EFFECT OF THIACLOPRID ON DNA DAMAGE IN HUMAN LYMPHOCYTES

Öz

Objective Thiacloprid is a neonicotinoid insecticide and the number of studies on its effect on DNA damage is very limited. The aim of this study was to evaluate the effect of Thiacloprid on human lymphocyte DNA with different doses and exposure times. Material and Methods 15 mL of blood was drawn from each of 4 male and 4 female volunteers, and each was used separately for the groups included in the study. Thiacloprid was Deklarasyoadministered in 3 different doses as 100, 300 and 500 μM, and at 3 different times, 1, 2 and 4 hours for each dose. Internal positive and negative control groups, which created separately for each incubation hour, were used. The comet assay was used for the detection of DNA damage and the tail DNA percentage parameter was chosen as measure of damage. Results All Thiacloprid applications caused an increase in DNA damage. However, the increase in groups 7, 10 and 13, which incubated with the 100 μM Thiacloprid, was not statistically significant compared to the internal control groups (p>0.05). Groups 8, 9, 11, 12, 14, and 15, which incubated with 300 or 500 μM Thiacloprid for 1, 2, or 4 h, showed a statistically significant increase in DNA damage compared to the internal control groups (p<0.05). Conclusion Increases in DNA damage were dose-related rather than incubation time. DNA damage was appeared to increase as the Thiacloprid dose increases. Although much more extensive researches are required before making definite conclusions, the data, which obtained from previous researches and the result of our research, reveal that Thiacloprid is genotoxic.

Anahtar Kelimeler

• Comet assay
• Neonicotinoid
• Pesticide
• Thiacloprid

THİACLOPRİD’İN İNSAN LENFOSİTLERİNDE DNA HASARINA ETKİSİ

Öz

Amaç Thiacloprid neonikotinoid sınıfı bir insektisittir ve DNA hasarına etkisi hakkında yapılan çalışma sayısı oldukça sınırlıdır. Bu araştırma ile Thiacloprid’in insan lenfosit DNA’sı üzerindeki etkisinin farklı dozlar ve farklı maruziyet süreleri için değerlendirilmesi amaçlanmıştır. Gereç ve Yöntem 4 erkek ve 4 kadın gönüllünün her birinden 15 mL kan alınmış ve çalışmada yer alan gruplar için tüm katılımcıların kanları ayrı ayrı kullanılmıştır. Thiacloprid, 100, 300 ve 500 μM olmak üzere 3 farklı dozda ve her bir doz için 1, 2 ve 4 saat olmak üzere 3 farklı zamanda uygulanmıştır. Her inkübasyon saati için ayrı ayrı oluşturulan internal pozitif ve negatif kontrol grupları karşılaştırma için kullanılmıştır. DNA hasarının tespiti için comet metodu kullanılmış ve kuyruk DNA yüzdesi parametresi hasarın göstergesi olarak seçilmiştir. Bulgular Tüm Thiacloprid uygulamaları DNA hasarında artışa sebep olmuştur. Ancak en düşük doz olan 100 μM Thiacloprid ile inkübasyon uygulanan gruplar 7, 10 ve 13’de meydana gelen artış, internal kontrol gruplarına kıyasla istatistiksel olarak anlamlı bulunmamıştır (p>0,05). 300 veya 500 μM Thiacloprid ile 1, 2 veya 4 s inkübasyon uygulanan gruplar 8, 9, 11, 12, 14 ve 15’de, internal kontrol gruplarına kıyasla DNA hasarında anlamlı artış görülmüştür (p<0,05). Sonuç DNA hasarında meydana gelen artışlar inkübasyon süresinden ziyade doza bağlı olarak meydana gelmiştir. İnkübasyon süresi ne olursa olsun Thiacloprid dozu arttıkça DNA hasarında artış meydana geldiği görülmektedir. Kesin yargılara varmadan önce çok daha kapsamlı araştırmalar yapmak gerekiyorsa da, literatür taraması sonucunda elde edilen veriler ve araştırmamızın sonucu Thiacloprid’in genotoksik olduğunu ortaya koymaktadır.

Anahtar Kelimeler

• Comet metodu
• Neonikotinoid
• Pestisit
• Thiacloprid

Kaynakça

1. 1. Sanborn M, Kerr K, Sanin L, Cole D, Bassil K, Vakil C. Non-cancer health effects of pesticides Systematic review and implications for family doctors. Canadian Family Physician. 2007;53(10):1712-20.
2. 2. Araoud M. Biological Markers of human exposure to pesticides. Pesticides in the Modern World-Pests Control and Pesticides Exposure and Toxicity Assessment: IntechOpen; London, 2011.
3. 3. Toğay VA. Pestisit Thiacloprid'in Parkinson Hastalığı ile Bağlantısının Araştırılması [DoktoraTezi]. Isparta: Süleyman Demirel Üniversitesi; 2021.
4. 4. Altıkat A, Turan T, Torun FE, Bingül Z. Türkiye’de pestisit kullanımı ve çevreye olan etkileri. Atatürk Üniversitesi Ziraat Fakültesi Dergisi. 2009;40(2):87-92.
5. 5. Qian L, Cui F, Yang Y, Liu Y, Qi S, Wang C. Mechanisms of developmental toxicity in zebrafish embryos (Danio rerio) induced by boscalid. Science of The Total Environment. 2018;634:478- 87.
6. 6. Tomizawa M, Casida JE. Neonicotinoid insecticide toxicology: mechanisms of selective action. Annu Rev Pharmacol Toxicol. 2005;45:247-68.
7. 7. Morrissey CA, Mineau P, Devries JH, Sanchez-Bayo F, Liess M, Cavallaro MC, et al. Neonicotinoid contamination of global surface waters and associated risk to aquatic invertebrates: A review. Environment International. 2015;74:291-303.
8. 8. Toğay VA, Baş FY, Çelik DA, Özçelik N, Türel GY, Calapoğlu M, et al. Increased DNA Damage of Radiology Personnel Chronically Exposed to Low Levels of Ionizing Radiations. Süleyman Demirel Üniversitesi Sağlık Bilimleri Dergisi. 2020;11(2):212-6.

9. 9. Celik DA, Togay VA, Turel GY, Tuluceoglu EE, Kosar PA. DNA Damages of Widely Used Pesticides; A Comet Assay Report for Chlorothalonil and Glyphosate Potassium Salt. Fresenius Environ Bull. 2021;30(4 A):4170-6.
10. 10. Toğay VA, Yavuz Türel G, Aşcı Çelik D, Özgöçmen M, Evgen Tülüceoğlu E, Şen İ, et al. DNA damage effect of cyprodinil and thiacloprid in adult zebrafish gills. Environmental Science and Pollution Research. 2021;28(12):14482-7.
11. 11. Yavuz Türel G, Toğay VA, Aşcı Çelik D. Genotoxicity of thiacloprid in zebrafish liver. Archives of Environmental & Occupational Health. 2022:1-6. doi: 10.1080/19338244.2022.2118212.
12. 12. Aşci Çelik D, Toğay VA, Karabacak P. DNA damage assessment in pneumonia patients treated in the intensive care unit. Biologia (Bratisl). 2022;77(7):1909–1913.
13. 13. Karabacak P, Toğay VA, Çelik DA. Lymphocyte DNA damage in sepsis and septic-shock intensive-care patients: Damage is greater in non-intubated patients. Mutation Research/Genetic Toxicology and Environmental Mutagenesis. 2022;879:503516.
14. 14. OECD. Test No. 489: In Vivo Mammalian Alkaline Comet Assay. Paris: OECD Publishing; 2016.
15. 15. Gyori BM, Venkatachalam G, Thiagarajan P, Hsu D, Clement M-V. OpenComet: An automated tool for comet assay image analysis. Redox biology. 2014;2:457-65.
16. 16. IBM. SPSS Statistics for Windows, Version 20.0. Armonk, NY: IBM Corp.; 2012.
17. 17. Azmi MA, Naqvi S. Pesticide pollution, resistance and health hazards. In: Pesticides-The Impacts of Pesticides Exposure. Croatia: IntechOpen; 2011.
18. 18. Chen M, Tao L, McLean J, Lu C. Quantitative analysis of neonicotinoid insecticide residues in foods: implication for dietary exposures. Journal of agricultural and food chemistry. 2014;62(26):6082-90.
19. 19. Ospina M, Wong LY, Baker SE, Serafim AB, Morales-Agudelo P, Calafat AM. Exposure to neonicotinoid insecticides in the US general population: Data from the 2015-2016 national health and nutrition examination survey. Environmental Research. 2019;176: 108555. doi:10.1016/j.envres.2019.108555.
20. 20. Ueyama J, Nomura H, Kondo T, Saito I, Ito Y, Osaka A, et al. Biological monitoring method for urinary neonicotinoid insecticides using LC-MS/MS and its application to Japanese adults. Journal of occupational health. 2014;56(6):461-8.
21. 21. Ikenaka Y, Miyabara Y, Ichise T, Nakayama S, Nimako C, Ishizuka M, et al. Exposures of children to neonicotinoids in pine wilt disease control areas. Environmental toxicology and chemistry. 2019;38(1):71-9.
22. 22. Wang X, Anadón A, Wu Q, Qiao F, Ares I, Martínez-Larrañaga M-R, et al. Mechanism of neonicotinoid toxicity: Impact on oxidative stress and metabolism. Annu Rev Pharmacol Toxicol. 2018;58:471-507.
23. 23. Lerda D, Bistoni MB, Peralta N, Ychari S, Vazquez M, Bosio G. Fumonisins in foods from Cordoba (Argentina), presence and genotoxicity. Food and Chemical Toxicology. 2005;43(5):691-8.
24. 24. Çelik DA, Toğay VA, Türel GY, Özçelik N. Gıda Katkı Maddesi Olarak Kullanılan Sitrik Asit, Askorbik Asit ve Sodyum Sitratın İnsan Lenfosit Hücrelerinde Genotoksisitesinin Değerlendirilmesi. SDÜ Tıp Fakültesi Dergisi. 2022;29(3):486-92.
25. 25. Calderón-Segura ME, Gómez-Arroyo S, Villalobos-Pietrini R, Martínez-Valenzuela C, Carbajal-López Y, Calderón-Ezquerro MdC, et al. Evaluation of genotoxic and cytotoxic effects in human peripheral blood lymphocytes exposed in vitro to neonicotinoid insecticides news. Journal of toxicology. 2012;2012:612647. doi: 10.1155/2012/612647.
26. 26. Kocaman AY, Rencuzogullari E, Topaktas M. In vitro investigation of the genotoxic and cytotoxic effects of thiacloprid in cultured human peripheral blood lymphocytes. Environmental Toxicology. 2014;29(6):631-41.
27. 27. Galdíková M, Šiviková K, Holečková B, Dianovský J, Drážovská M, Schwarzbacherová V. The effect of thiacloprid formulation on DNA/chromosome damage and changes in GST activity in bovine peripheral lymphocytes. Journal of Environmental Science and Health, Part B. 2015;50(10):698-707.
28. 28. Şenyildiz M, Kilinc A, Ozden S. Investigation of the genotoxic and cytotoxic effects of widely used neonicotinoid insecticides in HepG2 and SH-SY5Y cells. Toxicology and industrial health. 2018;34(6):375-83.
29. 29. Sekeroglu V, Karabiyik A, Sekeroglu ZA. Co-exposure to deltamethrin and thiacloprid induces cytotoxicity and oxidative stress in human lung cells. Toxicology and Industrial Health. 2020;36(11):916-24.
30. 30. Sekeroglu V, Sekeroglu ZA, Kefelioglu H. Cytogenetic effects of commercial formulations of deltamethrin and/or thiacloprid on wistar rat bone marrow cells. Environmental Toxicology. 2013;28(9):524-31.
31. 31. Aydin B. Effects of thiacloprid, deltamethrin and their combination on oxidative stress in lymphoid organs, polymorphonuclear leukocytes and plasma of rats. Pesticide Biochemistry and Physiology. 2011;100(2):165-71.