BUĞDAY ÇİMİNİN İNSAN LENFOSİT HÜCRELERİ ÜZERİNE ETKİSİ

Year 2023, Volume: 30 Issue: 1, 47 - 55, 14.03.2023

Okan Sancer , Zehra Safi Öz , Pınar Aslan Koşar

https://doi.org/10.17343/sdutfd.1240777

Abstract

Amaç
Kemoterapötik ilaçlar kanser hücrelerinin ortadan
kaldırılmasında etkili iken aynı zamanda sağlıklı hücrelerde
de hasar oluşturabilmektedir. Bu çalışmada,
buğday çimi (Triticum aestivum L.) ekstraktının fenolik
bileşen içeriğinin analizi ve bu ekstraktın kemoterapötik
tedavide kullanılan sisplatin ve etoposid’in sağlıklı
hücrelerde oluşturduğu DNA hasarına karşı etkisinin
belirlenmesi amaçlanmıştır.
Gereç ve Yöntem
Çimlendirilmiş buğdayların metanol ekstraktı hazırlanarak
HPLC (yüksek performanslı sıvı kromatografisi)
ile fenolik bileşen analizi yapıldı. Buğday ekstraktı
konsantrasyonuna bağlı hücre canlılık testi uygulanarak
IC50 (Yarı maksimum inhibitör konsantrasyonu) ve
LD50 (ortalama öldürücü doz) değerleri hesaplandı.
Belirlenen bu konsantrasyon değerleri ile hücreler inkübe
edilerek DNA hasarı varlığı Comet metodu ile
değerlendirildi.
Bulgular
Fenolik bileşen analizi sonucunda p-hidroksibenzoik
asit en yüksek miktarda, o-kumarik asit ise en düşük
düzeyde tespit edildi. Lenfosit hücrelerine uygulanan
farklı konsantrasyonlardaki buğday çimi ekstraktı,
etoposid ve sisplatin için değerler sırasıyla IC50=204,6
μg/mL, LD50=15,84 μg/mL ve LD50=24,51 μg/mL olarak
bulundu. Comet analizi sonucunda kontrol grubuna
kıyasla, etoposid LD50 ve etoposid LD50+buğday
çimi ekstraktı IC50 grubu istatistiksel olarak anlamlı
bulunurken (p<0,05), etoposid LD50 ve etoposid
LD50+buğday çimi ekstraktı IC50 grubu arasında istatistiksel
olarak anlamlılık bulunamadı (p>0,05). Bu
sonuca benzer olarak kontrol grubuna kıyasla, sisplatin
LD50 ve sisplatin LD50+buğday çimi ekstraktı IC50
grubu istatistiksel olarak anlamlı bulunurken (p<0,05),
etoposid LD50 ve etoposid LD50+buğday çimi ekstraktı
IC50 grubu arasında istatistiksel olarak anlamlılık saptanmadı
(p>0,05).
Sonuç
Çalışmamızda buğday çiminin etoposid ve sisplatin
nedeni ile oluşan DNA hasarında azalmaya neden olduğu
görülmüş olmasına rağmen istatistiksel olarak
anlamlılık saptanmamıştır.

Keywords

Buğday çimi, Comet metodu, HPLC, MTT

Thanks

Çalışmamıza ait bazı deneysel aşamaların gerçekleştirilmesinde cihaz kullanılması ile destek olan Süleyman Demirel Üniversitesi Yenilikçi Teknolojiler Uygulama ve Araştırma Merkezi (YETEM)'e teşekkür ederiz.

EFFECT OF WHEATGRASS ON HUMAN LYMPHOCYTE CELLS

Abstract

Objective
While chemotherapeutic drugs are effective at eliminating
cancer cells, they can also damage healthy
cells. The aim of this study was to analyze the phenolic
component content of wheatgrass extract (Triticum
aestivum L.) and to determine the effect of this
extract against DNA damage caused by cisplatin and
etoposide used in chemotherapeutic treatment in
healthy cells will.
Material and Method
Sprouted wheat methanol extract was prepared and
analysis of phenolic components was performed by
HPLC (high performance liquid chromatography).
IC50 (half maximal inhibitory concentration) and LD50
(median lethal dose) values were calculated by applying
a cell viability assay based on wheat extract
concentration. Cells were incubated at these determined
concentration levels and the presence of DNA
damage was assessed by the Comet method.
Results
As a result of the phenol component analysis, p-hydroxybenzoic
acid was determined in the highest
amount and o-coumaric acid in the lowest amount.
The values for wheatgrass extract, etoposide and
cisplatin at different concentrations applied to lymphocyte
cells were found to be IC50=204.6 μg/mL,
LD50=15.84 μg/mL and LD50=24.51 μg/mL, respectively.
As a result of comet analysis, it was found
that etoposide LD50 and etoposide LD50+wheatgrass
extract IC50 group were statistically significant
(p<0.05), compared to the control group, there was
no statistical significance between etoposide LD50
and etoposide LD50+wheatgrass extract IC50 group
(p>0.05). Similar to this result, cisplatin LD50 and
cisplatin LD50+wheatgrass extract IC50 group were
found to be statistically significant compared to the
control group (p<0.05), while there was no statistical
significance between etoposide LD50 and etoposide
LD50+wheatgrass extract IC50 group (p> 0.05).
Conclusion
Although wheatgrass was observed in our study to
cause a reduction in DNA damage caused by etoposide
and cisplatin, no statistical significance was
found.

Keywords

Wheatgrass, Comet method, HPLC, MTT

References

• 1. Rommasi F, Esfandiari N. Liposomal nanomedicine: applications for drug delivery in cancer therapy. Nanoscale Research Letters. 2021;16(1):1-20.
• 2. Sun C-Y, Zhang Q-Y, Zheng G-J, Feng B. Phytochemicals: Current strategy to sensitize cancer cells to cisplatin. Biomedicine & Pharmacotherapy. 2019;110:518-27.
• 3. Fang C-y, Lou D-y, Zhou L-q, Wang J-c, Yang B, He Q-j, et al. Natural products: Potential treatments for cisplatin-induced nephrotoxicity. Acta Pharmacologica Sinica. 2021;42(12):1951- 69.
• 4. Tchounwou PB, Dasari S, Noubissi FK, Ray P, Kumar S. Advances in our understanding of the molecular mechanisms of action of cisplatin in cancer therapy. Journal of experimental pharmacology. 2021;13:303.
• 5. Kim SS, Wengier DL, Ragland CJ, Sattely ES. Transcriptional Reactivation of Lignin Biosynthesis for the Heterologous Production of Etoposide Aglycone in Nicotiana benthamiana. ACS synthetic biology. 2022;11(10):3379-87.
• 6. Zhang W, Gou P, Dupret J-M, Chomienne C, Rodrigues-Lima F. Etoposide, an anticancer drug involved in therapy-related secondary leukemia: Enzymes at play. Translational oncology. 2021;14(10):101169.
• 7. Gore RD, Palaskar SJ, Bartake AR. Wheatgrass: Green blood can help to fight cancer. Journal of Clinical and Diagnostic Research: JCDR. 2017;11(6):ZC40.
• 8. Adams M, Jewell A. The use of complementary and alternative medicine by cancer patients. International Seminars in Surgical Oncology. 2007;4:10.
• 9. Hassan N, Siddique MS. Wheat Grass (Triticum aestivum L.) Benefits Health in a Pandemic Scenario. Journal for Research in Applied Sciences and Biotechnology. 2022;1(1):24-9.
• 10. Patel JB. Anticancer & cytotoxic potential of aqueous extract of Triticum aestivum on hela cell line. Journal of Drug Delivery and Therapeutics. 2016;6(3):84-9.
• 11. Kaur N, Singh B, Kaur A, Yadav MP, Singh N, Ahlawat AK, et al. Effect of growing conditions on proximate, mineral, amino acid, phenolic composition and antioxidant properties of wheatgrass from different wheat (Triticum aestivum L.) varieties. Food Chemistry. 2021;341:128201.
• 12. Garg M, Sharma A, Vats S, Tiwari V, Kumari A, Mishra V, et al. Vitamins in cereals: a critical review of content, health effects, processing losses, bioaccessibility, fortification, and biofortification strategies for their improvement. Frontiers in nutrition. 2021;8:254.
• 13. Caponio F, Alloggio V, Gomes T. Phenolic compounds of virgin olive oil: influence of paste preparation techniques. Food Chemistry. 1999;64(2):203-9.
• 14. Singh NP, McCoy MT, Tice RR, Schneider EL. A simple technique for quantitation of low levels of DNA damage in individual cells. Experimental cell research. 1988;175(1):184-91.
• 15. Molaae N, Mosayebi G, Pishdadian A, Ejtehadifar M, Ganji A. Evaluating the proliferation of human PeripheralBlood mononuclear cells using MTT assay. International Journal of Basic Science in Medicine. 2017;2(1):25-8.
• 16. Alitheen NB, Oon CL, Keong YS, Chuan TK, Li HK, Yong HW. Cytotoxic effects of commercial wheatgrass and fiber towards human acute promyelocytic leukemia cells (HL60). Pakistan Journal of Pharmaceutical Sciences. 2011;24(3): 243-250
• 17. Ghumman A, Singh N, Kaur A. Chemical, nutritional and phenolic composition of wheatgrass and pulse shoots. International journal of food science & technology. 2017;52(10):2191-200.
• 18. Rodríguez FC, Gallagher E, Rai DK, Burgess CM. Nutritional and physiochemical properties of wheatgrass juice and preservation strategies. Food Chemistry Advances. 2022:100136.
• 19. Choi M-H, Lee MY, Yang S-H, Shin H-J, Jeon YJ. Hydrophobic fractions of Triticum aestivum L. extracts contain polyphenols and alleviate inflammation by regulating nuclear factor-kappa B. Biotechnology and Bioprocess Engineering. 2021;26(1):93-106.
• 20. Rosa L, Moreno-Escamilla J, Rodrigo-García J, Alvarez-Parrilla E. Phenolic compounds. Postharvest physiology and biochemistry of fruits and vegetables. 2019:253-71.
• 21. Eissa HA, Mohamed SS, Hussein A. Nutritional value and impact of wheatgrass juice (Green Blood Therapy) on increasing fertility in male albino rats. Bulletin of the National Research Centre. 2020;44(1):1-11.
• 22. Hebbani AV, Bulle S, Kanu VR, Balachandrababu Malini A, Reddy VD, Chakravarthula VN. Nephro-protective activity of wheatgrass juice against alcohol-induced oxidative damage in rats. Toxicology Mechanisms and Methods. 2020;30(9):679-86.
• 23. Skoczylas Ł, Korus A, Tabaszewska M, Gędoś K, Szczepańska E. Evaluation of the quality of fresh and frozen wheatgrass juices depending on the time of grass harvest. Journal of Food Processing and Preservation. 2018;42(1):e13401.
• 24. Thakur N, Dhaliwal HS, Sharma V. Qualitative and Quantitative RP-HPLC-PDA Method of Analysis of Polyphenols in Lyophilized Wheat Seedling Juice Powder. International Journal on Emerging Technologies 11(2): 36-43
• 25. Shakya G, Balasubramanian S, Rajagopalan R. Methanol extract of wheatgrass induces G1 cell cycle arrest in a p53-dependent manner and down regulates the expression of cyclin D1 in human laryngeal cancer cells-an in vitro and in silico approach. Pharmacognosy Magazine. 2015;11(42):139.
• 26. Fjällskog MLH, Granberg DP, Welin SL, Eriksson C, Öberg KE, Janson ET, et al. Treatment with cisplatin and etoposide in patients with neuroendocrine tumors. Cancer: Interdisciplinary International Journal of the American Cancer Society. 2001;92(5):1101-7.
• 27. Maranchie JK. Silencing of Nox4 enhances cisplatin chemosensitivity of renal cell carcinoma. The Journal of Urology. 2008;179(4S):37-37.
• 28. Mirmalek SA, Azizi MA, Jangholi E, Yadollah-Damavandi S, Javidi MA, Parsa Y, et al. Cytotoxic and apoptogenic effect of hypericin, the bioactive component of Hypericum perforatum on the MCF-7 human breast cancer cell line. Cancer cell international. 2015;16(1):1-9.
• 29. Gibb RK, Taylor DD, Wan T, O'Connor DM, Doering DL, Gerçel- Taylor Ç. Apoptosis as a measure of chemosensitivity to cisplatin and taxol therapy in ovarian cancer cell lines. Gynecologic oncology. 1997;65(1):13-22.
• 30. Hosoyamada M, Obinata M, Suzuki M, Endou H. Cisplatin-induced toxicity in immortalized renal cell lines established from transgenic mice harboring temperature sensitive SV40 large T-antigen gene. Archives of toxicology. 1996;70(5):284-92.
• 31. Hazman Ö, Evin H, Bozkurt MF, Ciğerci İH. Hazman, Ömer, et al. "Two faces of arbutin in hepatocellular carcinoma (HepG2) cells: Anticarcinogenic effect in high concentration and protective effect against cisplatin toxicity through its antioxidant and anti-inflammatory activity in low concentration. Biologia. 2022; 77 :225–239
• 32. Shu X, Fan C, Long B, Zhou X, Wang Y. The anti-cancer effects of cisplatin on hepatic cancer are associated with modulation of miRNA-21 and miRNA-122 expression. Eur Rev Med Pharmacol Sci. 2016;20(21):4459-65.
• 33. Parihar A, Parihar MS, Ghafourifar P. Significance of mitochondrial calcium and nitric oxide for apoptosis of human breast cancer cells induced by tamoxifen and etoposide. International journal of molecular medicine. 2008;21(3):317-24.
• 34. Ardeshiry LA, Rezaie TM, Mortazavi SA, Barzegar M, Moghadamnia SH, Rezaee MB. Study of anti cancer property of Scrophularia striata extract on the human astrocytoma cell line (1321). 2010; 9 (4): 403-410.
• 35. Mozaffari F, Lindemalm C, Choudhury A, Granstam-Björneklett H, Helander I, Lekander M, et al. NK-cell and T-cell functions in patients with breast cancer: effects of surgery and adjuvant chemo-and radiotherapy. British journal of cancer. 2007;97(1):105-11.
• 36. Wijayahadi N, Haron M, Stanslas J, Yusuf Z. Changes in cellular immunity during chemotherapy for primary breast cancer with anthracycline regimens. Journal of chemotherapy. 2007;19(6):716-23.
• 37. Murta EFC, de Andrade JM, Falcio RP, Bighetti S. Lymphocyte subpopulations in patients with advanced breast cancer submitted to neoadjuvant chemotherapy. Tumori Journal. 2000;86(5):403-7.
• 38. Sabbioni ME, Bernhard J, Siegrist H-P, Schmitz S-FH, Gertsch MC, Thürlimann B, et al. Does subjective burden of early breast cancer and its treatment affect immune measures during adjuvant therapy? Breast cancer research and treatment. 2004;87(1):75-86.
• 39. Onyema OO, Decoster L, Njemini R, Forti LN, Bautmans I, De Waele M, et al. Chemotherapy-induced changes and immunosenescence of CD8+ T-cells in patients with breast cancer. Anticancer research. 2015;35(3):1481-9.
• 40. Kumar A, Fillmore HL, Kadian R, Broaddus WC, Tye GW, Van Meter TE. The Alkylphospholipid Perifosine Induces Apoptosis and p21-Mediated Cell Cycle Arrest in MedulloblastomaPerifosine Induces Apoptosis and Mitotic Arrest. Molecular Cancer Research. 2009;7(11):1813-21.
• 41. Suman G, Jamil K. Application of human lymphocytes for evaluating toxicity of anti-cancer drugs. Int J Pharmacol. 2006;2(4):374-81.
• 42. Atha DH, Coskun E, Erdem O, Tona A, Reipa V, Nelson BC. Genotoxic effects of etoposide, bleomycin, and ethyl methanesulfonate on cultured CHO cells: Analysis by GC-MS/MS and comet assay. Journal of nucleic acids. 2020;2020:1-10