Öz

Doğal kaynaklardan yeni ve etkili antiviral etkenler bulmak amacıyla yapılan bu çalışmada, Ballota glandulosissima Hub.-Mor. & Patzak’dan elde edilen metanol ve su ekstraktlarının antiviral özellikleri Respiratuvar Sinsityal Virus (RSV)’una karşı kolorimetrik XTT testi ile değerlendirilmiştir. Virüsün neden olduğu sitopatik etkilere karşı %50 koruma sağlaması için gerekli konsantrasyon EC₅₀ olarak tanımlanmış, CC₅₀ (%50 Sitotoksik Konsantrasyon)’nin EC₅₀’ye oranından da seçicilik indeksi (SI) belirlenmiştir. Araştırma sonucunda, B. glandulosissima’nın hem metanol (EC₅₀ = 12.45 µg/mL; SI = 24.84) hem de su ekstraktının (EC₅₀ = 19.12 µg/mL; SI = 24.59) RSV’ye karşı; pozitif kontrol olarak kullanılan ribavirin (EC₅₀ = 3.25 µg/mL, SI = 34.89)’e göre önemli sayılabilecek oranda anti-RSV aktiviteye sahip olduğu tespit edilmiştir. Sonuç olarak, B. glandulosissima ekstraktlarının, RSV’ye karşı klinikte kullanılan ilaçlara karşı bir alternatif olarak geliştirilebilmesi için; daha ileri çalışmalara layık olduğunu söyleyebiliriz. Bu çalışma, B. glandulosissima’nın anti-RSV aktivitesine yönelik ilk rapordur.

EVALUATION OF THE ANTIVIRAL ACTIVITY OF Ballota glandulosissima Hub.-Mor. & Patzak EXTRACTS AGAINST RESPIRATORY SYNCYTIAL VIRUS (RSV)

Abstract

In order to find new and effective antiviral agents from natural sources, the antiviral properties of methanol and aqueous extracts obtained from Ballota glandulosissima Hub.-Mor. & Patzak were evaluated by Colorimetric XTT test against Respiratory syncytial virus (RSV). The concentration required to provide 50% protection against cytopathic effects caused by the virus was defined as EC₅₀, the selectivity index (SI) was determined from the ratio of CC₅₀ (50 % Cytotoxic concentration) to EC₅₀. The results showed that both the methanol (EC₅₀ = 12.45 µg/mL; SI = 24.84) and aqueous extracts (EC₅₀ = 19.12 µg/mL; SI = 24.59) of B. glandulosissima had almost the same strong anti-RSV activity as well as ribavirin, used as a positive control against RSV (EC₅₀ = 3.25 µg/mL, SI = 34.89). In conclusion, we can say that B. glandulosissima extracts are worthy of further studies in order to develop an alternative to the drugs used in clinical practice against RSV. This is the first report on the anti-RSV activity of B. glandulosissima. 

Keywords

• Ballota glandulosissima,methanol and aqueous extracts,antiviral activity,respiratory syncytial virus

References

1. 1. Andrighetti-Fröhmer, C.R., Antonio, R.V., Creczynski-Pasa, T.B., Barardi, C.R.M. & Simões, C.M.O. 2003. Cytotoxicity and potential antiviral evaluation of violacein produced by Chromobacterium violaceum. Memórias do Instituto Oswaldo Cruz, 98: 843-848.
2. 2. Arita, M., Philipov, S. & Galabov, A.S. 2015. Phosphatidylinositol 4-kinase III beta is the target of oxoglaucine and pachypodol (Ro 09-0179) for their anti-poliovirus activities, and is located at upstream of the target step of brefeldin A. Microbiology and Immunulogy, 59(6): 338-347.
3. 3. Baytop, T. 1984. Türkiye’de Bitkilerle Tedavi. İstanbul Üniversitesi Yayınları, İstanbul, 371 pp.
4. 4. Chattopadhyay, D., Chawla-Sarkar, M., Chatterjee, T., Dey, R.S., Bag, P., Chakraborti, S. & Khan, M.T.H. 2009. Recent advancements for the evaluation of antiviral activities of natural products. New Biotechnology, 25: 347-368.
5. 5. Chiang, L.C., Chiang, W., Chang, M.Y., Ng, L.T. & Lin, C.C. 2002. Antiviral activity of Plantago major extracts and related compounds in vitro. Antiviral Research, 55: 53-62.
6. 6. Chin, J., Magoffin, R.L., Shearer, L.A., Schieble, J.H. & Lennette, EH. 1969. Field evaluation of a respiratory syncytial virus vaccine and a trivalent parainfluenza virus vaccine in a paediatric population. American Journal of Epidemiology, 89: 449-463.
7. 7. Çitoğlu, G., Tanker, M. & Sever, B. 1999. Note flavonoid aglycones from B. saxatilis subsp. saxatilis. Pharmaceutical Biology, 37: 158-160.
8. 8. Çitoğlu, G., Tanker, M., Sever, B., Englert, J., Anton, R. & Altanlar, N. 1998. Antibacterial activities of diterpenoids isolated from B. saxatilis subsp. saxatilis. Planta Medica, 64: 484-485.

9. 9. Cos, P., Vlietinck, A.J., Vanden Berghe, D. & Maes, L. 2006. Anti-infective potential of natural products: how to develop a stronger in vitro ‘proof-of concept’. Journal of Ethnopharmacology, 106: 290-302.
10. 10. Davis, P.H. 1982. Flora of Turkey and the East Aegean Islands. Edinburgh University Press, Edinburg, volume 7, 156-160 pp.
11. 11. Dubovi, E.J., Geratz, J.D., Shaver, S.R. & Tidwell, R.R. 1981. Inhibition of respiratory syncytial virus-host cell interactions by mono and diamidines. Antimicrobial Agents and Chemotherapy, 19: 649-656.
12. 12. Englund, J.A., Sullivan, C.J., Jordan, M.C., Dehner, L.P., Verecellotti, G.M. & Balfour, F. 1988. Respiratory syncytial virus infection in immunocompromised adults. Annals of Internal Medicine, 109: 203-208.
13. 13. Erdoğan-Orhan, I., Sever-Yılmaz, B., Altun, M.L. & Saltan Çitoğlu, G. 2010. Radical quenching activity, ferric reduction antioxidant power, and ferrous ion-chelating capacity of Ballota species and their total phenol and flavonoid contents. Journal of Medicinal Food, 13(6): 1537-1543.
14. 14. Eryaşar, P. & Tuzlacı, E. 1998. Gönen (Balıkesir) halk hekimliğinde kullanılan bitkiler, 95-102. Paper presented at XII. Bitkisel İlaç Hammaddeleri Toplantısı, 20-22 May, Hacettepe, Ankara-Turkey.
15. 15. Eskiocak, U., İşeri, O.D., Kars, M.D., Biçer, A. & Gündüz, U. 2008. Effect of doxorubicin on telomerase activity and apoptotic gene expression in doxorubicin-resistant and -sensitive MCF-7 cells: an experimental study. Chemotherapy, 54: 209-216.
16. 16. Fraser, A.W. & Lewis, J.R. 1973. Two flavonols from Euodia glabra. Phytochemistry 12: 1787-1789.
17. 17. Garnier, G., Debraux, G. & Bezanger-Beauquesne, L. 1961. Ressources Medicinales de la Flöre Française. Vigot Freres Editeurs, Paris, 1156-1158 pp.
18. 18. Greuter, W. & Raus, T. 1998. Med-checklist Notulae, 17. Willdenowia, 28: 163-174.
19. 19. Güner, A., Aslan, S., Ekim, T., Vural, M & Babaç, M.T. (edlr.) (2012). Türkiye Bitkileri Listesi (Damarlı Bitkiler). Nezahat Gökyiğit Botanik Bahçesi ve Flora Araştırmaları Derneği Yayını.
20. 20. Harrington, R.D., Hooton, T.M., Hackman, R.C., Storch, G.A., Osborne, B., Gleaves, C.A., Benson, A. & Meyers, JD. 1992. An outbreak of respiratory syncytial virus in bone marrow transplant centre. Journal of Infectious Diseases, 165: 987-993.
21. 21. Hruska, J.F., Morrow, P.E., Suffin, S.C. & Douglas, R.G. 1982. In vivo inhibition of respiratory syncytial virus by ribavirin. Antimicrobial Agents and Chemotherapy, 21: 125-130.
22. 22. Ishitsuka, H., Ninomiya, Y. & Suhara, Y. 1986. Molecular basis of drug resistance to new antirhinovirus agents. The Journal of Antimicrobial Chemotherapy, 18 Suppl B: 11-18.
23. 23. Kaerber, G. 1964. Diagnostic procedures for virus and rickettsial disease. Public Health Association, 3: 48-50.
24. 24. Kim, H.W., Canchola, J.G., Brandt, C.D., Pyles, G., Chanock, R.M., Jensen, K. & Parrott, R.H. 1969. Respiratory syncytial virus disease in infants despite prior administration of antigenic inactivated vaccine. American Journal of Epidemiology, 89: 422-434.
25. 25. Kimura, K., Mori, S., Tomita, K., Ohno, K., Takahashi, K., Shigeta, S. & Terada, M. 2000. Antiviral activity of NMSO3 against respiratory syncytial virus infection in vitro and in vivo. Antiviral Research 47: 41-51.
26. 26. Kneyber, M.C.J., Mou, H.A. & Groot, R.D. 2000. Treatment and prevention of respiratory syncytial virus infection. European Journal of Paediatrics, 159: 339-411.
27. 27. Lewinsohn, D.M., Bowden, R.A., Mattson, D. & Crawford, S.W. 1996. Phase I study of intravenous ribavirin treatment of respiratory syncytial virus pneumonia after marrow transplantation. Antimicrobial Agents and Chemotherapy, 40: 2555-2557.
28. 28. Ma, S.C., Du, J., But, P.P.H., Deng, X.L., Zhang, Y.W., Ooi, V.E.C., Xu, H.X., Lee, S.H.S. & Lee, S.F. 2002. Antiviral Chinese medicinal herbs against respiratory syncytial virus. Journal of Ethnopharmacology, 79: 205-211.
29. 29. MacDonald, NE., Hall, CB., Suffin, SC., Alexson, C., Harris, PJ. & Manning, JA. 1982. Respiratory syncytial virus infection in infants with congenital heart disease. New England of Journal Medicine, 307: 397-400.
30. 30. Meriçli, A.H., Meriçli, F. & Tuzlacı, E. 1988. Flavonoids of Ballota acetabulosa. Acta Pharmaceutica Turcica, 30: 143-144.
31. 31. Miyazawa, M., Okuno, Y., Nakamura, S. & Kosaka, H. 2000. Antimutagenic activity of flavonoids from Pogostemon cablin. Journal of Agricultural and Food Chemistry, 48(3): 642-647.
32. 32. Ninomiya, Y., Aoyama, M., Umeda, I., Suhara, Y. & Ishitsuka, H. 1985. Comparative studies on the modes of action of the Antirhinovirus agents Ro 09-0410, Ro 09-0179, RMI-15,731, 4’,6-dichloroflavan and enviroxime. Antimicrobial Agents and Chemotherapy, 27(4): 595-599.
33. 33. Orabi, K.Y., Mossa, J.S., Muhammed, I., Alloush, M.H., Galal, A.M., El-Feraly, F.S. & McPhail, E.T. 2000. New eudesmane sesquiterpene from Plectranthus cylindraceus. Journal of Natural Products, 63: 1665-1668.
34. 34. Özbek, H., Saltan Çitoğlu, G., Dülger, H., Uğraş, S. & Sever, B. 2004. Hepatoprotective and anti-inflammatory activities of Ballota glandulosissima. Journal of Ethnopharmacology, 95: 143-149.
35. 35. Patzak, V.A. 1958. Revision der gattung Ballota section Acanthoprasium und section Beringeria. Annalen des Naturhistorischen Museums in Wien, 62: 57-86.
36. 36. Patzak, V.A. 1960. Zwei nene Ballota-arten aus der Türkei nebst einem nachtrag zur gattung Ballota. Annalen des Naturhistorischen Museums in Wien, 64: 42-56.
37. 37. Pérez, L. & Carrasco, L. 1992. Lack of direct correlation between p220 cleavage and the Shut-off of host translation after Poliovirus infection. Virology, 189: 178-186.
38. 38. Rigano, D., Marrelli, M., Formisano, C., Menichini, F., Senatore, F., Bruno, M. & Conforti, F. 2016. Phytochemical profile of three Ballota species essential oils and evaluation of the effects on human cancer cells. Natural Product Research, 17: 1-9.
39. 39. Rukunga, G. & Simons, A.J. (2006). The potential of plants as a source of antimalarial agents, A review, 72pp, Africa herbal antimalarial meeting, CDE and ICRAF, World Agroforestry Centre, Nairobi, Kenya.
40. 40. Sakakibara, M., Difeo, D., Nakatani, J.N., Timmermann, B. & Mabry, T.S. 1976. Flavonoid methyl ethers on the external leaf surface of Larrea tridentata and Larrea divaricata. Phytochemistry, 15: 727-731.
41. 41. Saltan Çitoğlu G. & Sever, B. 2002. Ballota glandulosissima Hub.-Mor & Patzak ve Ballota inaequidens Hub.-Mor & Patzak’dan elde edilen flavonoid aglikonlari, 172-181, 14. Bitkisel İlaç Hammaddeleri Toplantısı, 29-31 Mayıs, Eskişehir, Türkiye.
42. 42. Saltan Çitoğlu, G., Çoban, T., Sever, B. & İşcan, M. 2004a. Antioxidant properties of Ballota species growing in Turkey. Journal of Ethnopharmacology, 92: 275-280.
43. 43. Saltan Çitoğlu, G., Özbek, H. & Sever, B. 2004b. Antinociceptive activity of Ballota glandulosissima Hub.-Mor. & Patzak. Eastern Journal of Medicine, 9(2): 55-58.
44. 44. Saltan Çitoğlu, G., Sever Yılmaz, B. & Altanlar, N. 2003a. Antimicrobial activity of Ballota species growing in Turkey. Ankara Üniversitesi Eczacılık Fakültesi Dergisi, 32(2): 93-97.
45. 45. Saltan Çitoğlu, G., Sever, B., Antus, S., Baitz-Gacs, E. & Altanlar, N. 2003b. Antifungal flavonoids from Ballota glandulosissima. Pharmaceutical Biology, 41: 483-486.
46. 46. Saltan Çitoğlu, G., Sever, B., Antus, S., Baitz-Gács, E. & Altanlar, N. 2004c. Antifungal diterpenoids and flavonoids from Ballota inaequidens. Pharmaceutical Biology, 42(8): 659-663.
47. 47. Sever Yılmaz, B., Altanlar, N. & Saltan Çitoğlu, G. 2005. Antilisterial activity of Ballota species growing in Turkey. Ankara Üniversitesi Eczacılık Fakültesi Dergisi, 34(3): 155-164.
48. 48. Sever Yılmaz, B., Özbek, H. & Saltan Çitoğlu, G. 2006. Antinociceptive and anti-inflammatory activities of Ballota inaequidens. Pharmaceutical Biology, 44(8): 636-641.
49. 49. Smith, D.W., Feankel, L.R., Mathers, L.H., Tang, A.T.S., Ariagno, R.L. & Prober, C.G. 1991. A controlled trial of aerosolized ribavirin in infants receiving mechanical ventilation for severe respiratory syncytial virus infections. New England Journal of Medicine, 325: 24-29.
50. 50. Tolon Fenercioğlu, E. & Tuzlacı, E. 1998. Şile (İstanbul) halk hekimliğinde kullanılan bitkiler, 95-102, Paper presented at XII. Bitkisel Ilaç Hammaddeleri Toplantısı, 20-22 Mayıs, Hacettepe, Ankara-Türkiye.
51. 51. Treanor, J. & Falsey, A. 1999. Respiratory viral infections in the elderly. Antiviral Research, 44: 79-102.
52. 52. Tuzlacı, E. & Tolon, E. 2000. Folk medicinal plants of Şile (Istanbul, Turkey). Fitoterapia, 71: 673-685.
53. 53. Valesi, A., Rodriguez, E., Velde, G.V. & Mabry, T.J. 1972. Methylated flavonols in Larrea cuneifolia. Phytochemistry, 11: 2821-2826.
54. 54. Vanden Berghe, D.A.R., Haemers, A. & Vlietinck, A.J. 1993. Antiviral agents from higher plants and an example of structural-activity relationships of 3-methoxyflavones. In: Colegate, S.M., Molyneux, R.J. (Eds.), Bioactive Natural Products, CRC Press, Boca Raton, 405-440 pp.
55. 55. Wollenweber, E., Schober, I., Clark, W.D. & Yatskievych, G. 1985. Flavonoid aglycones from leaf resins of two species Heterotheca (Compositae). Phytochemistry, 24(9): 2129-2131.
56. 56. Wyde, P.R., Moore-Poveda, D.K., Hara, B., Ding, W.D., Mitsner, B. & Gilbert, B.E. 1998. CL387626 exhibits marked and unusual antiviral activity against respiratory syncytial virus in tissue culture and in cotton rats. Antiviral Research, 38: 31-42.
57. 57. Yeşilada, E., Honda, G., Sezik, E., Tabata, M., Fujita, T., Tanaka, T., Takeda, Y. & Takaishi, Y. 1995. Traditional medicine in Turkey. V. folk medicine in the inner Taurus Mountains. Journal of Ethnopharmacology, 46: 133-152.
58. 58. Yeşilada, E., Honda, G., Sezik, E., Tabata, M., Goto, K. & Ikeshiro, Y. 1993. Traditional medicine in Turkey IV. Folk medicine in the Mediterranean subdivision. Journal of Ethnopharmacology, 39: 31-38.