Tülay AŞKIN ÇELİK, Özlem Sultan ASLANTÜRK

Carpobrotus acınaiformis L. Metanol Ekstresinin Fitokimyasal Taraması ve Ekstrenin İnsan Metastatik Meme Kanseri (MCF-7) ve İnsan Kolon Kanseri (Caco-2) Hücreleri Üzerindeki Sitotoksik ve Apoptotik Etkisinin Araştırılması

Bu çalışmanın amacı, C. acinaciformis L. (makas otu) bitkisinin toprak üstü kısımlarından elde edilen metanol ekstresinde (ME) fitokimyasal madde taramasının yapılması, ekstrenin toplam fenolik madde miktarı, antioksidan aktivitesi ve in vitro sitotoksik ve apoptotik etkilerinin araştırılmasıdır. Ekstrede bulunan fitokimyasallar standart metod ve HPLC analizi kullanılarak belirlenmiştir. C. acinaciformis ME, antioksidan aktivitesi (DPPH radikalini süpürme, H2O2 süpürme and metal şelatlama assay), sitotoksik etki (MCF-7 ve Caco-2 hücrelerinde MTT assay) ve apoptotik etki (DNA difüzyon assay) bakımından test edilmiştir. Yapılan kalitatif fitokimyasal tarama test sonuçlarına göre C. acinaciformiş ME’de fenoller, taninler, flavonoidler ve antrakinon’ların bulunduğu, alkaloid ve saponinin ise bulunmadığı belirlenmiştir. Ekstrenin toplam fenolik madde miktarı 61.26 ± 0,110 mg GAE/g ekstre olarak hesaplanmıştır. ME’nin denenen bütün konsantrasyonlarda (100 μg/ml, 250 μg/ml, 500 μg/ml ve 1000 μg/ml) yüksek seviyede DPPH radikalini süpürme aktivitesine sahip olduğu H2O2 süpürme aktivitesinin çok düşük olduğu, metal şelatlama aktivitesinin ise bulunmadığı belirlenmiştir. ME, MCF-7 hücreleri üzerinde oldukça yüksek oranda sitotoksik etki göstermiş (p<0.05), Caco-2 hücreleri üzerindeki sitotoksik etkisi ise daha düşük oranda olmuştur. ME’nin MCF-7 ve Caco-2 hücreleri üzerinde oluşturduğu apoptotik etki de sitotoksik etkiye benzerlik göstemiştir. Metanol ekstresinin MCF-7 hücreleri üzerindeki apoptotik etkisi, Caco-2 hücrelerine nazaran daha fazla olmuş ve aradaki farkın istatistiki açıdan da önemli olduğu bulunmuştur (p<0.05).

Anahtar Kelimeler:

Carpobrotus acinaciformis, antioksidan aktivite, sitotoksik etki, apoptotik etki

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REFERANS LİSTESİ
[1] Visioli, F., Borsani, L. ve Gali, C. (2000). Diet and prevention of coronary heart disease: the potential role of phytochemicals. Cardiovasc. Res., 47, 149-425.
[2] Güney, O., Canbilen, A., Konak, A. ve Acar, O. (2003). The effects of folic acid in the prevention of neural tube development defects caused by phenytoin in early chick embryos. Spine, 28(5), 442-445.
[3] Farr, D. R. (1997). Functional foods. Cancer Letters, 114, 59– 63.
[4] Kitts, D.D., Wijewickreme, A.N. ve Hu, C. (2000). Antioxidant properties of a North American ginseng extract. Molecular Cell Biochemistry, 203, 1- 10.
[5] Lee, J. C. ve Lim, K. T. (2001). Inhibitory effects of the ethanol extract of Ulmus davidiana on apoptosis induced by glucose– glucose oxidase and cytokine production in cultured mouse primary immune cells. Journal of Biochemistry Molecular Biology, 34, 463– 471.
[6] Pezutto, J. M. (1997). Plant-derived anticancer agents. Biochemecial Pharmacolology, 53, 121–133.
[7] Christou, L., Hatzimichael E., Chaidos, A., Tsiara, S.ve Bourantas, K. L. (2001). Treatment of plasma cell leukemia with vincristine, liposomal doxorubicin and dexamethasone. European Journal of Hematology, 67, 51- 53.
[8] Mukherjee, A. K., Basu, S., Sarkar, N. ve Ghosh, A. C. (2001). Advances in cancer therapy with plant-based natural products. Current Medicinal Chemistry, 8, 1467- 1486.
[9] Smets, L. A. (1994). Programmed cell death (apoptosis) and response to anticancer Drugs. Anticancer Drugs, 5, 3-9.
[10] Paschka, A. G., Butler, R. ve Young, C. Y. F. (1998). Induction of apoptosis in prostate cancer cell lines by the green tea component, Epigallocatechin-3- Gallate. Cancer Letters, 130, 1-7.
[11] Cotelle, N. (2001). Role of flavonoids in oxidative stres. Curr. Top. Med. Chem., 1, 569-590.
[12] Wisura,W. ve Glen, H. F. (1993). The South African species of Carpobrotus (Mesembryanthema-Aizoaceae). Contribution Bolus Herbal, 15, 76–107.
[13] Van der Watt, E. ve Pretorius, J. C. (2001). Short communication: Purification and identification of active antibacterial components in Carpobrotus edulis. Journal of Ethanopharmacology, 76, 87-91.
[14] Springfield, E. P., Amabeoku, G., Weitz, F., Mabusela, W. Ve Jhonson, Q. (2003). An assessement of two Carpobrotus species extracts as potential antimicrobial agents. Phytomedicine, 10, 434-439.
[15] Ravishankara M. N. Neeta, S., Harish, P. ve Rajani, M. (2002). Evaluation of antioxidant properties of root bark of Hemidesmus indicus R. Br. (Anantmul). Phytomedicine, 9, 153-160.
[16] Dominguez, X. A. 1973. Metodos de investıgacion fitoquimica. In: Bolivar, P., Cruz-Peredes, C., Kernandez, L. R., Juarez, Z. N., Sanchez-Arreola, E., Av-Gay, Y. ve Bach, H. (2011). Antimicrobial, anti-inflammatory, and cytotoxic activities of Galium mexicanum. Journal of Ethnopharmacology, 137, 141-147.
[17] Singleton,V. L., Orthofer, R.ve Lamuela-Raventõs, 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. [18] Brand-Williams,W., Cuvelier,M. E. ve Berset, C. (1995). Use of a free radical method to evaluate antioxidant activity. Food Science and Technology, 28, 25-30.
[19] Ruch R. J., Cheng, S. J. ve Klaunig, J. E. (1989). Prevention of cyto-toxicity and inhibition of intercellular communication by antioxidant catechins isolated from Chinese green tea. Carcinogenesis, 10(6), 1003-1008.
[20] Dinis, T. C. P., Madeira, V. M. C. ve Almedia, L. M. (1994). Action of phenolic derivatives (Acetaminophen, salicylate, and 5-aminosalicylate) as inhibitors of membrane lipid peroxidation and as peroxyl radical scavengers. Archives of Biochemistry and Biophysics, 315(1), 161-169.
[21] Mossman, T. (1983). Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J. Immunol. Methods, 65, 55-63.
[22] Singh, N. P. (2005). Apoptozis assessment by the DNA diffusion assay. Methods in Molecular Medicine, 111, 55-67. [23] Arıduru, R. ve Arabacı, G. (2013). Ciğertaze otu (Salvia officinalis) bitkisinin antioksidan aktivitesinin belirlenmesi. SAÜ. Fen Bilimleri Dergisi, 17( 2), 241-246.
[24] Wojdylo, A., Oszmianski, J. ve Czmerys, R. (2007). Antioxidant activity and phenolic compounds in 32 selected herbs. Food Chem., 105, 940-949.
[25] Chen, T., Mei, N. ve Fu, P. P. (2010). Genotoxicity of pyrrolizidine alkaloids. Journal of Applied Toxicology, 30, 183-196.
[26] Falleh, H., Ksouri, R., Medini, F., Guyot, S., Abdelly, C., Christian, M. (2011). Antioxidant activity phenolic composition of the medicinal and edible halophyte Mesembryanthemum edule. L. Industrial Crops and Products. 34. 1066-1071
[27] Miliauskas, G., Venskutonis, P. R. ve Van Beek, T. A. (2004). Screening of radical scavenging activity of some medicinal and aromatic plant extracts. Food Chemistry, 85, 231-237.
[28] Vundać, V. B., Brantner, A. H. ve Plazibat, M. (2007). Content of polyphenolic constituents and antioxidant activity of some Stachys taxa. Food Chemistry, 104, 1277-1281.
[29] Aslantürk, Ö. S. (2010). Aydın Yöresinde Kullanılan Bazı Tıbbi Bitkilerin Antioksidant ve Sitotoksik Etkilerinin Araştırılması. Doktora Tezi, Adnan Menderes Üniversitesi Fen Bilimleri Enstitüsü, Aydın.
[30] Aslantürk, Ö.S. ve Aşkın Çelik, T. A. (2013). Antioxidant activity and anticancer effect of Vitex agnus-castus L. (Verbenaceae) seed extracts on MCF-7 breast cancer cells. Caryologia, 66(3), 257-267.
[31] Doğmuş, D. ve Durucasu, İ. (2013). Keten tohumu çeşitlerinin N-Bütanol fraksiyonlarının fenolik bileşenlerinin antioksidan aktivitesi. C.B.Ü. Fen Bilimleri Dergisi, 9(1), 47 - 56.
[32] Halliwell, B. ve Gutteridge J.M.C. (1990). Role of free radicals and catalytic metal ions in human disease: An overview. Methods in Enzymology, 186, 1-85.
[33] Arunporn, I., Peter, J., Houghton, E. ve Amooquaye, E. (2004). In vitro cytotoxic activity of Thai medicinal plants used traditionally to treat cancer. J. Ethnopharmacol., 90, 33-38.
[34] Bartek, J., Lukas, C. ve Lukas, J. (2004). Checking on DNA damage in S phase. Nature Reviews Molecular Cell Biology, 5, 792-804.
[35] Thompson, C. B. (1995). Apoptosis in the pathogenesis and treatment of disease. Science, 267, 1456-1462. [36] Kinloch, R. A., Trekerne, J. M., Furness, L. M. ve Hajimohamadreza, I. (1999). The pharmacology of apoptosis. Trends in Pharmacology Science, 20, 35-42.
[37] Martins, A., Vasas, A., Schelz, M., Viveiros, M., Molnár; J., Hohman, J., Amaral, L. (2010). Constituents of Carpobrotus edulis inhibit P-glycoprotein of MDR-1transfected Mouse lymphoma cells. Anticancer Research. 30.829-836.
[38] Kwon, K. H., Barve, A., Yu, S., Huang, M.T. ve Kong, A.N.T. (2007). Cancer chemoprevention by phytochemicals: potential molecular targets, biomarkers, and animal models. Acta Pharmacol. Sin., 28, 1409-1421.
[39] Han, X. Z., Shen, T. ve Lou, H. X. (2007). Dietary polyphenols and their biological significance. Int. J. Mol. Sci., 8, 950-988.
[40] Fresco, P., Borges, F., Diniz, C. ve Marques, M.P. (2006). New insights on the anticancer properties of dietary polyphenols. Med. Res. Rev., 26, 747-766.
[41] Garuti, L., Roberti, M. ve Pizzirani, D. (2005). Nitrogen-containing heterocyclic quinones: a class of potential selective antitumor agents. Mini-Review Medicinal Chemistry, 7, 481-489.
[42] Demirezer, L. O., Kuruüzüm-Uz, A., Bergere, I., Schiewe, H. J. ve Zeeck, A. (2001). The structures of antioxidant and cytotoxic agents from natural saurce: anthraquinones and tannins from roots of Rumex patientia. Phytochemistry, 58, 1213-1217.
[43] Chang, Y. C., Tai, K. W., Huang, F. M., Huang, M. F. 2003. Cytotoxic and nongenotoxic effects of phenolic compounds in human pulp cell cultures. J. Endodontics, 26(8): 440-443.