Pomegranate peel extract inhibits internalization and replication of the influenza virus: An in vitro study

Document Type : Original Research Article


1 Medical Plants Research Center, Basic Health Sciences Institute, Shahrekord University of Medical Sciences, Shahrekord, Iran.

2 Cellular and Molecular Research Center, Basic Health Sciences Institute, Shahrekord University of Medical Sciences, Shahrekord, Iran.

3 Department of Pathobiology, Infectious Disease and Public Health, School of Animal and Veterinary Sciences, University of Adelaide, Adelaide, Australia.

4 Department of Microbiology, College of Veterinary Medicine, Shahrekord Branch, Islamic Azad University.


Objective: Influenza virus, which is associated with high level of morbidity and mortality, has been recently considered a public health concern; however, the methods of choice to control and treat it are limited. Our previous study showed anti-influenza virus activity of pomegranate peel extract (PPE). In this study, the mechanism through which PPE acts against influenza virus A/Puerto Rico/8/34 (H1N1; PR8) was investigated.
Materials and Methods:Ethyl alcohol extract of pomegranate (Punica granatum L.) peel was prepared, and the action mechanism of PPE in inhibiting influenza replication was studied by time-of-drug-addition assay, virucidal activity, RNA replication, hemagglutination inhibition assay, viral mRNA expression, and western blot analysis.
Results: PPE inhibited viral polymerase activity, viral RNA replication, and viral protein expression but could not affect hemagglutination inhibition and virucidal activity. According to time-of-drug-addition assay results, PPE inhibited the virus adsorption and early steps of influenza replication.
Conclusion: This study demonstrated that the antiviral effect of PPE on influenza virus is most probably associated with inhibition of viral adsorption and viral RNA transcription.


Main Subjects

Abdal Dayem A, Choi HY, Kim YB, Cho SG. 2015. Antiviral effect of methylated flavonol isorhamnetin against influenza. PLoS One, 10: e0121610.
Asmaa MJ, Ali AJ, Farid JM, Azman S. 2015. Growth inhibitory effects of crude pomegranate peel extract on chronic myeloid leukemia, K562 cells. J Appl Basic Med Res, 5: 100-105.
Centers for Disease Control and Prevention. 2018. Influenza antiviral medications: summary for clinicians. Available on: https://www.cdc.gov/flu/professionals/antivirals/summary-clinicians.htm (Accessed on December 10, 2018).
Colombo E, Sangiovanni E, Dell'agli M. 2013. A review on the anti-inflammatory activity of pomegranate in the gastrointestinal tract. Evid Based Complement Alternat Med, 2013: 247145.
Dapat C, Kondo H, Dapat IC, Baranovich T, Suzuki Y, Shobugawa Y, Saito K, Saito R, Suzuki H. 2013. Neuraminidase inhibitor susceptibility profile of pandemic and seasonal influenza viruses during the 2009-2010 and 2010-2011 influenza seasons in Japan. Antiviral Res, 99: 261-269.
Elyatem SM, Kader AA. 1984. Post-harvest physiology and storage behaviour of pomegranate fruits. Sci Hortic, 24: 287-298.
Godornes C, Leader BT, Molini BJ, Centurion-Lara A, Lukehart SA. 2007. Quantitation of Rabbit Cytokine mRNA by Real-Time RT-PCR. Cytokine, 38: 1-7.
Haber SL, Joy JK, Largent R. 2011. Antioxidant and antiatherogenic effects of pomegranate. Am J Health Syst Pharm, 68: 1302-1305.
Haidari M, Ali M, Ward Casscells S, Madjid M. 2009. Pomegranate (Punica granatum) purified polyphenol extract inhibits influenza virus and has a synergistic effect with oseltamivir. Phytomedicine, 16: 1127-1136.
Hayden FG. 2006. Respiratory viral threats. Curr Opin Infect Dis, 19: 169-178.
He W, Han H, Wang W, Gao B. 2011. Anti-influenza virus effect of aqueous extracts from dandelion. Virology J, 8: 538-538.
Howell AB, D'Souza DH. 2013. The Pomegranate: effects on bacteria and viruses that influence human health. Evid Based Complement Alternat Med, 2013: 606212.
Ibrahium M. 2010. Efficiency of pomegranate peel extract as antimicrobial, antioxidant and protective agents. World J Agric Sci, 6: 338-344.
Ismail T, Sestili P, Akhtar S. 2012. Pomegranate peel and fruit extracts: a review of potential anti-inflammatory and anti-infective effects. J Ethnopharmacol, 143: 397-405.
Jackson RJ, Cooper KL, Tappenden P, Rees A, Simpson EL, Read RC, Nicholson KG. 2011. Oseltamivir, zanamivir and amantadine in the prevention of influenza: a systematic review. J Infect, 62: 14-25.
Kim ND, Mehta R, Yu W, Neeman I, Livney T, Amichay A, Poirier D, Nicholls P, Kirby A, Jiang W, Mansel R, Ramachandran C, Rabi T, Kaplan B, Lansky E. 2002. Chemopreventive and adjuvant therapeutic potential of pomegranate (Punica granatum) for human breast cancer. Breast Cancer Res Treat, 71: 203-217.
Kim Y, Narayanan S, Chang KO. 2010. Inhibition of influenza virus replication by plant-derived isoquercetin. Antiviral Res, 88: 227-235.
Kummer S, Flöttmann M, Schwanhäusser B, Sieben C, Veit M, Selbach M, Klipp E, Herrmann A. 2014. Alteration of protein levels during influenza virus H1N1 infection in host cells: a proteomic survey of host and virus reveals differential dynamics. PLoS One, 9: e94257.
Lansky EP, Newman RA. 2007. Punica granatum (pomegranate) and its potential for prevention and treatment of inflammation and cancer. J Ethnopharmacol, 109: 177-206.
Matusevich OV, Egorov VV, Gluzdikov IA, Titov MI, Zarubaev VV, Shtro AA, Slita AV, Dukov MI, Shurygina AP, Smirnova TD, Kudryavtsev IV, Vasin AV, Kiselev OI. 2015. Synthesis and antiviral activity of PB1 component of the influenza A RNA polymerase peptide fragments. Antiviral Res, 113: 4-10.
Moradi MT, Karimi A, Lorigooini Z, Pourgheysar B, Alidadi S, Hashemi L. 2017. In vitro anti influenza virus activity, antioxidant potential and total phenolic content of twelve Iranian medicinal plants. Marmara Pharm J, 21: 843-851.
Mosmann T. 1983. Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods, 65: 55-63.
Ohuchi M, Asaoka N, Sakai T, Ohuchi R. 2006. Roles of neuraminidase in the initial stage of influenza virus infection. Microbes Infec, 8: 1287-1293.
Plumb GW, de Pascual-Teresa S, Santos-Buelga C, Rivas-Gonzalo JC, Williamson G. 2002. Antioxidant properties of gallocatechin and prodelphinidins from pomegranate peel. Redox Report, 7: 41-46.
Reddy MK, Gupta SK, Jacob MR, Khan SI, Ferreira D. 2007. Antioxidant, antimalarial and antimicrobial activities of tannin-rich fractions, ellagitannins and phenolic acids from Punica granatum L. Planta Med, 73: 461-467.
Reed LJ, Muench H. 1938. A simple method of estimating fifty per cent endpoints. Am J Epidemiol, 27: 493-497.
Shapiro GI, Gurney T, Jr Krug RM. 1987. Influenza virus gene expression: control mechanisms at early and late times of infection and nuclear-cytoplasmic transport of virus-specific RNAs. J Virol, 61: 764-773.
Sugita Y, Sagara H, Noda T, Kawaoka Y. 2013. Configuration of Viral Ribonucleoprotein complexes within the influenza A virion. J Virol, 87: 12879-12884.
van der Vries E, Schutten M, Fraaij P, Boucher C, Osterhaus A. 2013. Influenza virus resistance to antiviral therapy. Adv Pharmacol, 67: 217-246.
World Health Organization (WHO). 2011. Global Influenza Surveillance Network. Manual for the laboratory diagnosis and virological surveillance of influenza. Available from: http://whqlibdoc. who.int/publications/2011/9789241548090_eng.pdf (Last access: 06/25/2014).
Zarubaev VV, Garshinina AV, Tretiak TS, Fedorova VA, Shtro AA, Sokolova AS, Yarovaya OI, Salakhutdinov NF. 2015. Broad range of inhibiting action of novel camphor-based compound with anti-hemagglutinin activity against influenza viruses in vitro and in vivo. Antiviral Res, 120: 126-133.