Shivaraj Kulkarni et al / International Journal of Biomedical and Advance Research 2016; 7(9): 452-455. 452 IJBAR (2016) 07 (09) www.ssjournals.com International Journal of Biomedical and Advance Research ISSN: 2229-3809 (Online); 2455-0558 (Print) Journal DOI: 10.7439/ijbar CODEN: IJBABN Original Research Article Evaluation of Punica granatum fruit juice for anti-anxiety activity Shivaraj Kulkarni *1 , Ritesh Bathe 2 and Akshay Javalgikar 2 1 Department of Pharmacology, N.E.T. Pharmacy College, Raichur, India 2 Department of Pharmacology, Sahydri College of Pharmacy, Methwade, India *Correspondence Info: Mr. Shivaraj Kulkarni Department of Pharmacology, Sahyadri College of Pharmacy, Methwade, India E-mail: shivaraj3286@gmail.com 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 LD 50 . From the LD 50 dose, 1/20, 1/10 & 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. LD 50 (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 LD 50 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 number 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. Keywords: Punica granatum, anxiety, Diazepam, Elevated Plus Maze, Hole- board model. 1. Introduction In recent years, anxiolytic drugs have been among the front-runners in terms of the number of prescription written in medical practice. This may be due to the tense life style imposed on man by the competitive atmosphere. Some degree of anxiety is a part of normal life, but treatment needed when it is disproportionate to the situation and excessive.[1] Anxiety disorders may be due to life experiences, heredity and brain chemistry. Chemical messenger’s (neurotransmitters) in the brain regulate thought and feeling. Sometimes has a problem with brain messages sent out because of a chemical imbalance. Approximately 4- 6% of the population suffers from anxiety so severely, that it disrupts routine life functions.[2] In today’s world of stressful life style with the individuals, anxiety is one of the common symptoms associated, affecting almost 1/8 th of the total population worldwide and has became a very important area of research interest in psychopharmacology during this decade and is also obvious component of many psychiatric and medical conditions.[3] Currently, the most widely prescribed medications for anxiety disorders are the Benzodiazepines. However, the clinical uses of Benzodiazepines are limited by their side effects such as psychomotor impairment, potentiation of other central depressant drugs and dependence liability. Therefore, the development of new herbal medications possessing anxiolytic effect without the complication of BZD would be of great importance in the treatment of anxiety-related disorders. In recent years, interest is growing in alternative medicines that include plant derived medications, polyherbal formulations that affect the mind. During last few years there has been increase in usage of alternative medicines by the patients for