Rize ve Çevresinde Yetişen Yaban Meyvelerinden Elde Edilen Özütlerin Anti-Proliferatif Etkilerinin Araştırılması

Year 2019, Volume: 3 Issue: 3, 104 - 111, 03.12.2019

Saliha Ekşi Sena Şahin Aktura Nebahat Ejder Kazım Şahin

Abstract

ÖZ

Amaç: Doğu Karadeniz bölgesi, farklı tıbbi ve aromatik bitkileri barındıran
zengin bir floraya sahiptir. Rize ve çevresinde başta Arbutus unedo
(Kocayemiş), Aronia melanocarpa (Rus yaban mersini), Fragaria vesca
L. (Yabani çilek), Frangula alnus (Barut ağacı), Ribes rubrum
(Frenk üzümü) ve Solanum nigrum (Köpek üzümü) olmak üzere çeşitli
meyveler doğal olarak yetişmektedir. Bu çalışmada, adı geçen meyvelerden elde
edilen metanol özütlerinin insan retina pigment epitel hücreleri (ARPE-19) ve
insan serviks adenokarsinoma hücreleri (HeLa) üzerindeki antiproliferatif
etkinliklerinin değerlendirilmesi amaçlandı.

Araçlar
ve Yöntem: Meyvelerin metanol özütleri elde edilerek, çeşitli
konsantrasyonlarda ARPE-19 ve HeLa hücre kültür ortamlarına eklendi. Pozitif ve
negatif kontrol olarak sırasıyla, Taksol ve DMSO kullanıldı. 48 saatlik
inkübasyon sonrası özütlerin hücreler üzerindeki sitotoksik etkileri MTT testi
ile değerlendirildi. İstatistiksel analizler bağımsız örneklem t-testi
kullanılarak gerçekleştirildi. p değeri 0.05’ten küçük olan sonuçlar anlamlı
olarak kabul edildi.

Bulgular: R.rubrum ve S.nigrum özütleri iki hücre hattında da doza
bağımlı sitotoksik aktivite göstermişler, buna ek olarak A.melanocarpa
ve F.vesca özütleri yüksek konsantrasyonlarda her iki hücre hattı
üzerinde sitotoksik etki göstermişlerdir. A.unedo ve F.alnus
metanol özütlerinin sadece HeLa üzerinde doza bağımlı olarak hücre canlılığını
inhibe ettiği tespit edilmiştir.

Sonuç: A.unedo, A.melanocarpa, F.vesca, F.alnus, R.rubrum
ve S.nigrum’un hem ARPE-19 hem de HeLa üzerindeki sitotoksik etkileri
ilk defa bu çalışmada araştırılmıştır. A.unedo ve F.alnus  bitki ekstrelerinin  antiproliferatif ve sekonder bileşenlerini
belirlemek için daha fazla çalışmaya ihtiyaç duyulmaktadır. 

Keywords

Serviks kanseri, sitotoksisite, Arbutus unedo, Frangula alnus, metiltiazol difenil tetrazolyum

Supporting Institution

Recep Tayyip Erdoğan Üniversitesi

Project Number

BAP-2015.53001.106.01.04

Thanks

Bu çalışma Recep Tayyip Erdoğan Üniversitesi BAP-2015.53001.106.01.04 nolu proje ile desteklenmiştir. Çalışmada kullanılan HeLa hücre hattını bize temin eden Prof. Dr. Muradiye Acar’a ve ARPE-19 hücre hattını sağlayan Prof. Dr. Fikrettin Şahin’e teşekkür ederiz.

Investigation of Anti-Proliferative Activities of Extracts Derived From Wild Fruits That Grow in Rize Vicinity

Abstract

ABSTRACT

Purpose: Eastern Black Sea region has a rich flora with
different medicinal and aromatic plants. In particular, Arbutus unedo (Strawberry tree), Aronia melanocarpa (Russian bilberry), Fragaria vesca L. (Wild strawberry), Frangula alnus (Alder dogwood), Ribes
rubrum (Red currant) and Solanum
nigrum (Nightshade), various fruits grow indigenous to Rize and its
vicinity. The aim of the study was to evaluate the anti-proliferative activity
of these fruits on human retinal pigment epithelial cells (ARPE-19) and human
cervix adenocarcinoma cells (HeLa). 

Materials and Methods: Methanol extracts of the
fruits were obtained and added to ARPE-19 and HeLa culture medias at various
concentrations. Taxol and DMSO were used as positive and negative controles,
respectively. After 48 hours of cell incubation, the cytotoxic effect of the
extracts was assessed by MTT. Cell viability values ​​were calculated for each
group. The statistical analysis of the data was performed using unpaired
t-test. p values ​​less than 0.05 were considered significant.

Results: The extracts of R.rubrum and S.nigrum  showed dose dependent cytotoxic activity
against both cell lines. In additions, A.melanocarpa
and F.vesca  extracts exhibited cytotoxic effect on
both cell lines at higher concentrations. Methanol extracts obtained from A.unedo and F.alnus  were found to decrease cell viability in a
dose-dependent manner only in HeLa cells.

Conclusion: The cytotoxic effects of A.unedo, A.melanocarpa, F.vesca,
F.alnus, R.rubrum  ande S.nigrum’un  on both ARPE-19 and HeLa
were firstly investigated in this study.
To underline the antiproliferative effects of. A.unedo  and F.alnus   extracts as well as their seconder  components further studies are needed. 

Keywords

Cervical cancer, cytotoxicity, Arbutus unedo, Frangula alnus, methylthiazole diphenyl tetrazolium

Project Number

BAP-2015.53001.106.01.04

References

• Ceyhan M. İnsan papilloma virusu (HPV) aşısı uygulamasında ülkemizde mevcut problemler. ANKEM Dergisi, 2007; 21(2): 102-104.
• Uysal A, Zengin G, Durak Y, Aktümsek A. Centaurea pterocaula özütlerinin antioksidan ve antimutajenik özellikleri ile enzim inhibitör potansiyellerinin incelenmesi. Marmara Pharmaceutical Journal, 2016;20: 232-242.
• Zengin G, Uysal A, Gunes E, Aktumsek A. Survey of Phytochemical Composition and Biological effects of Tree extracts from a Wild Plant (Cotoneaster nummularia Fisch et mey.): A Potential Source for Functional Food Ingredients and Drug Formulations. Plos One 2014; 9: 1-13.
• Akin D, Durak Y, Uysal A, Gunes E, Aladag MO. Assessment of Antimutagenic Action of Celtis glabrata Steven ex Planch. (Cannabaceae) extracts Against Base Pair exchange and Frame Shif mutations on Salmonella typhimurium tA98 and tA100 Strains by Ames test. Drug Chem toxicol 2016; 39:312-21.
• Uysal A, Gunes E, Sarikurkcu C, Celik H, Durak Y, Uren MC. New Prospective Materials for Chemoprevention: Tree Phlomis. Brit J Pharm res 2016; 10:1-13.
• Miguel MG, Faleiro ML, Guerreiro AC, Antunes MD. Arbutus unedo L.: chemical and biological properties. Molecules, 2014; 19(10), 15799-15823.
• Barros L, Carvalho AM, Morais JS, Ferreira ICFR. Strawberry-tree, blackthorn and rose fruits: Detailed characterisation in nutrients and phytochemicals with antioxidant properties. Food Chemistry, 2010; 120, 247–254.
• Özcan MM, Hacıseferioğulları H. The Strawberry (Arbutus unedo L.) fruits: Chemical composition,physical properties and mineral contents. Journal of Food Engineering, 2007; 78, 1022–1028.
• Guimaraes R, Barros L, Duenas M, Carvalho AM, Queiroz MJRP, Santos-Buelga C et al. Characterisation of phenolic compounds in wild fruits from Northeastern Portugal. Food Chemistry, 2013; 141, 3721–3730.
• Wawer I, Wolniak M, Paradowska K. Solid state NMR study of dietary fiber powders from aronia, bilberry, black currant and apple. Solid state nuclear magnetic resonance, 2006; 30(2), 106-113.
• Ciocoiu M, Badescu L, Miron A, Badescu M. The involvement of a polyphenol-rich extract of black chokeberry in oxidative stress on experimental arterial hypertension. Evidence-Based Complementary and Alternative Medicine, 2013.
• Turhan E, Kargı SP. Strawberry Production in Turkey. Chronica Horticulturae, 2007; 47(2):18–20.
• Doğanoğlu Ö, Gezer A, Yücedağ C. Göller Bölgesi-Yenişarbademli Yöresi’nin Önemli Bazı Tıbbi ve Aromatik Bitki Taksonları Üzerine Araştırmalar, Süleyman Demirel Üniversitesi Fen Bilimleri Enstitüsü Dergisi, 2006; 10-1,66-73.
• McDougall GJ, Ross HA, Ikeji M, Stewart D. Berry Extracts Exert Different Antiproliferative Effects against Cervical and Colon Cancer Cells Grown in Vitro, Journal of Agriculture and Food Chemistry, 2008; 5693016-3023.
• Dara R, Vefalı Dostlarım Şifalı Otlarım. Alfa Yayınları, No:1665, İstanbul, Türkiye, 2006.
• Kremer D, Kosalec I, Locatelli M, Epifano F, Genovese S, Carlucci G et al. Anthraquinone profiles, antioxidant and antimicrobial properties of Frangula rupestris (Scop.) Schur and Frangula alnus Mill. Bark. Food Chem., 2012; 131:1714-1180.
• Heywood VH, Brummıtt RK, Culham A, Seberg O. Flowering Plant Families of The World. Firefly Books pres, Ontario, Canada, 2007; 160.
• Singh A, Lal M, Samant SS. Diversity, indigenous uses and conservation prioritization of medicinal plants in Lahaul valley, proposed cold desert biosphere reserve, India. International Journal of Biodiversity Science, Ecosystem Services & Management, 2009; 5(3): 132-154.
• Šarıć-Kundalıć B, Dobeš C, Klatte-Asselmeyer V, Saukel J. Ethnobotanical study on medicinal use of wild and cultivated plants in middle, South and west Bosnia and Herzegovina. Journal of Ethnopharmacology, 2010; 131(1): 33-55.
• Ronald A. The etiology of urinary tract infection: traditional and emerging pathogens. Am. J. Med., 2002; 113(1A): 14-19.
• Güvenç A, Houghton PJ, Duman H, Coşkun M, Şahin P. Antioxidant activity studies on selected Sideritis species native to Turkey. Pharmaceutical Biology, 2005; 43(2): 173-177.
• Tanker N, Koyuncu M, Coşkun M. Solanaceae, Farmasötik Botanik, Ankara Üniversitesi Eczacılık Fakültesi Yayınları, Ankara, 2007; 296-301.
• Dhellot JR, Matouba E, Maloumbi MG. Extraction and Nutritional Properties of Solanum nigrum L. Seed Oil, African Journal of Biotechnology, 2006; 5(10), 987-991.
• Mossman T. Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. Journal of lmmunological Methods, 1983; 65: 55-63.
• Yin X, Zhou J, Jie C, Xing D, Zhang Y. Anticancer activity and mechanism of Scutellaria barbata extract on human lung cancer cell line A549 Life Sciences., 2004;75-. (18):2233-2244.
• Benzie IFF, Wachtel-Galor S. Herbal Medicine: Biomolecular and Clinical Aspects. 2. Baskı, CRC Press, 2011, Boca Raton ,Florida.
• Christou L, Hatzimichael E, Chaidos A, Tsiara S, Bourantas KL. Treatment of plasma cell leukemia with vincristine, liposomal doxorubicin and dexamethasone. European Journal of Hematology, 2001; 67: 51-53.
• Mukherjee AK, Basu S, Sarkar N, Ghosh AC. Advances in cancer therapy with plant-based natural product. Current Medicinal Chemistry, 2001; 8: 1467-1486.
• Afrin S, Forbes-Hernandez TY, Gasparrini M, Bompadre S, Quiles JL, Sanna G et al. Strawberry-Tree Honey Induces Growth Inhibition of Human Colon Cancer Cells and Increases ROS Generation: A Comparison with Manuka Honey. Department of Clinical Sciences, Faculty of Medicine, Polytechnic University of Marche, via Ranieri 65,Ancona 60131, Italy, 2016.
• Siegers CP, von Hertzberg-Lottin E, Otte M, Schneider B. Anthranoid laxative abuse a risk for colorectal cancer, 1993.
• Kupchan SM, Karim A. Tumor inhibitors. 114. Aloe emodin: antileukemic principle isolated from Rhamnus frangula L., 1976.
• Thani NA, Keshavarz S, Lwaleed BA, Cooper AJ, Rooprai HK. Cytotoxicity of gemcitabine enhanced by polyphenolics from Aronia melanocarpa in pancreatic cancer cell line AsPC-1. J Clin Pathol. 2014; Nov;67(11):949-54.
• Bin L, Enhui L, Shurui C, Xinyao J, Yuehua W, Ningxuan G. Preparative Purification of Polyphenols from Aronia melanocarpa (Chokeberry) with Cellular Antioxidant and Antiproliferative Activity, 2017.