Protective effect of pomegranate seed oil against H2O2 -induced oxidative stress in cardiomyocytes

Document Type : Original Research Article


1 Department of Pharmacology, Faculty of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran

2 Pharmacological Research Center of Medicinal Plants, Faculty of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran


Objective: It has been well documented that oxidative stress is involved in the pathogenesis of cardiac diseases. Previous studies have shown that pomegranate seed oil (PSO) has antioxidant properties. This study was designed to investigate probable protective effects of PSO against hydrogen peroxide (H2O2)-induced damage in H9c2 cardiomyocytes.

Materials and Methods: The cells were pretreated 24 hr with PSO 1 hr before exposure to 200 µM H2O2. Cell viability was evaluated using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium (MTT) assay. The level of reactive oxygen species (ROS) and lipid peroxidation were measured by fluorimetric methods.

Results: H2O2 significantly decreased cell viability which was accompanied by an increase in ROS production and lipid peroxidation and a decline in superoxide dismutase activity. Pretreatment with PSO increased viability of cardiomyocytes and decrease the elevated ROS production and lipid peroxidation. Also, PSO was able to restore superoxide dismutase activity.

Conclusion: PSO has protective effect against oxidative stress-induced damage in cardiomyocytes and can be considered as a natural cardioprotective agent to prevent cardiovascular diseases.



Main Subjects

Balasundram N, Sundram K, Samman S. 2006. Phenolic compounds in plants and agri-industrialby-roducts: antioxidant  activity,  occurrence  and potential uses. Food Chem, 99: 191-203.
Boroushaki MT, Arshadi D, Jalili-Rasti H, Asadpour E, Hosseini A. 2013. Protective Effect of Pomegranate Seed Oil Against Acute Toxicity of Diazinon in Rat Kidney. Iran J Pharmaceutic Res, 12: 821-827.
Boroushaki MT, Mollazadeh H, Rajabian A, Dolati K, Hoseini A, Paseban M and Farzadnia M. 2014. Protective effect of pomegranate seed oil against mercuric chloride-induced nephrotoxicity in rat. Ren Fail, 36: 1581-6.
Bouroshaki MT, Sadeghnia HR, Banihasan M, Yavari S. 2010. Protective effect of pomegranate seed oil on hexachlorobutadiene-induced nephrotoxicity in rat kidneys. Ren Fail, 32: 612-617.
Boussetta T, Raad H, Lettéron P, Gougerot-Pocidalo MA, Marie JC, Driss F. 2009. Punicic acid a conjugated linolenic acid inhibits TNFalpha - induced neutrophil hyperactivation and protects from experimental colon inflammation in rats. PLoS One, 4: e6458.
Braga LC, Shupp JW, Cummings C. 2005. Pomegranate extract inhibits Staphylococcus aureus growth and subsequent enterotoxin production. J Ethnopharmacol, 96:335–339.
Brian KM, Strohacker KA, Kueht ML. 2009. Pomegranate seed oil consumption during a period of high-fat feeding reduces weight gain and reduces type 2 diabetes risks in CD-1 mice. Br J Nutr, 102: 54-59.
Caligiani A, Bonzanini F, Palla G, Cirlini M, Bruni R. 2010. Characterization of a potential nutraceutical ingredient: Pomegranate (Punicagranatum L.) seed oil unsaponifiable fraction. Plant Food Hum Nutr, 65: 277-283.
Dhalla NS, Temsah RM, Netticadan T. 2000.Role of oxidative stress in cardiovascular diseases. J. Hypertension, 18: 655–73.
El-Sherbini GM, Ibrahim KM, El Sherbiny ET, Abdel-Had NM, Morsy TA. 2010. Efficacy of Punicagranatumextract on in-vitro andin-vivo control of Trichomonas vaginalis. J Egypt Soc Parasitol, 40:229–244.
Fadavi A, Barzegar M, Azizi MH. 2006. Determination of fatty acids and total lipid content in Oil seed of 25 pomegranates varieties grown in Iran. J Food Comp Anal, 19: 676-680.
Ghorbani A,Hadjzadeh MR, Rajaei Z, Zendehbad SB. 2014. Effects of fenugreek seeds on adipogenesis and lipolysis in normal and diabetic rat. Pak J Biol Sci, 17: 523-528.
Forouzanfar F, Goli AA, Assadpour E, Ghorbani A, Sadeghnia HR. 2013. Protective effect of Punicagranatum L. against serum/glucosedeprivation-induced PC12 cells injury. Evid based Complement Altern Med, 2013: 7167730.
Hartman RE, Shah A, Fagan AM. 2006.  Pomegranate juice decreases amyloid load and improves behavior in a mouse model of Alzheimer’s disease. Neurobiol Dis, 24:506–515.
Higashi Y, Noma K, Yoshizumi M, Kihara Y. 2009.Endothelial function and oxidative stress in cardiovascular diseases. Circ J, 73: 411–418.
Hwang DM, Kundu JK, Shin JW, Lee JC, Lee HJ and Surh YJ. 2007. cis-9, trans-11-Conjugated linoleic acid down-regulates phorbol ester-induced NF-𝜅B activation and subsequent COX-2 expression in hairless mouse skin by targeting I𝜅B kinase and PI3K-Akt. Carcinogenesis,  28: 363–371.
Kullkarni AP, Aradhya SM. 2005.Chemical Changes and Antioxidant Activity in Pomegranate Arils during Fruit Development. Food Chem, 93: 319-324.
Lansky EP, Newmana RA. 2007. Punicagranatum (pomegranate) and its potential for prevention   and   treatment    of    inflammation   and cancer:  Review.  J  Ethnopharmacol, 109: 177-206.
L'Ecuyer T, Allebban Z, Thomas R, Vander Heide R.  2004. Glutathione S-transferase overexpression protects against anthracycline-induced H9c2 cell death. Am. J Physiol Heart Circ Physiol, 286: H2057–H2064.
Miguel G, Dandlen S, Antunes D, Neves A, Martins D. 2004. The Effect of Two Methods of Pomegranate (Punicagranatum L) Juice Extraction on Quality during Storage at 4°C. J Biomed Biotech, 5: 332-337.
Miguel G, Fontes C, Antunes D, Neves A, Marthins D. 2004. Anthocyanin Concertration of Assaria Pomegranate Fruits during Different Cold Storage Conditions. J Biomed Biotech, 5: 338-342.
Mortazavian SM, Ghorbani A, Hesari TG. 2012. Effect of hydro-alcoholic extracts of viola tricolor and its fractions on proliferation of cervix carcinoma cells. Iran J Obstet Gynecol Infertility, 15: 9-16.
Nagao K, Yanagita T. 2005.Conjugated fatty acids in food and their health benefits. J Biosci Bioengineering, 100: 152–157.
Noda Y, Kaneyuki T, Mori A, Packer L. 2002. Anitioxdant Activities of Pomegranate Fruit Extract and ItsAnthocyanidins:  Delphinidin, Cyanidin, and Pelargonidin. J Agric Food Chem, 50: 166-171.
Okamoto JM, Hamamoto YO, Yamato H, Yoshimura H. 2004. Pomegranate Extract  Improves  a  Depressive  State  and Bone  Properties  in  Menopausal  Syndrome Model  Ovariectomized  Mice.  J Ethnopharmacol, 92: 93-101.
Ramanathan L, Das NP. 1992. Inhibitory effects of some natural products on metal-induced lipid oxidation in cooked fish. Biol Trace Elem Res, 34:35–44.
Sadeghian A, Ghorbani A, Mohamadi-Nejad A, Rakhshandeh H. 2011. Antimicrobial activity of aqueous and methanolic extracts of pomegranate fruit skin. Avicenna J Phytomed, 1: 67-73.
Sadeghnia HR, Yousefsani BS, Rashidfar M, Boroushaki MT, Assadpour E, Ghorbani A. 2013.Protective effect of rutin on hexachlorobutadiene-induced nephrotoxicity. Ren Fail, 35: 1151-1155.
Saha SS, Ghosh M. 2009. Comparative study of antioxidant activity of alpha-eleostearic acid and punicic acid against oxidative stress generated by sodium arsenite. Food Chem Toxicol, 47: 2551-2556. 
Saha SS, Dasgupata  P, Sengupata S, Ghosh M. 2012. Synergistic effect of conjugated linolenic acid isomers against induced oxidative stress, inflammation and erythrocyte membrane disintegrity in rat model. Biochim Biophys Acta, 1820:1951–1970.
Sassano G, Sanderson P, Franx J, Groot P, Straalen JV, Bassaganya-Riera J. 2009. Analysis of pomegranate seed oil for the presence of jacaric acid. J Sci Food Agr, 89: 1046-1052.
Singh RP, Murthy KNC, Jayaprakasha GK. 2002. Studies on the Antioxidant Activity of Pomegarante (Punicagranatum) Peel and Seed Extracts Usning in Vitro Models. J Agric Food Chem, 50: 81-86.
Sun B, Sun GB, Xiao J, Chen RC, Wang X, Wu Y, Cao L, Yang ZH, Sun XB. 2012. Isorhamnetin inhibits H₂O₂-induced activation of the intrinsic apoptotic pathway in H9c2 cardiomyocytes through scavenging reactive oxygen species and ERK inactivation. J Cell Biochem, 113: 473-485.
Tsutsui H, Kinugawa S, Matsushima S. 2011.Oxidative stress and heart failure. Am J Physiol, 301: H2181-H90.
Turakhia S, Venkatakrishnan CD, Dunsmore K, Wong H, Kuppusamy P, Zweier JL, Ilangovan G. 2007. Doxorubicin-induced cardiotoxicity: direct correlation of cardiac fibroblast and H9c2 cell survival and aconitase activity with heat shock protein 27. Am J Physiol Heart Circ Physiol, 293: H3111.
Viuda-Martos M, Fernandez-L O, Pereza Alvarez JA. 2010. Pomegranate and its many functional components as related to human health: a review. Comperhensive Reviews in Food Science and Food Safety, 9: 635-654.
Winstead M, Lucas K, Dennis E. 2005. Group IV cytosolic phospholipase A2 mediates arachidonic acid release in H9c2 rat cardiomyocyte cells in response to hydrogen peroxide. Prostagland Other Lipid Med, 78: 55-66.
Xin H, Liu XH, Zhun Zhu Y. 2009. Herbal eonurine attenuates doxorubicin-induced apoptosis in H9c2 cardiac muscle cells. Eur J Pharmacol, 612: 75–79.