Antiviral Activity of Polyphenols Extracts From Daucus carota against Herpes Simplex Virus type 1

Abstract

Herpes Simplex Virus type 1 (HSV-1), is a common and recurrent human virus with no medical cure. Polyphenols, on the other hand, are known to exhibit very strong oxygen scavenging properties; therefore they are considered a very rich source of antioxidants. In this research, the effect of extraction method using different solvents on the content of polyphenols extracted from the leaves and roots of D.carota was studied, and their relationship with antioxidant activity was investigated. The examined extracts were tested for cytotoxicity on Vero cell line, with reference to IC50, and other non-toxic concentrations of all the extracts. The antiviral activity against HSV-1 for all non-toxic concentrations of the extracts was determined using plaque reduction assay, which revealed that the inhibitory activity of the HSV-1 virus was dose dependent on the polyphenol content of the examined extracts. The MIC for all the extracts was also determined as well as the EC50 and the SI for all the extracts. Calculated SI values for the examined extracts showed promising SI on the aqueous extracts of the D.carota roots and leaves, and hence can be used as therapeutic medication for HSV-1. Direct contact between the HSV-1 and the examined extracts in cell-free assay system showed different degrees of virucidal activity depending on the polyphenolic content of these extracts. In order to study other possible mode of action, Vero cells were treated with the examined extracts before, during, and after virus infection to give an insight on the interference of the extract in each step in the virus life cycle. Examined extracts exhibited the antiviral activity against HSV-1 via blocking of the virus attachment and penetration and inhibition of the early stage of viral replications

Keywords

• antiviral activity, HSV-1, polyphenols

References

1. Amoros, M., E. Lurton, J. Boustie and L. Girre, (1994). comparison of the anti-herpes simplex virus activities of propolis and 3-methyl-But- 2Enyl cafferate., J. of Natural products., 57: 644-647.
2. Arthanari, S.K., Vanitha, J., Ganesh, M., and Venkateshwaran, K., (2012). Evaluation of antiviral and cytotoxic activities of methanolic extract of S.grandiflora (Fabaceae) flowers. Asian Pacific Journal of Tropical Biomedicine. V2, issue, 2
3. Block, G. (1994). Nutrient sources of provitamin A carotenoids in the American diet. American Journal of Epidemiology 139:290-293.
4. Ciz, M., Pavelkova, M., Gallova, L., Kralova, J., Kubala, L., and Lojek, A., (2008). The influence of wine polyphenols on reactive oxygen and nitrogen species production by murine macrophages RAW 264.7. Physiol Res 57: 393–402.
5. Collier, L., and Oxford, J. (2000). The herpesviruses: general properties. In: Human virology, 2nd ed, Oxford University Press, pp. 131-140.
6. Dalva, A., Rosângela, P., José Ricardo, P., and Jorge, M.,(2009). Inhibition of DNA Virus: Herpes-1 (HSV-1) in cellular culture replication, through an antioxidant treatment extracted from rosemary spice. Brazilian Journal of Pharmaceutical Sciences. vol. 45, n. 1.
7. Danaher, R.J., Wang, C., Daj, J., Mumper, R.J., and Miller, C.S., (2011). Antiviral effects of blackberry extract against herpes simplex virus type 1. Oral Surg ; 112(3).
8. Djeridane, A., Yousfi, M., Nadjemi, B., Boutassouna, D., Stocker, P., Vidal, N.,(2006). Antioxidant activity of some Algerian medicinal plants extracts containing phenolic compounds. Food Chemistry 97:654– 660

9. Donglin, Z., and Yasunori, H., (2004). Phenolic compounds and their antioxidant properties in different tissues of carrots (Daucus carota L.).
10. Gebicka, L., Banasiak, E. (2009). Flavonoids as reductants of ferryl hemoglobin. Acta Biochim Pol 56: 509– 513.
11. Katalinic, V., Milos, M., Kulisic, T., and Jukic, M. (2006). Screening of 70 medicinal plants for antioxidant capacity and total phenols. Food Chem 94: 550–557.
12. Khan, M.T., Ather, A., Thomson, K.D., and Gambari, R. (2005). Extracts and molecules from medicinal plants against herpes simplex viruses. Antiviral. Res. 67: 107-119.
13. Kratchanova, M., Denev, P., Ciz, M., Lojek, A., and Mihailov, A., (2010). Evaluation of antioxidant activity of medicinal plants containging polyphenol compounds. Comparison of two extraction systems. Acta Biochemica Polonica ; 57:229-234.
14. Kratz, J.M., Andrighetti-Fröhner, C.R., Kolling, D.J., Leal1, P.C., Santos, C.C., Yunes1, R.A., Nunes1, R.J.,
15. Trybala, E., Bergström, T., Frugulhetti, I.C., Barardi, C.R., and Simões, C.M.O., (2008). Anti-HSV-1 and anti-HIV-1 activity of gallic acid and pentyl gallate. Mem Inst Oswaldo Cruz, Rio de Janeiro, Vol. 103(5): 437-442
16. Ojo, O.O., Oluyege, J.O., and Famurewa (2009). Antiviral properties of two Nigerian plants. African Journal of Plant Sciences, 3(7):157-159.
17. Pujol, C.A., Estevez, J.M., Carlucci, M.J., Ciancia, M., Cerezo, A.S., and Damonte, E.B., (2002). Novel DLgalactan hybrids from the red seaweed Gymnogongrus torulosus are potent inhibitors of herpes simplex virus and dengue virus. Antimicrob. Agents Chemother., v.13, n.2, p.83-89. 1
18. Raenu, Y., Songyot, A., and Yingmanee, T., (2010). Anti-herpes simplex virus activity of extracts from the culinary herps Ocimum sanctum L., Ocimum bascilicum L. African Jotnal of Biotechnology Vol.10(5), pp. 860-866.
19. Raenu, Y., Sunee, C., and Yingmanee, T.,(2011). Inhibitory effect of aromatic herbs, lavender, sage and chamomile against herpes simplex virus infection. Jotnal of Biotechnology Vol.10(68), pp.15394- 15401.
20. Rice, E. C.A., Miller, N.J., Bolwell, P.G., Bramley, P.M. and Pridham, J.B. (1995). The relative antioxidant activities of plant-de-rived polyphenolic flavonoids. Free Radical Research 22:375-383
21. Sayed, H.M., Mahmoud, S.S., Hossein, K., and Farah, B.S., (2012). Evaluation of in vitro antiviral activity of Chelidonium majus L. against herpes simplex virus type-1. African Journal of Microbiology Research Vol. 6(20), 4360-4364.
22. Schuhmacher, A., J. Reichling, and P. Schnitzler. (2003). Virucidal effect of peppermint oil on the enveloped viruses herpes simplex virus type 1 and type 2 in vitro. Phytomedicine. 6: 504-510.
23. Simões, C.M., Amoros, M., and Girre, L., (1999). Mechanism of antiviral activity of triterpenoid saponins. Phytother Res 13: 323-328.
24. Singleton, V.L. and Rossi, J.A. (1965). Colorimetry of total phenolics with phosphomolybdicphosphotungstic aci+d reagents. Am. J. Enol. Vitic. 16, 144-158.
25. Slinkard, K. and Singleton, V.L. (1977). Total phenol analysis: automation and comparison with manual Methods, Am. J. Enol. Vitic. 28, 49-55.
26. Taga, M.S., Miller, E.E., and Pratt, D.E., (1984). Chia seed as a source of natural lipid antioxidants. JAOCS 61:928-931.
27. Trouong, V.D., Mcfeeters, R.F., Thompson, R.T, Dean, L.L., and Shofran, B., (2011). Phenolic acid content and composition in leaves and roots of common commercial sweetpotato (Ipomea batatas L.) cultivars in the United States. Food Chemistry and Toxicology.
28. Urrea, D., Eim, V.S., Gonzalez-Centeno, M.R., Minjares, R., Castell, A., Juarez, M.D., and Rossello, C., (2011). Effects of air drying temperature on antioxidant activity and carotenoids content of carrots (Daucus carota). European drying conference.
29. Wagner, E.K., Hewlett, M.J., Bloom, D.C., and Camerini, D. (2008). Replication of some nuclear-replicating eukaryotic DNA viruses with large genomes. In: Basic virology, 3rd ed, Blackwell Publishing Ltd, pp. 331-351.
30. Wojcikowski, K., Stevenson, L., Leach, D., Wohlmuth, H., and Gobe, G. (2007). Antioxidant capacity of 55 medicinal herbs traditionally used to treat the urinary system: a comparison using a sequential threesolvent extraction process. J Alt Compl Med 13: 103–110.
31. Wojdylo, A., Oszmiansky, J., and Czemerys, R., (2007). Antioxidant activity and phenolic compounds in 32 selected herbs. Food chem. 105:940-949.
32. Xiang, Y., Pei, Y., Chang, Q., Zhizai, L., Zhe, R., Ke, Y., Sheng, X., Yingjun, Z., Chongren, Y., Dong, W.,
33. Qing, L., Kaio, K., and Yifei, W., (2011). In vitro anti-herpes simplex virus activity of 1, 2, 4, 6-tetraO-galloyl-β-D-glucose from Phyllamthus emblica L. (Euphorbiaceae). Phytotherapy Res. 25:975-982.
34. Zhang, J., B. Zhan, X. Yao, Y. Gao and J. Shong, (1995). Antiviral activity of tannin from the pericarp of Punica granatum L. against genital Herpes virus in vitro. Zhongguo Zhong Yao Za Zhi., 20: 556-576.
35. Zheng, W., and Wang, Y.(2001). Antioxidant activity and phenolic compounds in selected herbs. Food Chem. 49:5165-5170.