.24, p < 0.001
and Depressive Behaviour Λ = 0.0107, F = 114.12, p < 0.001). This represents an exceptionally strong multivariate effect
with 97.91% and 98.93% of variance for Anxiolytic and Depressive Behaviour respectively explained by group
membership.
Conclusion: The animals were active and anxiety / depression-free as a result of the treatment with PPP extract. Also, it
helps to elevate serotonin & dopamine and suppresses cortisol levels in the brains of sample animals. Overall, the results
suggest a significant anxiolytic and antidepressant activity to the PPP extract, which is probably due to the presence of the
major compounds antioxidants and phytochemicals.