Prunella vulgaris L. VE Prunella grandiflora L.’DEN SAFLAŞTIRILAN ROSMARİNİK ASİTİN FARKLI TÜMÖR HÜCRELERİ ÜZERİNDEKİ SİTOTOKSİK AKTİVİTESİ

Öz

Bu çalışmada rosmarinik asit bileşiği, Prunella grandiflora L. ve P. vulgaris L. türlerinden elde edilen etanol ekstraktlarından saflaştırılmıştır. Saflaştırma işlemi sırasında elde edilen metanol fraksiyonlarının toplam fenol içeriği Folin-Ciocalteu yöntemi ile belirlenmiştir. Prunella L. türlerinden saflaştırılan rosmarinik asitin farklı kanser hücreleri üzerinde WST-1 (Roche Applied Sciences, Mannheim, Almanya) yöntemiyle sitotoksik doz çalışmaları yapılmıştır. Buna göre pankreas (PANC-1), prostat (PC-3), kolon (HT-29) ve meme (MDA-MB 436) kanserleri ile GBM (T98G) hücre hatları ve lenf dokularında 10-60ng arasında sitotoksik doz belirlenmiştir. Sitotoksik doz belirleme çalışmaları için 24 ve 48 saat inkübasyon süreleri çalışılmıştır. 48 saat inkübasyon süresi sonunda PC3 hücre hattı için 50ng ve PANC-1, HT-29, MDA-MB 436 ve T98G hücre hatları için 60ng rosmarinik asit uygulamasında antiproliferatif etki gözlenmiştir. Sağlıklı hücrelerde rosmarinik asitin sitotoksik etkisi gözlenmemiştir.

Anahtar Kelimeler

• Prunella L.,izolasyon,rosmarinic asit,antiproliferatif,sitotoksik aktivite

CYTOTOXIC ACTIVITY OF ROSMARINIC ACID ISOLATED FROM Prunella vulgaris L. AND Prunella grandiflora L. IN DIFFERENT TUMOR CELLS

Abstract

Rosmarinic acid was isolated from ethanol extractions obtained from Prunella grandiflora L. and P. vulgaris L. The total phenolic content of ethanol fractions during isolation was determined by the Folin-Ciocateu method. The cytotoxic doses of the isolated rosmarinic acid were determined by WST-1 (Roche Applied Sciences, Mannheim, Germany) cell proliferation assay. A 10-60ng cytotoxic dose was determined for pancreas (PANC-1), prostate (PC-3), colon (HT-29) and breast (MDA-MB 436) cancers and GBM (T98G) cell lines and lymphatic tissues. 24 and 48h incubation periods were applied during dose determinations. An antiproliferative effect was observed at the end of 48h incubation period with 50ng rosmaniric acid treatment in PC3 cell line and with 60ng treatment in PANC-1, HT-29, MDA-MB 436 and T98G cell lines. No cytotoxic activity of rosmarinic acid was observed in non-tumor cells

Keywords

• Prunella L.,isolation,rosmarinic acid,antiproliferative,cytotoxic activity

References

1. 1. Baricevic, D., Sosa, S., Loggia, R.D., Tubaro, A., Simonovska, B., Krasna, A. & Zupancic, A. 2001. Topical anti-inflammatory activity of Salvia officinalis L. leaves:the relevance of ursolic acid. Journal of Ethnopharmacology, 75: 125-132.
2. 2. Canturk, Z., Dikmen, M., Artagan, O., Ozarda, M.G. & Ozturk, N. Cytotoxic Effects of Resveratrol, Rutin and Rosmarinic Acid on ARH-77 Human (Multiple Myeloma) Cell Line. 2016. Natural Product Communications, 11(10): 1441-1440.
3. 3. Feng, L., Jia, X.B., Jiang, J., Zhu, M.M., Chen, Y., Tan, X.B. & Shi, F. 2010. Combination of active components enhances the efficacy of Prunella in prevention and treatment of lung cancer. Molecules, 15(11): 7893-7906.
4. 4. Hadi, N., Sefidkon, F., Shojaeiyan, A., Siler, B., Jafari, A-A., Anicic, N. & Misic, D. 2017. Phenolics’ composition in four endemic Nepeta species from Iran cultivated under experimental field conditions: The possibility of the exploitation of Nepeta germplasm. Industrial Crops and Products, 95: 475-484.
5. 5. Hollman, P.C., van Trijp, J.M., Buysman, M.N., van der Gaag, M.S., Mengelers, M.J., de Vries, J.H. & Katan, M.B. 1997. Relative bioavailability of the antioxidant flavonoid quercetin from various foods in man. FEBS Letters, 24: 418(1-2), 152-156.
6. 6. Júnior, J.T.C., Morais, S.M., Gomez, C.V., Molas, C.C., Rolon, M., Boligon, A.A., Ahtayde, M.L., Oliveira, C.D.M., Tintino, S.R. & Douglas, M.C.H. 2016. Phenolic composition and antiparasitic activity of plants from the Brazilian Northeast “Cerrado”. Saudi Journal of Biological Sciences, 23: 434–440.
7. 7. Kim, D-O. & Lee, C.Y. 2004. Comprehensive study on vitamin C equivalent antioxidant capacity (VCEAC) of various polyphenolics in scavenging a free radical and its structural relationship. Critical Reviews in Food Science and Nutrition, 44(4): 253–273.
8. 8. Petersen, M. & Simmonds, M.S.J. 2003. Rosmarinic acid. Phytochemistry, 62: 121-125.

9. 9. Singleton, V.L., Orthofer, R. & Lamuela-Raventos, R.M. 1999. Analysis of total phenols and other oxidation substrates and antioxidants by means of Folin-Ciocalteu Reagent. Methods in Enzymology, 299: 152-178.
10. 10. Şahin, S., Arı, F., Demir, C. & Ulukaya, E. 2014. Isolatıon of major phenolic compounds from the extracts of Prunella L. species grown in Turkey and their antioxidant and cytotoxic activities. Journal of Food Biochemistry, 38: 248–257.
11. 11. Tezcan, G., Tunca, B., Bekar, A., Yalçın, M., Sahin, S., Budak, F., Çeçener, G., Egeli, Ü., Demir, C., Güvenç, G., Yılmaz, G., Erkan, L.G., Malyer, H., Taşkapılıoğlu, M.Ö., Evrensel, A. & Bilir, A. 2015. Ficus carica Latex Prevents Invasion Through Induction of Let-7d Expression in GBM Cell Lines. Cellular and Molecular Neurobiology, 35(2):175-87.
12. 12. Woo, H.J., Jun Do, Y., Lee, J.Y., Woo, M.H., Yang, C.H. & Kim, Y.H. 2011. Apoptogenic activity of 2α,3α-dihydroxyurs-12-ene-28-oic acid from Prunella vulgaris var. lilacina is mediated via mitochondria-dependent activation of caspase cascade regulated by Bcl-2 in human acute leukemia Jurkat T cells. Journal of Ethnopharmacology, 135(3): 626-635.
13. 13. Yeşil-Çeliktaş, O., Sevimli, C., Bedir, E. & Vardar-Sukan, F. 2010. Inhibitory effects of rosemary extracts, carnosic acid and rosmarinic acid on the growth of various human cancer cell lines. Plant Foods Human Nutrition, 65(2): 158-163.
14. 14. Zdařilová, A., Svobodová, A., Šimánek, V. & Ulrichová, J. 2009. Prunella vulgaris extract and rosmarinic acid suppress lipopolysaccharide-induced alteration in human gingival fibroblasts. Toxicology in Vitro, 23: 386–392.