In vitro EVALUATIONS OF ANTIOXIDANT, ANTIMICROBIAL AND ANTICANCER POTENTIAL OF Phytolacca americana L. (POKEWEED) SEED EXTRACT
Year 2022,
Volume: 23 Issue: 2, 135 - 143, 15.10.2022
Elif Demirkan
,
Elif Ertürk
,
Gamze Yıldız
,
Tuba Sevgi
,
Aynur Aybey
Abstract
In this study, different parts of Phytolacca americana L. (Pokeweed) fruit from Türkiye were investigated for their antioxidant, antibacterial, antibiofilm and anticancer potentials. The radical scavenging activities, reducing power and total phenolic content were determined to appraise of the antioxidant potentials. The antibacterial and antibiofilm activities of the extracts against Enterococcus faecalis, Yersinia enterocolitica, Staphylococcus aureus, Escherichia coli, Klebsiella pneumoniae, Salmonella typhimurium were evaluated by using agar-well diffusion, minimum inhibitory concentration (MIC) and biofilm inhibitory concentration (BIC) assays. In addition of latening the onset of apoptosis depending on dose, the potential of the anti-proliferative effects was investigated on MDA-MB-231 and MCF-7 cells. The highest free radical scavenging activity and phenolic content were found in the seed extract. Seed extract showed the highest inhibition zones and significant antibacterial activity at 2.5-5 mg/mL MIC concentrations against tested bacterial strains. More significantly, seed extract was found effective on inhibition of early phase biofilm formation at 2.5-10 mg/mL. BIC concentrations against tested bacterial strains. Next, the main mechanisms of cell death of the seed extract in MDA-MB-231 and MCF-7 cells were investigated. Accordingly, when apoptosis was evaluated morphologically, late apoptosis was observed in cells that showed both Hoechst 33342 and Propidium Iodide (PI) positivity in a dose-dependent manner. This study showed that P. americana seed extract can contribute to alternative medicine studies and have potential power in pharmaceutical industry.
Supporting Institution
Bursa Uludağ University Scientific Research Project Commission
Project Number
KUAP(F)-2018/19
Thanks
The authors wish to express their hearty thanks to the late Prof. Dr. Hulusi Malyer (Bursa, TÜRKİYE) from Botanic Department, for providing the fruits of the P. americana.
References
- 1. Akgun, O., Erkisa, M. & Ari, F. 2019. Effective and new potent drug combination: histone deacetylase and Wnt/β-catenin pathway inhibitors in lung carcinoma cells. Journal of Cellular Biochemistry, 120(9): 15467-15482.
- 2. Antasionasti, I., Riyanto, S. & Rohman, A. 2017. Antioxidant activities and phenolics contents of avocado (Persea americana Mill.) peel in vitro. Journal of Medicinal Plants, 11: 55-61.
- 3. Bailly, C. & Vergoten, G. 2020. Esculentosides: Insights into the potential health benefits, mechanisms of action and molecular targets. Phytomedicine, 79: 153343.
- 4. Balogh, L. & Juhasz, J. 2008. American and Chinese pokeweed (Phytolacca americana L., Ph. esculenta van Houtte), pp. 35-46. In: Botta-Dukát, Z. & Balogh, L. (eds). The most important invasive plants in Hungary. Hungarian Academy of Sciences, Vácrátót, Hungary, 255 pp.
- 5. Blois, M. S. 1958. Antioxidant determinations by the use of a stable free radical. Nature, 181: 1199-1200.
- 6. Boo, H. O., Park, J. H., Woo, S. H. & Park, H. Y. 2015. Antimicrobial effect, antioxidant and tyrosinase inhibitory activity of the extract from different parts of Phytolacca americana L. The Korean Journal of Crop Science, 60(3): 366-373.
- 7. Bossdorf, O., Arcuri, D., Richards, C. L. & Pigliucci, M. 2010. Experimental alteration of DNA methylation affects the phenotypic plasticity of ecologically relevant traits in Arabidopsis thaliana. Evolution Ecology, 24: 541-553.
- 8. Cheesbrough, M. 2006. District laboratory practice in tropical countries Part 2. Cambridge University Press., New York, 434 pp.
- 9. Clinical and Laboratory Standards Institute (CLSI). 2005. Performance standards for antimicrobial susceptibility testing; 15th informational supplement, Clinical and Laboratory Standards Institute, Wayne, PA.
- 10. Gins, V. K., Gins, M. S., Kononkov, P. F., Pivovarov, V. F. & Kulikov, I. M. 2017. Multi-purpose use of Phytolacca with antioxidant activity. Horticulture and Viticulture, 2: 47-51.
- 11. Giovanelli, G. & Buratti, S. 2019. Comparison of polyphenolic composition and antioxidant activity of wild Italian blueberries and some cultivated varieties. Food Chemistry, 112: 903-908.
- 12. Guragac-Dereli, F. T., Onem, E., Arin, E., Ozaydin, A. G. & Muhammed, M. T. 2022. Persea americana Mill.: As a potent quorum sensing inhibitor of Pseudomonas aeruginosa PAO1 virulence. International Journal of Secondary Metabolite, 9:14-26.
- 13. Jung, C., Hong, J. Y., Bae, S. Y., Kang, S. S. & Park, H. J. 2015. Antitumor activity of Americanin isolated from the seeds of Phytolacca americana by regulating the ATM/ATR signaling pathway and the Skp2-p27 axis in human colon cancer Cells. Journal of Natural Products, 78: 2983-93.
- 14. Karker, M., Falleh, H., Msaada, K., Smaoui, A. & Abdelly, C. 2016. Antioxidant, anti-inflammatory and anticancer activities of the medicanal halophyte Reumuria vermiculata. Experimental and Clinical Sciences, 15: 297-307.
- 15. Liberato, S. F. D. S., Vegian, M. R. C., Hasna, A. A., Alvarenga, J. D., Santos, J. G., Tini, I. R. P., Amêndola, I., Junqueira, J. P. & Oliveira less, L. D. 2021. Antibiofilm action of Persea americana glycolic extract over Acinetobacter baumannii and absence of toxicity in Galleria mellonella. Journal of Complementary and Integrative Medicine, Article number: 000010151520210051.
- 16. Maness, L., Goktepe, I., Hardy, B., Yu, J. & Ahmedna, M. 2012. Antiproliferative and apoptotic effects of Phytolacca americana extracts and their fractions on breast and colon cancer cells. Research Journal of Medicinal Plants, 6: 17-26.
- 17. Maness, L., Goktepe, I., Chen, H., Ahmedna, M. & Sang, S. 2014. Impact of Phytolacca americana extracts on gene expression of colon cancer cells. Phytotheraphy Research, 28: 219-223.
- 18. Mchedlishvili, N. I., Omiadze, N. T., Abutidze, M. O., Rodriguez-Lope, J. N. & Sadunishvili, T. A. 2014. Investigation of phenolic content, antioxidant and antimicrobial activities of natural food red colorant from Phytolacca americana L. fruits. Annals of Agricultural Sciences, 12: 71-75.
- 19. Meesilp, N. & Mesil, N. 2019. Effect of microbial sanitizers for reducing biofilm formation of Staphylococcus aureus and Pseudomonas aeruginosa on stainless steel by cultivation with UHT milk. Food Science and Biotechnology, 28: 289-296.
- 20. Merghni, A., Noumi, E., Hadded, O., Dridi, N. & Panwar, H. 2018. Assessment of the antibiofilm and antiquorum sensing activities of Eucalyptus globulus essential oil and its main component 1, 8-cineole against methicillin-resistant Staphylococcus aureus strains. Microbial Pathogenesis, 118: 74-80.
- 21. Miliauskas, G., Venskutonis, P.R. & Van Beek, T.A. 2004. Screening of radical scavenging activity of some medicinal and aromatic plant extracts. Food Chemistry, 85: 231-237.
- 22. Nabavi, S. M., Ebrahimzadeh, M. A., Nabavi, S. F. & Bahramian, F. 2009. In vitro antioxidant activity of Phytolacca americana berries. Pharmacology, 1: 81-88.
- 23. Nithya, C., Pandian, S. K. 2010. The in vitro antibiofilm activity of selected marine bacterial culture supernatants against Vibrio spp. Archives of Microbiology, 192: 843-854.
- 24. Nunes, P. X., Silva, S. F., Guedes, R. J. & Almeida, S. 2012. Biological oxidations and antioxidant activity of natural products. pp 1-20. In: Rao, V. (eds). Phytochemicals as Nutraceuticals - Global Approaches to Their Role in Nutrition and Health. IntechOpen, Newyork, 290 pp.
- 25. Oyaizu, M. 1986. Studies on product of browning reaction prepared from glucose amine. The Japanese Journal of Nutrition and Dietetics, 44: 307-315.
- 26. Patra, J. K., Kim, E. S., Oh, K., Kim, H. J. & Kim, Y. 2014. Antibacterial effect of crude extract and metabolites of Phytolacca americana on pathogens responsible for periodontal inflammatory diseases and dental caries. BMC Complementary Medicine and Therapies, 14: 343-350.
- 27. Re, R., Pellegrini, N., Proteggente, A., Pannala, A., Yang, M. & Rice-Evans, C. 1999. Antioxidant activity applying an improved ABTS radical cation decolorization assay. Free Radical Biology and Medicine, 26: 1231-1237.
- 28. Sarabhai, S., Sharma, P. & Capalash, N. 2013. Ellagic acid derivatives from terminalia chebula retz. Downregulate the expression of quorum sensing genes to attenuate Pseudomonas aeruginosa PAO1 virulence. PLoS ONE, 8: e53441.
- 29. Solís-Salas, M. L., Sierra-Rivera, C. A., Cobos-Puc, L. E., Ascacio-Valdés, J. A. & Silva-Belmares, S. Y. 2021. Antibacterial Potential by Rupture Membrane and Antioxidant Capacity of Purified Phenolic Fractions of Persea americana Leaf Extract. Antibiotics, 10:508.
- 30. Singleton, V. L. & Rossi, J. A. 1965. Colorimetry of total phenolics with phosphomolybdic phosphotungstic acid reagents. American Journal of Enology and Viticulture, 16: 144-158.
- 31. Wannes, W. A., Mhamdi, B., Sriti, J., Jemia, M. B., Ouchikh, O., Hamdaoui, G., Kchouk, M. E. & Marzouk, B. 2010. Antioxidant activities of the essential oil and methanol extracts from myrtle (Myrtus communis var. italica L.) leaf, stem and flower. Food and Chemical Toxicology, 48: 1362-1370.
- 32. Wilkins, M., Hall-Stoodley, L., Allan, R. N., Faust, S. N. 2014. New approaches to the treatment of biofilm-related infections. The Journal of Infectious Diseases, 69: 47-52.
- 33. Vichai, V. & Kirtikara, K. 2006. Sulforhodamine B colorimetric assay forcytotoxicity screening. Nature Protocols, 1: 1112-1116.
- 34. Zheleva-Dimitrova, D. Z. 2019. Antioxidant and acetylcholinesterase inhibition properties of Amorpha fruticosa L. and Phytolacca americana L. Pharmacognosy Magazine. 9: 109-113.
Year 2022,
Volume: 23 Issue: 2, 135 - 143, 15.10.2022
Elif Demirkan
,
Elif Ertürk
,
Gamze Yıldız
,
Tuba Sevgi
,
Aynur Aybey
Abstract
Bu çalışmada, Türkiye'de yetişen Phytolacca americana L. (Pokeweed) meyvesinin farklı kısımlarının antioksidan, antibakteriyel, antibiyofilm ve antikanser potansiyelleri araştırılmıştır. Antioksidan potansiyellerini değerlendirmek için radikal süpürücü aktiviteleri, indirgeme aktivitesi ve toplam fenolik içerik belirlendi. Ekstraktın Enterococcus faecalis, Yersinia enterocolitica, Staphylococcus aureus, Escherichia coli, Klebsiella pneumoniae, Salmonella typhimurium'a karşı antibakteriyel ve antibiyofilm aktiviteleri, agar-kuyu difüzyon, minimum inhibitör konsantrasyon (MIC) ve biyofilm inhibitör konsantrasyon (BIC) testleri yapılarak değerlendirildi. Doza bağlı olarak apoptoz başlangıcını geciktirmenin yanı sıra, MDA-MB-231 ve MCF-7 hücreleri üzerinde anti-proliferatif etkilerin potansiyeli araştırıldı. En yüksek serbest radikal süpürücü aktivite ve fenolik içerik, çekirdek ekstraktında bulundu. Çekirdek ekstraktı en yüksek inhibisyon zonu ve test edilen bakteri suşlarına karşı da 2.5-5 mg/mL MIC konsantrasyonlarında önemli bir antibakteriyel aktivite gösterdi. Daha da önemlisi, çekirdek ekstraktının test edilen bakteri suşlarına karşı 2.5-10 mg/mL BIC konsantrasyonlarında erken faz biyofilm oluşumunun inhibisyonunda etkili olduğu bulunmuştur. Ek olarak, P. americana meyvesinin diğer kısımlarına kıyasla çekirdek ekstraktı, 48 ve 72 saat boyunca farklı konsantrasyonlarda hücreler üzerinde anlamlı anti-büyüme etkileri sergilemiştir. Daha sonra, MDA-MB-231 ve MCF-7 hücrelerinde çekirdek ekstraktının hücre ölümünün ana mekanizmaları araştırıldı. Buna göre apoptoz morfolojik olarak değerlendirildiğinde, doza bağlı olarak hem Hoechst 33342 hem de Propidium Iodide (PI) pozitifliği gösteren hücrelerde geç apoptoz gözlendi. Bu çalışma P. americana çekirdek ekstraktının alternatif tıp çalışmalarına katkıda bulunabileceğini ve ilaç endüstrisinde potansiyel bir güce sahip olabileceğini göstermiştir.
Project Number
KUAP(F)-2018/19
References
- 1. Akgun, O., Erkisa, M. & Ari, F. 2019. Effective and new potent drug combination: histone deacetylase and Wnt/β-catenin pathway inhibitors in lung carcinoma cells. Journal of Cellular Biochemistry, 120(9): 15467-15482.
- 2. Antasionasti, I., Riyanto, S. & Rohman, A. 2017. Antioxidant activities and phenolics contents of avocado (Persea americana Mill.) peel in vitro. Journal of Medicinal Plants, 11: 55-61.
- 3. Bailly, C. & Vergoten, G. 2020. Esculentosides: Insights into the potential health benefits, mechanisms of action and molecular targets. Phytomedicine, 79: 153343.
- 4. Balogh, L. & Juhasz, J. 2008. American and Chinese pokeweed (Phytolacca americana L., Ph. esculenta van Houtte), pp. 35-46. In: Botta-Dukát, Z. & Balogh, L. (eds). The most important invasive plants in Hungary. Hungarian Academy of Sciences, Vácrátót, Hungary, 255 pp.
- 5. Blois, M. S. 1958. Antioxidant determinations by the use of a stable free radical. Nature, 181: 1199-1200.
- 6. Boo, H. O., Park, J. H., Woo, S. H. & Park, H. Y. 2015. Antimicrobial effect, antioxidant and tyrosinase inhibitory activity of the extract from different parts of Phytolacca americana L. The Korean Journal of Crop Science, 60(3): 366-373.
- 7. Bossdorf, O., Arcuri, D., Richards, C. L. & Pigliucci, M. 2010. Experimental alteration of DNA methylation affects the phenotypic plasticity of ecologically relevant traits in Arabidopsis thaliana. Evolution Ecology, 24: 541-553.
- 8. Cheesbrough, M. 2006. District laboratory practice in tropical countries Part 2. Cambridge University Press., New York, 434 pp.
- 9. Clinical and Laboratory Standards Institute (CLSI). 2005. Performance standards for antimicrobial susceptibility testing; 15th informational supplement, Clinical and Laboratory Standards Institute, Wayne, PA.
- 10. Gins, V. K., Gins, M. S., Kononkov, P. F., Pivovarov, V. F. & Kulikov, I. M. 2017. Multi-purpose use of Phytolacca with antioxidant activity. Horticulture and Viticulture, 2: 47-51.
- 11. Giovanelli, G. & Buratti, S. 2019. Comparison of polyphenolic composition and antioxidant activity of wild Italian blueberries and some cultivated varieties. Food Chemistry, 112: 903-908.
- 12. Guragac-Dereli, F. T., Onem, E., Arin, E., Ozaydin, A. G. & Muhammed, M. T. 2022. Persea americana Mill.: As a potent quorum sensing inhibitor of Pseudomonas aeruginosa PAO1 virulence. International Journal of Secondary Metabolite, 9:14-26.
- 13. Jung, C., Hong, J. Y., Bae, S. Y., Kang, S. S. & Park, H. J. 2015. Antitumor activity of Americanin isolated from the seeds of Phytolacca americana by regulating the ATM/ATR signaling pathway and the Skp2-p27 axis in human colon cancer Cells. Journal of Natural Products, 78: 2983-93.
- 14. Karker, M., Falleh, H., Msaada, K., Smaoui, A. & Abdelly, C. 2016. Antioxidant, anti-inflammatory and anticancer activities of the medicanal halophyte Reumuria vermiculata. Experimental and Clinical Sciences, 15: 297-307.
- 15. Liberato, S. F. D. S., Vegian, M. R. C., Hasna, A. A., Alvarenga, J. D., Santos, J. G., Tini, I. R. P., Amêndola, I., Junqueira, J. P. & Oliveira less, L. D. 2021. Antibiofilm action of Persea americana glycolic extract over Acinetobacter baumannii and absence of toxicity in Galleria mellonella. Journal of Complementary and Integrative Medicine, Article number: 000010151520210051.
- 16. Maness, L., Goktepe, I., Hardy, B., Yu, J. & Ahmedna, M. 2012. Antiproliferative and apoptotic effects of Phytolacca americana extracts and their fractions on breast and colon cancer cells. Research Journal of Medicinal Plants, 6: 17-26.
- 17. Maness, L., Goktepe, I., Chen, H., Ahmedna, M. & Sang, S. 2014. Impact of Phytolacca americana extracts on gene expression of colon cancer cells. Phytotheraphy Research, 28: 219-223.
- 18. Mchedlishvili, N. I., Omiadze, N. T., Abutidze, M. O., Rodriguez-Lope, J. N. & Sadunishvili, T. A. 2014. Investigation of phenolic content, antioxidant and antimicrobial activities of natural food red colorant from Phytolacca americana L. fruits. Annals of Agricultural Sciences, 12: 71-75.
- 19. Meesilp, N. & Mesil, N. 2019. Effect of microbial sanitizers for reducing biofilm formation of Staphylococcus aureus and Pseudomonas aeruginosa on stainless steel by cultivation with UHT milk. Food Science and Biotechnology, 28: 289-296.
- 20. Merghni, A., Noumi, E., Hadded, O., Dridi, N. & Panwar, H. 2018. Assessment of the antibiofilm and antiquorum sensing activities of Eucalyptus globulus essential oil and its main component 1, 8-cineole against methicillin-resistant Staphylococcus aureus strains. Microbial Pathogenesis, 118: 74-80.
- 21. Miliauskas, G., Venskutonis, P.R. & Van Beek, T.A. 2004. Screening of radical scavenging activity of some medicinal and aromatic plant extracts. Food Chemistry, 85: 231-237.
- 22. Nabavi, S. M., Ebrahimzadeh, M. A., Nabavi, S. F. & Bahramian, F. 2009. In vitro antioxidant activity of Phytolacca americana berries. Pharmacology, 1: 81-88.
- 23. Nithya, C., Pandian, S. K. 2010. The in vitro antibiofilm activity of selected marine bacterial culture supernatants against Vibrio spp. Archives of Microbiology, 192: 843-854.
- 24. Nunes, P. X., Silva, S. F., Guedes, R. J. & Almeida, S. 2012. Biological oxidations and antioxidant activity of natural products. pp 1-20. In: Rao, V. (eds). Phytochemicals as Nutraceuticals - Global Approaches to Their Role in Nutrition and Health. IntechOpen, Newyork, 290 pp.
- 25. Oyaizu, M. 1986. Studies on product of browning reaction prepared from glucose amine. The Japanese Journal of Nutrition and Dietetics, 44: 307-315.
- 26. Patra, J. K., Kim, E. S., Oh, K., Kim, H. J. & Kim, Y. 2014. Antibacterial effect of crude extract and metabolites of Phytolacca americana on pathogens responsible for periodontal inflammatory diseases and dental caries. BMC Complementary Medicine and Therapies, 14: 343-350.
- 27. Re, R., Pellegrini, N., Proteggente, A., Pannala, A., Yang, M. & Rice-Evans, C. 1999. Antioxidant activity applying an improved ABTS radical cation decolorization assay. Free Radical Biology and Medicine, 26: 1231-1237.
- 28. Sarabhai, S., Sharma, P. & Capalash, N. 2013. Ellagic acid derivatives from terminalia chebula retz. Downregulate the expression of quorum sensing genes to attenuate Pseudomonas aeruginosa PAO1 virulence. PLoS ONE, 8: e53441.
- 29. Solís-Salas, M. L., Sierra-Rivera, C. A., Cobos-Puc, L. E., Ascacio-Valdés, J. A. & Silva-Belmares, S. Y. 2021. Antibacterial Potential by Rupture Membrane and Antioxidant Capacity of Purified Phenolic Fractions of Persea americana Leaf Extract. Antibiotics, 10:508.
- 30. Singleton, V. L. & Rossi, J. A. 1965. Colorimetry of total phenolics with phosphomolybdic phosphotungstic acid reagents. American Journal of Enology and Viticulture, 16: 144-158.
- 31. Wannes, W. A., Mhamdi, B., Sriti, J., Jemia, M. B., Ouchikh, O., Hamdaoui, G., Kchouk, M. E. & Marzouk, B. 2010. Antioxidant activities of the essential oil and methanol extracts from myrtle (Myrtus communis var. italica L.) leaf, stem and flower. Food and Chemical Toxicology, 48: 1362-1370.
- 32. Wilkins, M., Hall-Stoodley, L., Allan, R. N., Faust, S. N. 2014. New approaches to the treatment of biofilm-related infections. The Journal of Infectious Diseases, 69: 47-52.
- 33. Vichai, V. & Kirtikara, K. 2006. Sulforhodamine B colorimetric assay forcytotoxicity screening. Nature Protocols, 1: 1112-1116.
- 34. Zheleva-Dimitrova, D. Z. 2019. Antioxidant and acetylcholinesterase inhibition properties of Amorpha fruticosa L. and Phytolacca americana L. Pharmacognosy Magazine. 9: 109-113.