Author(s): Anil Savali, Mulchand Shende, S. B. Chandrasekhar


DOI: 10.52711/0974-360X.2023.00771   

Address: Anil Savali1, Mulchand Shende2*, S. B. Chandrasekhar3
1Pharmacy Department, Government College of Pharmacy, Vidyanagar, Karad, Maharashtra, India, 415124.
2Department of Pharmaceutics, Government College of Pharmacy, Vidyanagar, Karad, Maharashtra, India, 415124.
3Drug Testing Laboratory, Drugs Control Department, Palace Road, Bangalore, 560001.
*Corresponding Author

Published In:   Volume - 16,      Issue - 10,     Year - 2023

The memory loss associated with increased oxidative stress due to increased level of free radicals. Oxidative stress has been involved in several diseases includes cancer, atherosclerosis and neurodegenerative diseases.This study aimed to estimate the correlation of antioxidant and antistress activity with nootropic activity of aqueous extract of pomegranate (Punica granatum) by in-vivo noninvasive studies using rats. The in-vitro antioxidant activity was determined based on the ability of the Punica granatum to scavenge free radicals and lipid peroxidation inhibiting activity. The antistress effect of the aqueous extract of Punica granatum for 24h treatment (100 and 300mg/kg, p.o.) was evaluated by using the forced swim stress test in rats. The 24h urinary excretion of vanillylmandellic acid (VMA), 5-hydroxyindoleacetic acid (5-HIAA), homovanillic acid (HVA) and ascorbic acid (AA) was determined in all groups under normal and stressed conditions by HPLC and spectrophotometric methods in all groups, were selected as noninvasive biomarkers. Nootropic activity activities of Punica granatum fruit extract were estimated as locomotor and working memory in rats in a Y-maze apparatus. Administration of aqueous extract of Punica granatum at a dose of 100 and 300mg/kg reduced the urinary metabolite levels. Punica granatum treatment showed significant dose dependent variation in non-invasive biomarker levels in urine samples of rats taken after 24h. The treatment with Punica granatum extracts improved the percentage entry of rats into safer zone, it means acquisition (learning) retention and retrieval (memory) were improved in rats compared to stress controls. Cognition determined by working memory and locomotor activity results were shown to be dose-dependent. The results of this study strongly suggestedantioxidant, antistress and nootropic activity effect of Punica granatumin rodents were correlated. The data obtained were analyzed by one-way ANOVA followed bytukey test. p<0.05 was considered to be significant. There is substantial evidence that flavonoids play an active role in providing antioxidant, antistress and nootropic activities of Punica granatum extracts. The findings of the present investigations indicate that the Punica granatum has significant antistress activity, which may be due to the immunostimulating property and increased resistance, nonspecifically, against all experimental stress conditions. The study provided scientific evidence for their utility as nootropic agents and to advocate their use in foods.

Cite this article:
Anil Savali, Mulchand Shende, S. B. Chandrasekhar. Antioxidant, Antistress, Nootropic activity and its Correlation studies of aqueous extract of Punica granatum fruit estimated by Noninvasive biomarkers and Y-maze test in rodents. Research Journal of Pharmacy and Technology 2023; 16(10):4751-8. doi: 10.52711/0974-360X.2023.00771

Anil Savali, Mulchand Shende, S. B. Chandrasekhar. Antioxidant, Antistress, Nootropic activity and its Correlation studies of aqueous extract of Punica granatum fruit estimated by Noninvasive biomarkers and Y-maze test in rodents. Research Journal of Pharmacy and Technology 2023; 16(10):4751-8. doi: 10.52711/0974-360X.2023.00771   Available on:

1.    Seyle H. The evolution of stress concept. Am Sci. 1973; 61:692-9.
2.    Gerard E, Konrad B, Colin L, Jean-Marie M. Prospects for pharmacological intervention in Alzheimer disease. Neurological Review. 2000; 57(4):454-459. doi:10.1001/archneur.57.4.454
3.    Perrig WJ, Perrig P, Stahelin HB. The relation between antioxidants and memory performance in the old and very old. Journal of the American Geriatrics Society. 1997; 45(6):718-24. doi: 10.1111/j.1532-5415.1997.tb01476.x
4.    Rahman T, Hosen I, Islam T, Shekhar HU. Oxidative stress and human health. Advances in Biosciences and Biotechnology. 2012; 3(7A):997-19.
5.    Kumar A, Rinwa P, Kaur G, Machawal L. Stress: Neurobiology, consequences and management. Journal of Pharmacy and Bioallied Sciences. 2013; 5(2):91-7. doi: 10.4103/0975-7406.111818
6.    Rogers SH, Farlow MR, Doody RS, Mohs R, Friedhoff, LI. A 24-week, double-blind, placebo-controlled trial of donepezil in patients with Alzheimer's disease. Donepezil Study Group. Neurology. 1998 ; 50(1): 136-45. doi: 10.1212/wnl.50.1.136
7.    McCarty R. Stress-Neurochemical and humoral mechanism. In: Van Loon GR, McCarty RG, editors. Stress research: Principles, problems, and prospects. New York: Breach Science Publishers; 1987; pp. 3–13.
8.    Piato AL, Detanico BC, Jesus JF, Lhullier FL, Nunes DS, Elisabetsky E. Effects of Marapuama in the chronic mild stress model: Further indication of antidepressant properties. Journal ofEthnopharmacology. 2008; 118(2):300-4. doi: 10.1016/j.jep.2008.04.018. Epub 2008 Apr 24
9.    Kirtikar KR, Basu BD, ICS. In: Indian medicinal plants.  Blatter E, Caius JF, Mhaskar KS, editors. Vol. 3. Allahabad, India: Published by Lalit Mohan Basu. 1981; 2nd ed: pp. 1707–8.
10.    Galani VJ, Patel BG, Patel NB. Argyreia speciosa (Linn.f.) sweet: A comprehensive review. Pharmacognosy Reviews. 2010; 4(8):172–8. doi: 10.4103/0973-7847.70913
11.    Camposa MM, Fernandesa ES, Ferreiraa J, Bortolanzaa LB, Adair ARS, Calixto JB. Pharmacological and neurochemical evidence for antidepressant-like effects of the herbal product Catuama. Pharmacology, Biochemistry and Behavior. 2004; 78(4):757-64. doi: 10.1016/j.pbb.2004.05.024
12.    Middha SK, Usha T, Pande V. HPLC Evaluation of Phenolic Profile, Nutritive Content, and Antioxidant Capacity of Extracts Obtained from Punica granatum Fruit Peel. Advances in Pharmacological Sciences. 2013: 1-6. doi:
13.    Vignesh Balaji. E, Tamil Selvan. A, Srinivasan. A, Nandhini. S. Punica granatum root (s): Phytocompounds analysis, Anti-oxidant and Anti-microbial activity. Asian Journal of Pharmaceutical Analysis. 2019; 9(3):123-127. doi: 10.5958/2231-5675.2019.00023.1
14.    Ranjana Himalian, Mahendra P. Singh. A Comparative account on Antioxidant activities, Total phenolic and Flavonoid contents of Punica granatum, Carica papaya, Foeniculumvulgare, Trigonellafoenum-graecum, and Urticadioica: An in-vitro Evaluation. Research Journal of Pharmacy and Technology. 2022; 15(3):1175-3. doi: 10.52711/0974-360X.2022.00197
15.    Punasiya R, Joshi A, Yadav S, Patidar K, Kapse K. Antimicrobial Activity of the Leaves Extract of Punica granatum Linn. Research Journal of Pharmacognosy and Phytochemistry. 2010; 2(1): 21-24.
16.    Punasiya R, Joshi A, Patidar K. Antidiabetic Effect of an Aqueous Extract of Pomegranate (Punica granatum L.) Peels in Normal and Alloxan Diabetic Rats. Research Journal of Pharmacy and Technology. 2010; 3(1): 272-274.
17.    Rajasekhar Pinnamaneni. Preliminary Phytochemical Investigation of peel of Pomegranate Punica granatum L. Research Journal of Pharmacy and Technology. 2018; 11(8): 3609-3613. doi: 10.5958/0974-360X.2018.00664.9
18.    Hartman RE, Shah A, Fagan AM, Schwetye KE, Parsadanian M, Schulman RN, Finn MB, Holtzman DM. Pomegranate juice decreases amyloid load and improves behavior in a mouse model of Alzheimer’s disease. Neurobiology of Disease. 2006; 24(3): 506-15. doi: 10.1016/j.nbd.2006.08.006
19.    Aviram M, Volkova N, Coleman R, Dreher M, Reddy MK, Ferreira D, Rosenblat M. Pomegranate phenolics from the peels, arils, and flowers are antiatherogenic: studies in-vivo in atherosclerotic apolipoprotein E-deficient (E°) mice and in-vitro cultured macrophages and lipoproteins. Journal of Agriculture and Food Chemistry. 2008; 56(3):1148-157. doi:10.1021/jf071811q
20.    Hartlapp I, Abe R, Saeed RW, Peng T, Voelter W, Bucala R, Metz CN. Fibrocytes induce an angiogenic phenotype in cultured endothelial cells and promote angiogenesis in-vivo. Federation of American Societies for Experimental Biology Journal. 2001; 15(12): 2215-24. doi: 10.1096/fj.01-0049com
21.    Valadares MC, Perira ERT, Benfica PL, Paula JR. Assessment of mutagenic and antimutagenic effects of Punica granatum in mice. Brazilian Journal of Pharmaceutical Sciences. 2010:46(1):121-7.
22.    Kokate CK. Practical Pharmacognosy. New Delhi, India: Vallabh Prakashan. 1994; 4thed: pp. 107-109.
23.    McCord JM, Fridovich I. The Utility of Superoxide Dismutase in Studying Free Radical Reactions. The Journal of Biological Chemistry. 1969; 244(22): 6056-63.
24.    Elizabeth ZI, Rao MNA. Oxygen radical scavenging activity of curcumin. International Journal of Pharmaceutics. 1990; 58(3): 237–40. 10.1016/0378-5173(90)90201-E
25.    Ohkawa H, Ohishi N, Yagi K. Assay for lipid peroxides in animal tissues by thiobarbituric acid reaction. Analytical Biochemistry. 1979; 95(2):351-8. doi: 10.1016/0003-2697(79)90738-3.
26.    Roe JH, Kuether CA. The determination of ascorbic acid in whole blood and urine through the 2, 4-dinitrophenylhydrazine derivative of dehydroascorbic acid. Journal of Biological Chemistry. 1943; 147:399-07.
27.    Onaolapo OJ, Onaolapo AY, Mosaku TJ, Akanji OO, Abiodun OR. Elevated Plus maze and Y-maze Behavioural effects of Subchronic, Oral Low Dose Monosodium Glutamate in Swiss Albino Mice. Journal of Pharmacy and Biological Sciences. 2012; 3(4): 21-7.
28.    Pisano JJ, Crout JR, Abraham. Determination of 3-Methoxy, 4-hydroxymandellic acids in urine. Clinica Chimica Acta.  1962; 7:285-91. doi: 10.1016/0009-8981(62)90022-0
29.    Kagedal B, Goldstein DS. Catecholamines and their metabolites. Journal of Chromatography. 1988; 429:177-223. doi: 10.1016/s0378-4347(00)83871-2
30.    Rajasekaran A, Sivagnanam G, Xavier R. Nutraceuticals as therapeutic agents: A Review. Research Journal of Pharmacy and Technology. 2008; 1(4): 171-174.
31.    Yoshikawa T, Naito Y, What is Oxidative Stress? The Japanese Association of Medical Sciences. 2002; 45(7):271–6.
32.    Ghanshyam B. Jadhav, Ravindranath B. Saudagar. Free radical Scavenging and Antioxidant Activity of Punica granatum Linn. Asian Journal of Research in Pharmaceutical Sciences. 2014; 4(2): 51-4.
33.    Andersen JK. Oxidative stress in neurodegeneration: Cause or consequence? Nature Medicine. 2004; 10: S18-25. doi: 10.1038/nrn1434
34.    Madhu C, Swapna J, Neelima K, Shah MV. A comparative evaluation of the antioxidant activity of some medicinal plants popularly used in India. Asian Journal of Research in Pharmaceutical Science. 2012; 2(3):98-100.
35.    Gregory S, Kelly ND. Nutritional and botanical interventions to assist with adaptation to stress. Alternative Medicine Review. 1999; 4(4): 249-65.
36.    Kvetnansky R, Sun CL, Lake CR, Thoa N, Torda T,Kopin IJ. Effect of handling and forced immobilization on rat plasma levels of epinephrine, norepinephrine and dopamine-beta-hydroxylase. Endocrinology.1978; 103(5):1868-74. doi: 10.1210/endo-103-5-1868
37.    Desole MS, Esposito G, Fresu L, Migheli R, Sircana S, Delogu R, Miele M, Miele E. Allopurinol protects against manganese-induced oxidative stress in the striatum and in the brainstem of the rat. Neuroscience Letters. 1995; 192(2):73-6. doi: 10.1016/0304-3940(95)11613-2
38.    Platzer C, Docke W, Volk H, Prosch S. Catecholamine trigger IL-10 release in acute systemic stress reaction by direct stimulation of its promoter enhancer activity in monocytic cells. Journal of Neuroimmunology. 2000; 105(1):31-8. doi: 10.1016/s0165-5728(00)00205-8
39.    Axelord J, Reisine TD. Stress hormones: Their interaction and regulation. Science. 1984; 224(4648): 452-9. doi: 10.1126/science.6143403.
40.    Sreemantula S, Nammi S, Kolanukonda R, Koppula S, Boini KM. Adaptogenic and nootropic activities of aqueous extract of Vitisvinifera (grape seed): an experimental study in rat model. BMC Complementary and Alernative Medicine. 2005; 5(1):1-8. doi:10.1186/1472-6882-5-1
41.    Lupien SJ, Maheu F, Tu M, Fiocco A and Schramek TE. The effects of stress and stress hormones on human cognition: Implications for the field of brain and cognition. Brain and Cognition. 2007; 65(3):209-37. doi: 10.1016/j.bandc.2007.02.007
42.    Carrasco GA, Van de Kar LD. Neuroendocrine pharmacology of stress. European Journal of Pharmacology. 2003; 463:235-72. doi: 10.1016/s0014-2999(03)01285-8

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