Evaluation of Punica granatum fruit juice for anti-anxiety activity

Abstract

Background and Objectives : In traditional system of Indian medicine, Punica granatum L. fruits are widely used for treatment of brain diseases, fever, heart diseases, diarrhea, dysentery, piles, inflammation and bronchitis. However, there is no authentic scientific data reported regarding anxiolytic activity of Punica granatum (Linn) fruit juice. To evaluate the anxiolytic activities of Punica granatum L. fruit juice in various validated animal models of anxiety in mice. Methods: The preliminary phytochemical investigation was carried out with the fruit juice of Punica granatum for qualitative identification of phytoconstituents. Animals were administered with orally 0.1, 0.2, 0.4 and 0.8 ml of Punica granatum fruit juice and observed for its mortality upto 48 hours study period (short term toxicity) to determine LD₅₀. From the LD₅₀ dose, 1/20, 1/10 and 1/5 doses were selected and considered as low, medium and high dose respectively. For assessing the anxiolytic activity, models like Elevated Plus Maze, Hole-Board were used Diazepam was used as a standard reference for anxiolytic. Results: Preliminary phytochemical investigation of the PGFJ ( Punica granatum L. fruit juice) revealed the presence of flavonoids, saponins, tannins, sterols, polyphenols, alkaloids, carbohydrates and proteins. LD₅₀ (acute oral toxicity) of Punica granatum fruit juice was determined in mice with a dose limit of 2000 mg/kg as per OECD guidelines no.425 and even upto 2000 mg/kg dose no mortality was recorded. Hence the experimental doses selected were 1/20, 1/10 and 1/5 of the LD₅₀ value and considered as low, medium and high doses of Punica granatum fruit juice 100, 200 and 400 mg/kg respectively. In Elevated Plus Maze model, low, medium and high doses PGFJ and Diazepam (2 mg/kg) had significantly increased number of entries, time spent in open arms and decreased the number of entries and time spent in closed arms. In Hole- board model, Diazepam (2 mg/kg), medium and high doses (200 and 400mg/kg) but not the low dose (100mg/kg) of PGFJ had significantly increased the number of head dips, latency of first head dip and No.of rearings. Conclusion: The present study ascertains that the plant Punica granatum L. fruit juice possesses significant antianxiety activity in mice and study concludes the beneficial effects of Punica granatum fruit juice in treatment of anxiety signifies the rational basis for its traditional use.