Alıç ve Mürver Ekstraktlarının 3T3 Fibroblast Hücre Hattı Üzerindeki Sitotoksik Etkileri

Öz

Bitkiler, eski zamanlardan beri sayısız insan tarafından tıbbi ilaçlarda kullanılmaktadır. Şifalı bitkilerin çok çeşitli hastalıklar için birçok faydası vardır. Günümüzde, bilim ve tıbbi bitkiler birlikte ilerleyebilmektedir. Bu çalışmada, alıç (Crataegus oxyacantha) ve mürver (Sambucus nigra) bitkileri kullanılmaktadır. Kalp ve damar hastalıklarının tedavisinde, göğüs ağrısı, düzensiz kalp atışı, sindirim sorunları ve kolesterolün düzenlenmesinde alıç bitkisinden yararlanıldığı bildirilmektedir. Bunların yanısıra, iltihaplanma, ateş, eklem ve kas ağrısı, basit cilt problemleri, stres ve hatta HIV tedavisinde mürver bitkisinin kullanıldığı iddia edilmektedir. Bu çalışmada, 3T3 fare kökenli fibroblast hücre hattı kullanılarak farklı konsantrasyonlarda alıç ve mürver ekstraktlarının sitotoksisite araştırmaları gerçekleştirilmiştir. 1 x 104 hücre / kuyu miktarda hücre, 96 kuyucuklu plaka içine eşit olarak ekilerek yarı konfluent tek katman hücre oluşumu için 24 saat inkübe edilmiştir. Kuyucuk başına çeşitli konsantrasyonlarda alıç ve mürver ekstraktları (10, 5, 1, 0.5, 0.1, 0.05 ve 0.01 mg / ml) içeren ortam ilave edilerek inkübasyon süreci devam ettirilmiştir. Canlılık analizi, WST-1 kolorimetrik analiz kiti kullanılarak 48 saat sonra gerçekleştirilmiştir. Sonuç olarak, analizi gerçekleştirilen konsantrasyon oranlarının toksik özellik göstermediği ve iki ekstrakt için de 0.05 mg / ml konsantrasyon kullanılmasıyla daha iyi çoğalma olabileceği kanısına varılmıştır.

Anahtar Kelimeler

• Bitki ekstraktları,Alıç,Mürver,Sitotoksisite,WST-1.

Cytotoxic Effects of Hawthorn and Elderberry Extracts on 3T3 Fibroblast Cell Line

Abstract

Plants have been used in the medical remedies by countless people since the ancient times. Medicinal plants have multiple benefits for various cases. Nowadays, science is developing in correlation with these medicinal plants. Hawthorn (Crataegus oxyacantha) and elderberry (Sambucus nigra) is used in this study as a medicinal plant. Hawthorn has been claimed to be used in the treatment of heart and blood vessel diseases, chest pain, irregular heartbeat, digestive problems and cholesterol regulation. On the other hand, elderberry has been claimed to be used in inflammation, fever, joint and muscle pain, minor skin conditions, stress and even HIV. In this study, we investigated the cytotoxicity levels of the hawthorn and elderberry extracts with different concentrations on 3T3 murine fibroblast cell line. 1 × 104 cells/well were equally seeded in 96 well plate and incubated for 24 hours to form a half-confluent monolayer. Treatment medium containing various concentrations of extracts (10, 5, 1, 0.5, 0.1, 0.05 and 0.01 mg/ml) were added per well. The viability analysis was conducted after 48 hours by using the WST-1 colorimetric assay. In conclusion, each concentration used is non-cytotoxic and better proliferation activities were reached by using 0.05 mg/ml concentration for hawthorn and elderberry. 

Keywords

• Plant extracts,Hawthorn,Elderberry,Cytotoxicity,WST-1.

Kaynakça

1. 1. Historical review of medicinal plants' usage. Petrovska, BB. 11, Jan 2012, Pharmacogn Rev., Cilt 6, s. 1-5.
2. 2. A SHORT REVIEW ON THE MEDICINAL PLANTS AND THEIR USES. Jan, Ghulam Mohammad. 5, 2012, World Journal of Pharmaceutical Research, Cilt 1, s. 1518-1521.
3. 3. Hawthorn: a literature review. Hobbs C, Foster S. 19, 1994, Herbal Gram, Cilt 22, s. 19-33.
4. 4. Heinerman J: West Nyack, NY. Heinerman’s Encyclopedia ofHealing Herbs and spices. [kitap yaz.] Heinerman J. 1996, s. 19-33.
5. 5. Hawthorn. Qi Chang, Zhong Zuo, Francisco Harrison, and Moses Sing Sum Chow. 2002, Journal of Clinical Pharmacology, Cilt 42, s. 605-612.
6. 6. Hawthorn: potential roles in cardiovascular disease. W.-T. Chang, J. Dao, and Z.-H. Shao. 1, 2005, The American Journal of Chinese Medicine, Cilt 33, s. 1-10.
7. 7. Botanical influences on cardiovascular disease,. Miller, A. L. 6, 1998, Alternative Medicine Review, Cilt 3, s. 422-431.
8. 8. Effect of Crataegus Usage in Cardiovascular Disease Prevention: An Evidence-Based Approach. Jie Wang, Xingjiang Xiong, and Bo Feng. 2013, Herbal Medicines for Cardiovascular Diseases.

9. 9. European elderberry (Sambucus nigra L.) rich in sugars, organic acids, anthocyanins and selected polyphenols. Robert Veberic, Jerneja Jakopic, Franci Stampar, Valentina Schmitzer. 2009, Food Chemistry , Cilt 114, s. 511-515.
10. 10. Analytical characterization and the impact of ageing in anthocyanin composition and degradation in juices from five sour cherry cultivars. . Bonerz, D., Würth, K., & Dietrich, H. 2006, European Food Research and Technology, Cilt 224, s. 355–364.
11. 11. Steam-blanched highbusch blueberry (Vaccinium corymbosum L.) juice: Phenolic profile and antioxidant capacity in relation to cultivar selection. Brambilla, A., Lo Scalzo, R., Bertolo, G., & Torreggiani, D. 2008, Journal of Agricultural and Food Chemistry, Cilt 56, s. 2643–2648.
12. 12. Apple peel as a contributor to whole fruit quantity of potential healthful bioactive compounds. Cultivar and year implication. Lata, B., & Tomala, K. 2007, Journal of Agricultural and Food Chemistry, Cilt 55, s. 10795–10802.
13. 13. Measuring Cell Death by Trypan Blue Uptake and Light Microscopy. Crowley L.C., Marfell B.J., Christensen M.E. ve Waterhouse N.J. 2016, Cold Spring Harb Protoc., Cilt 7.
14. 14. WST-1-based cell cytotoxicity assay as a substitute for MTT-based assay for rapid detection of toxigenic Bacillus species using CHO cell line. Ngamwongsatit P, Banada PP, Panbangred W, Bhunia AK. 2008, Journal of Microbiological Methods, Cilt 73, s. 211-215.
15. 15. Biological effects of imidazolium ionic liquids with varying chain lengths in acute Vibrio fischeri and WST-1 cell viability assays. Ranke J, Molter K, Stock F, Bottin-Weber U, Poczobutt J, Hoffmann J, Ondruschka B, Filser J, Jastorff B. Ecotoxicol Environ Safety : yazarı bilinmiyor, 2004, Cilt 58, s. 396–404.
16. 16. Autophagy modulates endoplasmic reticulum stress-induced cell death in podocytes: A protective role. . Cheng, Y.-C., Chang, J.-M., Chen, C.-A., & Chen, H.-C. 4, 2014, Experimental Biology and Medicine, Cilt 240, s. 467–476.
17. 17. Effects of extracts of Salvadora persica on proliferation and viability of human dental pulp stem cells. Fahimeh sadat Tabatabaei, Maryam Moezizadeh, Fateme Javand. 4, 2015, J Conserv Dent., Cilt 18, s. 315-320.
18. 18. Ali-Boucetta H., Al-Jamal K.T., Kostarelos K. Cytotoxic Assessment of Carbon Nanotube Interaction with Cell Cultures. [kitap yaz.] Hurst S. Methods in Molecular Biology. basım yeri bilinmiyor : Humana Press, 2011, s. 299-312.
19. 19. ISO. Biological Evaluation of Medical Devices. Tests for in vitro Cytotoxicity. 1999. ISO B. 10993-5.
20. 20. BioVision. Ready-to-use Cell Proliferation Colorimetric Reagent, WST-1. [Çevrimiçi] https://www.biovision.com/documentation/datasheets/K304.pdf.
21. 21. Aslantürk, Özlem Sultan. In Vitro Cytotoxicity and Cell Viability Assays: Principles, Advantages, and Disadvantages. Genotoxicity: A Predictable Risk to Our Actual World. 2017.
22. 22. Differential effects of polyphenols-enriched extracts from hawthorn fruit peels and fleshes on cell cycle and apoptosis in human MCF-7 breast carcinoma cells. Ting Li, Jiao Zhu, Ling Guo, Xiaolong Shi, Yafei Liu, Xingbin Yang. 2, 2013, Food Chemistry, Cilt 141, s. 1008-1018.
23. 23. Major triterpenoids in Chinese hawthorn “Crataegus pinnatifida” and their effects on cell proliferation and apoptosis induction in MDA-MB-231 cancer cells. Lingrong Wen, Ruixue Guo, Lijun You, Arshad Mehmood Abbasi, Tong Li, Xiong Fu, Rui Hai Liu. 2017, Food and Chemical Toxicology, Cilt 100, s. 149-160.
24. 24. Triterpenoid Acids as Important Antiproliferative Constituents of European Elderberry Fruits. Michał Gleńsk, Elżbieta Czapińska, Marta Woźniak, Ireneusz Ceremuga, Maciej Włodarczyk, Grzegorz Terlecki, Piotr Ziółkowski & Ewa Seweryn. 4, 2017, Nutrition and Cancer, Cilt 69, s. 643-651.
25. 25. Hypotensive effects of hawthorn for patients with diabetes taking prescription drugs: a randomised controlled trial. Ann F Walker, Georgios Marakis, Eleanor Simpson, Jessica L Hope, Paul A Robinson, Mohamed Hassanein and Hugh CR Simpson. 56, 2006, British Journal of General Practice., Cilt 527, s. 437-443.
26. 26. Advanced research on the antioxidant and health benefit of elderberry (Sambucus nigra) in food – a review. Andrzej Sidor, Anna Gramza-Michałowska. Part B, 2015, Journal of Functional Foods, Cilt 18, s. 941-958.
27. 27. Protective intervention of Sambucus nigra polyphenols in the diabetic heart. . Ciocoiu, M. & Badescu, L. & Badulescu, O. & Tutunaru, Dana & Badescu, M. 2012, Annals of the Romanian Society for Cell Biology., Cilt 17, s. 312-317.
28. 28. Intervention of Sambucus Nigra Polyphenolic Extract in Experimental Arterial Hypertension. Manuela Ciocoiu, Laur Badescu, Oana Badulescu, Magda Badescu. 4, 2012, International Journal of Medical and Health Sciences, Cilt 6.
29. 29. Hawthorn (Crataegus spp.) in the treatment of cardiovascular disease. Tassell, Mary & Kingston, Rosari & Gilroy, Deirdre & Lehane, Mary & Furey, Ambrose. 2010, Pharmacognosy reviews. , Cilt 4, s. 32-41.