Investigation of Some Pesticides’ Effects on Activities of Glutathione Reductase and Glutathione S-Transferase Purified from Turkey Liver under in Vitro Conditions

Öz

Whereas a very small amount of pesticides which are used for elimination of undesirable species

and for a more fertile agriculture reach the target organism, majority of pesticides reach to nontarget organisms. It is

foreseen by our team that the pesticides used for various purposes may negatively affect the glutathione mechanism

of the organisms. However, determination of effective dosage range is the objective of our study subjects. For this

reason, the in vitro effects of widely used pesticides (lambda-cyhalothrin, cypermethrin, chlorpyrifos, dichlorvos,

glyphosate isopropylamine) on the activities of glutathione reductase (GR) and glutathione S-transferase

(GST) which are two important enzymes of the glutathione system have been investigated in this study. It has

been determined that when lambda-cyhalothrin does not affect the GR enzyme purified from the turkey liver,

chlorpyrifos, glyphosate isopropylamine, dichlorvos and cypermethrin cause inhibition. When the effects of

pesticides on GST enzyme purified from turkey liver were investigated in in vitro conditions, it was determined

that all examined pesticides had inhibitory effects. In this context, the potential doses which can create a risk for

live life, of pesticides that are commonly used, have been identified.

Anahtar Kelimeler

• Glutathione S-transferase,glutathione reductase; pesticides

Hindi Karaciğerinden Saflaştırılan Glutatyon Redüktaz ve Glutatyon S-Transferaz Aktiviteleri Üzerine Bazı Pestisitlerin Etkilerinin in Vitro Şartlarda İncelenmesi

Öz

İstenmeyen türlerin ortadan kaldırılması ve daha verimli bir tarım için kulanılan pestisitlerin, çok küçük

bir kısmı hedef organizmaya ulaşırken oldukça büyük bir kısmı da hedef olmayan organizmalara ulaşmaktadır.

Çeşitli amaçlarla kullanılan pestisitlerin söz konusu organizmaların glutatyon mekanizmasını olumsuz şekilde

etkileyebileceği ekibimiz tarafından öngörülmüştür. Ancak etkili dozaj aralığının tespit edilmesi çalışma konumuzun

amacını oluşturmaktadır. Bu nedenle belirtilen çalışmada glutatyon sisteminin iki önemli enzimi olan glutatyon

redüktaz (GR) ve glutatyon S-transferazın (GST) aktiviteleri üzerine oldukça yaygın kullanılan pestisitlerin

(lambda-sihalotrin, sipermetrin, klorpirifos, diklorvos, glifosat izopropilamin) in vitro etkileri araştırılmıştır. Hindi

karaciğerinden saflaştırılan GR enzim aktivitesi üzerine lambda-sihalotrin’nin etki etmediği, klorpirifos, glifosat

izopropilaminin, diklorvos ve sipermetrinin ise inhibisyona sebep olduğu tespit edilmiştir. Hindi karaciğerinden

saflaştırılan GST enzim aktivitesi üzerine söz konusu pestisitlerin etkileri in vitro şartlarda araştırıldığında ise

çalışma kapsamında incelenen tüm pestisitlerin inhibisyon etkisine sahip olduğu belirlenmiştir. Bu kapsamda

yaygın olarak kullanılan zirai ilaçların canlı yaşam için risk oluşturabilecek potansiyel dozajları tespit edilmiştir.

Anahtar Kelimeler

• Glutatyon redüktaz,glutatyon S-transferaz,pestisit

Kaynakça

1. Akkemik E, Taser P, Bayindir A, Budak H, Ciftci M, 2012. Purification and characterization of glutathione S-transferase from turkey liver and inhibition effects of some metal ions on enzyme activity. Environmental Toxicology and Pharmacology, 34: 888-894.
2. Aksoy M, Özaslan MS, Küfrevioğlu OI, 2016. Purification of glutathione S-transferase from Van Lake fish (Chalcalburnus tarichii Pallas) muscle and investigation of some metal ions effect on enzyme activity J Enzyme Inhib Med Chem, 31(4): 546–550.
3. Alaa AM, Manal HA, Manal EA, Halwagy E, Mostafa M, Afify M, 2011. A study on oxidative stress biomarkers and immunomodulatory effects of pesticides in pesticide-sprayers. Egyptian Journal of Forensic Sciences, 1: 93–98.
4. Al-Ghais, SM, Ali B, 1999. Inhibition of Glutathione S-Transferase Catalyzed Xenobiotic Detoxication by Organotin Compounds in Tropical Marine Fish Tissues Bull. Environ. Contam. Toxicol. 62:207-213.
5. Armstrong RN, 1997. Structure, catalytic mechanism, and evolution of the glutathione transferases. Chemical Research in Toxicology, 10(1): 2-18.
6. Bhuvarahamurthy V, Balasubramanian N, Govindasamy S, 1979. Effect of radiotherapy and chemoradiotherapy on circulating antioxidant system of human uterine cervical carcinoma. Molecular and Cellular Biochemistry, 158(1): 17-23.
7. Burçak AA, 2012. İlaç, Alet ve Toksikoloji Araştırmaları Çalışma Grubu, Bitki Sağlığı Araştırmaları Daire Başkanlığı T.C. Tarım Gıda ve Hayvancılık Bakanlığı. http://www.tarim.gov.tr/TAGEM/Belgeler/SUNULAR
8. Carlberg I, Mannervik B, 1975. Purification and characterization of the flavoenzyme glutathione reductase from rat liver. The Journal of Biological Chemistry, 250: 5475- 5480.

9. Ceyhun SB, Sentürk M, Erdoğan O, Küfrevioğlu ÖI, 2010. In vitro and in vivo effects of some pesticides on carbonic anhydrase enzyme from rainbow trout (Oncorhynchus mykiss) gills Pesticide Biochemistry and Physiology 97, 177–181.
10. Ceylan S, 1977. Klorlu hidrokarbon rezidülerinin süt, tereyağı, peynir ve iç yağlannda kramotografik yöntemlerle araştınlması. Ankara Üniversitesi Veteriner Fakültesi Dergisi, 24(2): 296-318.
11. Dilek ON, 2003. Serbest Radikaller ve Cerrahi. Serbest Radiakaller ve Antioksidanlar Araştırma Derneği III.Ulusal Kongresi: 23-30 Mart,Afyon.
12. Douglas, K.T.,1987. Mechanizm of glutathione-dependent enzymes., Meister, A.,(ed). Avdan. Enzymol., Johm Wiley and Sons ınc., New York. 59, 103-167.
13. Durmusoglu E, 2003. Market basket monitoring of some organophoshorus pesticides on apple and strawberry in Izmir province, Turkey. Archiv für Lebensmittelhygiene. 54(1): 16-19.
14. Fridovich I, 1976. In free radical in biology. In: Pryor W A (Ed.), Academic, 1. Edt., New York, USA. 239-271.
15. Gultekin F, Ozturk M, Akdogan M, 2000. The effect of organophosphate insecticide chlorpyrifos-ethyl on lipid peroxidation and antioxidant enzymes (in vitro). Archives of Toxicology, 74: pp 533–538.
16. Güller U, Taşer P, Çiftci M, Küfrevioğlu Öİ, 2014. Purification of Glutathione S-Transferase From Bonito (Sarda Sarda) Liver And Investigation of Metal Ions Effects on Enzyme Activity. Hacettepe J. Biol. &Chem., 42 (3),435-442.
17. Habig WH, Pabst MJ, Jakoby WB, 1974. Glutathione S- transferases. The first enzymmatic step in mercapturic acid formation. Journal of Biological Chemistry, 246: 7130-7139.
18. Halliwell B, 2007. Biochemistry of oxidative stress. Biochemical Society Transactions, 35(5): 1147-1150.
19. Hayes JD, Pulford DJ, 1995. The glutathione S-transferase supergene family: regulation of GST and the contribution of the isoenzymes to cancer chemoprotection and drug resistance. Critical Reviews in Biochemistry and Molecular Biology, 30: 445-600.
20. Henning JC, Davis AC, Robinson WB, 1954. Flavor and color evaluation of canning crops grown in soil treated with insecticides. Journal of Food Science and Technology, 8: 227. http://www.resmigazete.gov.tr/eskiler/2013/03/20130307-7.htm
21. Keha EE, Küfrevioglu Öİ, 2012. Biyokimya. Aktif Yayınevi. 9. Baskı, Erzurum, Türkiye. 643 s.
22. Kovesdy CP, Kalantar-Zadeh K, 2009. Review article: Biomarkers of clinical outcomes in advanced chronic kidney disease. Nephrology (Carlton), 4(4): 408-415.
23. Lineweaver H, Burk D, 1934. The determination of enzyme dissocation constants. Journal of the American Chemical Society, 57: 685.
24. Özaslan MS, Demir Y, Küfrevioğlu OI, 1 Çiftci M, 2017. Some metals inhibit the glutathione S-transferase from Van Lake fish gills J BiochemMol Toxicol.;31:e21967.
25. Romero FJ, Bosch-Morell F, Romero MJ, Jareno EJ, Romero B, Marin N, Roma J, 1998. Lipid peroxidation products and antioxidants in human disease. Environmental Health Perspectives, 106(5): 1229-1234.
26. Sabuncuoğlu S, Özgüneş H, 2011. Kemoterapi, Serbest Radikaller ve Oksidatif Stres. Hacettepe Üniversitesi Eczacılık Fakültesi Dergisi, 31(2): 137-150.
27. Sentürk M, Ceyhun SB, Erdoğan O, Küfrevioğlu ÖI, 2009. In vitro and in vivo effects of some pesticides on glucose-6-phosphate dehydrogenase enzyme activity from rainbow trout (Oncorhynchus mykiss) erythrocytes Pesticide Biochemistry and Physiology 95 95–99.
28. Singh M, Sandhir R, Kiran R, 2006. Erythrocyte antioxidant enzymes in toxicological evaluation of commonly used organophosphate pesticides, Indian journal of Experimental Biology, 44; 580-583.
29. Taşer P, Ciftci M, 2012. Purification and characterization of glutathione reductase from turkey liver. Turkish Journal of Veterinary and Animal Sciences, 36(5): 546-553.
30. Tekman B, Ozdemir H, Senturk M, Ciftci M, 2008. Purification and characterization of glutathione reductase from rainbow trout (Oncorhynchus mykiss) liver and inhibition eff ects of metal ions on enzyme activity. Com. Biochem. Physiol. C; 148: 117-121.
31. Tiryaki O, Canhilal R, Horuz S, 2010. The use of pesticides and their risks. Erciyes Üniversitesi Fen Bilimleri Enstitüsü Dergisi, 26(2): 154-169.
32. Wildburger R, Mrakovcic L, Stroser M, Andrisic L, Borovic SS, Zarkovic K, Zarkovic N, 2009. Lipid peroxidation and age-associated diseases-cause or consequence? Review Citation. Türkiye Klinikleri Tıp Bilimleri Dergisi, 29(1): 189-193.
33. Yıldız M, Gürkan O, Turgut C, Kaya Ü, Ünal G, 2005. Tarımsal Savaşımda Kullanılan Pestisitlerin Yol Açtığı Çevre Sorunları VI. Türkiye Ziraat Mühendisliği Teknik Kongresi. TMMOB Ziraat Mühendisleri Odası, Ankara.