Research Article


DOI :10.26650/JARHS2023-1175367   IUP :10.26650/JARHS2023-1175367    Full Text (PDF)

INVESTIGATION OF THE EFFECTS OF CHLORPYRIFOS AND TANNIC ACID APPLIED TO HEK 293 CELLS IN DIFFERENT DOSES AND TIMES ON CELL VITALITY

Sevil ErolBahar Öztürk KurtBircan DinçSemra Özdemir

Objective: Chlorpyrifos, an organophosphate pesticide, is a widely used insecticide and shows its effect with poisoning by inhibiting the activity of acetylcholinesterase, causing acetylcholine accumulation in neuromuscular junctions, heart muscle, smooth muscle and central nervous system. Pesticides are risk factors for cancer, neurodegenerative diseases, neurotoxicity, reproductive dysfunction and deterioration of many organ functions. Tannic acid is a polyphenol used in traditional and modern medicine with its many properties such as antioxidant, anti-inflammatory and anticarcinogenic. Our study was planned to investigate the possible effects of the application of chlorpyrifos and tannic acid at different doses and durations on the cell viability of the human embryonic kidney cell HEK293 cell line. Materials and Method: The study was carried out on 3 groups as control group, chlorpyrifos group and tannic acid group. MTT and xCELLigence analysis methods were used to determine at which doses and at what time chlorpyrifos and tannic acid were more effective on cell viability in HEK 293 cells. For this purpose, incubation was carried out by adding 10, 30, 60 and 90 μg/ml chlorpyrifos and 5, 10, 15, 20 and 25 μM tannic acid to the cell culture medium. Results: It was observed that the cell viability of the 30 μg/ml, 60 μg/ml and 90 μg/ml chlorpyrifos groups decreased significantly over the 24, 48 and 72 hours compared to the control group. The IC50 value of chlorpyrifos was found to be 79.94 μg/ml at 24-hour exposure. It was determined that the absorbance value of the group in which 25 μM of tannic acid was applied decreased significantly compared to the other study groups (p<0.001). Conclusion: It has been observed that chlorpyrifos is effective on cell viability especially at high doses and decreases viability more. It has been determined that the effective dose of tannic acid in increasing cell viability is 10µM and it is effective in reducing viability at high doses.

DOI :10.26650/JARHS2023-1175367   IUP :10.26650/JARHS2023-1175367    Full Text (PDF)

HEK 293 HÜCRELERİNE FARKLI DOZ VE SÜRELERDE UYGULANAN KLORPİRİFOS VE TANNİK ASİTİN HÜCRE CANLILIĞI ÜZERİNE ETKİLERİNİN ARAŞTIRILMASI

Sevil ErolBahar Öztürk KurtBircan DinçSemra Özdemir

Amaç: Organofosfatlı bir pestisit olan klorpirifos, yaygın olarak kullanılan bir böcek öldürücüdür ve etkisini asetilkolinesterazın etkinliğini engelleyerek, kalp kasında, düz kasta, merkezi sinir sisteminde asetilkolin birikimine neden olarak zehirlenme ile gösterir. Pestisitler kanserin, nörodejeneratif hastalıkların, üreme bozukluğunun ve birçok organ fonksiyonunun bozulmasında risk faktörüdürler. Tannik asit(TA) antioksidan, antiinflamatuar, antikanserojen özellikleri ile geleneksel ve modern tıpta kullanılan bir polifenoldür. Çalışmamız insan embriyonik böbrek hücresi olan HEK 293 hücre hattına klorpirifos ve tannik asitin farklı doz ve sürelerde uygulanmasının hücre canlılığı üzerine olası etkilerini araştırmak üzere planlanmıştır. Gereç ve Yöntem: Çalışma, kontrol, klorpirifos ve tannik asit grubu olmak üzere 3 grup ile yapılmıştır. HEK 293 hücrelerinde klorpirifos ve tannik asitin hücre canlılığı üzerine hangi dozlarda ve hangi sürelerde daha etkin olduğunu belirlemek amacıyla MTT [3-(4,5-dimetiltiyazol-2-il)-2,5- difeniltetrazolyum-bromid] ve xCELLigence analiz yöntemleri kullanılmıştır. Bu amaçla hücre kültürü ortamına 10, 30, 60 ve 90 μg/ml klorpirifos ve 5, 10, 15, 20 ve 25 μM tannik asit eklenerek inkübasyon gerçekleştirilmiş ve hücre canlılığı analizleri yapılmıştır. Bulgular: 30 μg/ml, 60 μg/ml ve 90 μg/ml klorpirifos uygulanan gruplarda 24, 48 ve 72 saatlik sürede hücre canlılığının kontrol grubuna göre anlamlı olarak azaldığı görülmüştür. Klorpirifosa ait IC50 (%50 baskılayıcı konsantrasyon) değeri 24 saatlik maruziyette 79,94 μg/ml bulunmuştur. Tannik asitin 25 μM uygulandığı grubun absorbans değerininin diğer gruplara göre anlamlı olarak azaldığı tespit edilmiştir (p<0.001). Sonuç: Klorpirifosun hücre canlılığı üzerinde özellikle yüksek dozlarda etkili olduğu ve canlılığı daha çok azalttığı görülmüştür. Tannik asitin hücre canlılığını artırmada etkili dozunun 10µM olduğu ve yüksek dozlarda canlılığı azaltma yönünde etkili olduğu tespit edilmiştir. 


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References

  • 1. Kurutaş EB, Kılınç M. Pestisitlerin biyolojik sistemler üzerine etkisi. Arşiv Kaynak Tarama Dergisi 2003;12(3):215-28. google scholar
  • 2. Vural N. Toksikoloji. Ankara Üniversitesi Eczacılık Fakültesi Yayınları. Ankara. 2005.p.344-55. google scholar
  • 3. Sağmanlıgil H. Tavuklarda ivermektin ve klorprifosun toksikolojik etkileşim özellikleri üzerinde deneysel araştırmala., YYÜ Vet Fak Derg 1994;5(1-2):65-84. google scholar
  • 4. Türkmen R, Özdemir M. Klorprifos-Etil uygulanan diyabetli ratlarda likopenin antioksidan ve hipoglisemik etkilerinin araştırılması. Kocatepe Vet J 2015;8(1):1-17. google scholar
  • 5. Mercan S, Eren B, Dinç N. Pestisit klorprifosun rat (wistar albino) karaciğerinde oluşturduğu hasar üzerine kurkuminin antioksidan etkisinin incelenmesi. Anadolu J Agr Sci 2017;32(1):132-38. google scholar
  • 6. Kayhan FE, Kaymak G, Akbulut C, Ertuğ ND. 2,4-D (Diklorofenoksiasetik Asit)’in zebra balığı (Danio rerio Hamilton, 1822) solungaçlarında antioksidan enzimler ve lipid peroksidasyon seviyesi üzerine akut etkilerinin belirlenmesi. Trakya Univ J Nat Sci 2017;18(2):143-48. google scholar
  • 7. Baldwin A, Booth B. Biomedical applications of tannic acid. J Biomaterials Appl 2022;36(8):1503-23. google scholar
  • 8. Wilson WC, Macgregor AR, Stewart CP. The clinical course and pathology of burns and scalds under modern methods of treatment. Br J Surg 1938;25(100):826-65. google scholar
  • 9. Perumal P, Mhlanga P, Somboro MA, Amoako DG, Khumalo HM, Khan RM. Cytoproliferative and anti-oxidant effects ınduced by tannic acid in human embryonic kidney (Hek-293) cells. Biomolecules 2019;9(12):767-84. google scholar
  • 10. Croft K. Dietary polyphenols: Antioxidants or not? Arch Biochem Biophys 2016;595:120-24. google scholar
  • 11. Wang TY, Li Q, Bi KS. Bioactive flavonoids ın medicinal plants: Structure, activity and biological fate. Asian J Pharm Sci 2018;13(1):12-23. google scholar
  • 12. Zhang PY. Polyphenols in health and disease. Cell Biochem Biophys 2015;73:649-64. google scholar
  • 13. Somboro AM, Osei Sekyere J, Amoako DG, Kumalo HM, Khan R, Bester LA, et al. In vitro potentiation of carbapenems with tannic acid against carbapenemase-producing enterobacteriaceae: Exploring natural products as potential carbapenemase inhibitors. J Appl Microbiol 2019;126(2):452-67. google scholar
  • 14. Badhani B, Sharma N, Kakkar R. Gallic acid: A versatile antioxidant with promising therapeutic and industrial applications. RSC Adv 2015;5:27540-57. google scholar
  • 15. Myint KB, Sing LC, Wei Z. Tannic acid as phytochemical potentiator for antibiotic resistance adaptation. APCBEE Procedia 2013;7:175-81. google scholar
  • 16. Sieniawska E, Baj T. Tannins. In: Simone B, Rupika D, editors. Pharmacognosy. Elsevier London UK 2017, pp.199-232. google scholar
  • 17. Gülçin İ, Huyut Z, Elmastaş M, Aboul-Enein HY. Radical scavenging and antioxidant activity of tannic acid. Arab J Chem 2010;3(1):43-53. google scholar
  • 18. Greenwell M and Rahman PK. Medicinal plants: Their use in anticancer treatment. Int J Pharm Sci Res 2015;6(10):4103-12. google scholar
  • 19. Khan HY, Hadi SM, Mohammad RM, Azmi AS. Prooxidant anticancer activity of plant-derived polyphenolic compounds: An underappreciated phenomenon. Functional foods in cancer prevention and therapy. London: Academic Press 2020, pp.221-36. google scholar
  • 20. Ullah MF, Khan HY, Zubair H, Shamim U, Hadi SM. The antioxidant ascorbic acid mobilizes nuclear copper leading to a prooxidant breakage of cellular DNA: implications for chemotherapeutic action against cancer. Cancer Chemother Pharmacol 2011;67(1): 103-10. google scholar
  • 21. Farhan M, Oves M, Chibber S, Hadi SM, Ahmad A. Mobilization of nuclear copper by green tea polyphenol epicatechin-3-gallate and subsequent prooxidant breakage of cellular DNA: implications for cancer chemotherapy. Int J Mol Sci 2017;18(1):34-48. google scholar
  • 22. Tatlı O. İnorganik veya organik çinko kapsayan etlik piliç rasyonlarına tannik asit katılmasının performans ve sindirilebilirlik üzerine etkisi. Adnan Menderes Üniv. 2015. google scholar
  • 23. Aşkın M. Ratlarda midpalatal sutur osteotomisi ile uygulanan üst çene genişletmesi sonrası kemik oluşumuna tannik asitin etkisinin değerlendirilmesi. Atatürk Üniv. 2019. google scholar
  • 24. Çalışlar S. Tanenlerin kanatlı hayvan beslemede etkileri. KSÜ Tarım ve Doğa Derg 2018;21(4):615-23. google scholar
  • 25. Taşpınar E. Klorprifos uygulanan HEK 293 hücrelerinde antioksidan kullanımının apoptoz ve oksidan sistem üzerine etkisinin incelenmesi. İstanbul Üniv. 2019. google scholar
  • 26. Özsobacı N. Elektromanyetik alan uygulamasının (Wİ-Fİ 2.4 GHZ) insan embriyonik böbrek hücrelerinde (HEK293) apoptoz ve oksidatif stres değerleri üzerine selenyum ve çinkonun etkisi. İstanbul Üniv. 2016. google scholar
  • 27. Ergün D. Hipoksi ve hiperkapninin insan embriyonik böbrek hücre membranında (HEK293) TRPM2 kanal aktivitesine etkisinin patch clamp tekniği ve enzimatik ölçümler ile araştırılması. İstanbul Üniv. 2018. google scholar
  • 28. Chu X, Guo Y, Xu B, Li W, Lin Y, Sun X, et al. Effects of tannic acid, green tea and red wine on hERG channels expressed in HEK293 cells. Plos One 2015;10(12):1-13. google scholar
  • 29. xCELLigence Real Time Cell Analyzer. Genomax Technologies. URL: https://www.genomaxtech.com/index.php/popular-articles/147-xCellingence. google scholar
  • 30. Ke N, Wang X, Xu X, Abbasi YA. The xCELLigence system for realtime and label-free monitoring of cell viability. Methods Mol Biol 2011;740:33-43. google scholar
  • 31. Demirdöğen B. Organofosfatlı pestisit zehirlenmeleri ve serum paraoksonaz 1 (PON1) enziminin organofosfat metabolizmasındaki rolü. Türk Hijyen ve Deneysel Biyoloji Derg 2010;67(2):97-112. google scholar
  • 32. Sun L, Zhang S, Guo W, He W, Qian Y, Qu G, et al. Sublethal exposure of organophosphate pesticide chlorpyrifos alters cellular iron metabolism in hepatocytes and macrophages. Int J Mol Med 2014;34(5):1395-400. google scholar
  • 33. Poliakova J, Pistl J, Kovalkovicova N, Csank T, Kocisova A, Legath J. Use of cultured cells of mammal and insect origin to assess cytotoxic effects of the pesticide chlorpyrifos. Pol J Environ Stud 2012;21(4):1001-06. google scholar
  • 34. Mitra I, Saha A and Roy K. Exploring quantitative structureactivity relationship studies of antioxidant phenolic compounds obtained from traditional Chinese medicinal plants. Mol Simulation 2010;36(13):1067-79. google scholar
  • 35. Kaczmarek B. Tannic acid with antiviral and antibacterial activity as a promising component of biomaterials—A minireview. Materials 2020;13(14):3224. google scholar
  • 36. Saraiva MA, Castro RH, Cordeiro RP, Sobrinho TP, Castro VA, Amorim EC, et al. In vitro evaluation of antioxidant, antimicrobial and toxicity properties of extracts of Schinopsis brasiliensis Engl. (Anacardiaceae). Afr J Pharm Pharmacol 2011;5(14):1724-31. google scholar

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APA

Erol, S., Öztürk Kurt, B., Dinç, B., & Özdemir, S. (2023). INVESTIGATION OF THE EFFECTS OF CHLORPYRIFOS AND TANNIC ACID APPLIED TO HEK 293 CELLS IN DIFFERENT DOSES AND TIMES ON CELL VITALITY. Journal of Advanced Research in Health Sciences, 6(1), 56-63. https://doi.org/10.26650/JARHS2023-1175367


AMA

Erol S, Öztürk Kurt B, Dinç B, Özdemir S. INVESTIGATION OF THE EFFECTS OF CHLORPYRIFOS AND TANNIC ACID APPLIED TO HEK 293 CELLS IN DIFFERENT DOSES AND TIMES ON CELL VITALITY. Journal of Advanced Research in Health Sciences. 2023;6(1):56-63. https://doi.org/10.26650/JARHS2023-1175367


ABNT

Erol, S.; Öztürk Kurt, B.; Dinç, B.; Özdemir, S. INVESTIGATION OF THE EFFECTS OF CHLORPYRIFOS AND TANNIC ACID APPLIED TO HEK 293 CELLS IN DIFFERENT DOSES AND TIMES ON CELL VITALITY. Journal of Advanced Research in Health Sciences, [Publisher Location], v. 6, n. 1, p. 56-63, 2023.


Chicago: Author-Date Style

Erol, Sevil, and Bahar Öztürk Kurt and Bircan Dinç and Semra Özdemir. 2023. “INVESTIGATION OF THE EFFECTS OF CHLORPYRIFOS AND TANNIC ACID APPLIED TO HEK 293 CELLS IN DIFFERENT DOSES AND TIMES ON CELL VITALITY.” Journal of Advanced Research in Health Sciences 6, no. 1: 56-63. https://doi.org/10.26650/JARHS2023-1175367


Chicago: Humanities Style

Erol, Sevil, and Bahar Öztürk Kurt and Bircan Dinç and Semra Özdemir. INVESTIGATION OF THE EFFECTS OF CHLORPYRIFOS AND TANNIC ACID APPLIED TO HEK 293 CELLS IN DIFFERENT DOSES AND TIMES ON CELL VITALITY.” Journal of Advanced Research in Health Sciences 6, no. 1 (Jul. 2024): 56-63. https://doi.org/10.26650/JARHS2023-1175367


Harvard: Australian Style

Erol, S & Öztürk Kurt, B & Dinç, B & Özdemir, S 2023, 'INVESTIGATION OF THE EFFECTS OF CHLORPYRIFOS AND TANNIC ACID APPLIED TO HEK 293 CELLS IN DIFFERENT DOSES AND TIMES ON CELL VITALITY', Journal of Advanced Research in Health Sciences, vol. 6, no. 1, pp. 56-63, viewed 13 Jul. 2024, https://doi.org/10.26650/JARHS2023-1175367


Harvard: Author-Date Style

Erol, S. and Öztürk Kurt, B. and Dinç, B. and Özdemir, S. (2023) ‘INVESTIGATION OF THE EFFECTS OF CHLORPYRIFOS AND TANNIC ACID APPLIED TO HEK 293 CELLS IN DIFFERENT DOSES AND TIMES ON CELL VITALITY’, Journal of Advanced Research in Health Sciences, 6(1), pp. 56-63. https://doi.org/10.26650/JARHS2023-1175367 (13 Jul. 2024).


MLA

Erol, Sevil, and Bahar Öztürk Kurt and Bircan Dinç and Semra Özdemir. INVESTIGATION OF THE EFFECTS OF CHLORPYRIFOS AND TANNIC ACID APPLIED TO HEK 293 CELLS IN DIFFERENT DOSES AND TIMES ON CELL VITALITY.” Journal of Advanced Research in Health Sciences, vol. 6, no. 1, 2023, pp. 56-63. [Database Container], https://doi.org/10.26650/JARHS2023-1175367


Vancouver

Erol S, Öztürk Kurt B, Dinç B, Özdemir S. INVESTIGATION OF THE EFFECTS OF CHLORPYRIFOS AND TANNIC ACID APPLIED TO HEK 293 CELLS IN DIFFERENT DOSES AND TIMES ON CELL VITALITY. Journal of Advanced Research in Health Sciences [Internet]. 13 Jul. 2024 [cited 13 Jul. 2024];6(1):56-63. Available from: https://doi.org/10.26650/JARHS2023-1175367 doi: 10.26650/JARHS2023-1175367


ISNAD

Erol, Sevil - Öztürk Kurt, Bahar - Dinç, Bircan - Özdemir, Semra. INVESTIGATION OF THE EFFECTS OF CHLORPYRIFOS AND TANNIC ACID APPLIED TO HEK 293 CELLS IN DIFFERENT DOSES AND TIMES ON CELL VITALITY”. Journal of Advanced Research in Health Sciences 6/1 (Jul. 2024): 56-63. https://doi.org/10.26650/JARHS2023-1175367



TIMELINE


Submitted14.09.2022
Accepted11.01.2023
Published Online09.02.2023

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