Sıçan Eritrositlerinde Karbon Tetraklorürün(CCl4) ve Kuersetinin Bazı Metabolik Enzim Aktiviteleri Üzerine İn Vivo Etkisinin İncelenmesi

Abstract

Çalışmada; Sıçan eritrositlerinde karbon tetraklorür (CCl4) ve Kuersetinin(Ku)'in, Glukoz 6-fosfat dehidrogenaz (G6PD), 6-fosfoglukonat dehidrogenaz (6PGD), glutatyon redüktaz (GR) ve glutatyon S-transferaz (GST) gibi önemli metabolik enzim aktiviteleri üzerine in vivoetkilerinin incelenmesi hedeflendi.Deneysel aşamadasıçanlar 4 gruba ayrıldı. 1.Grup(Kontrol): Bu gruptaki ratlara vücut ağırlıklarına göre belirlenen dozda(1 mL/kg)saf zeytin yağı, 2.Grup(CCl4: 1.0 mL/kg (i.p.)(1:1), 3.Grup(Ku: 25 mg/kg (i.p),4.Grup(CCl4(1.0 ml/kg (i.p.)+ Ku (25 mg/kg (i.p.) enjekte edildi.Çalışma 3günsürdürüldü.Sonuçlar değerlendirildiğinde; G6PD (p˂0,01), 6PGD (p˂0,01) GR (p˂0,001) ve GST (p˃0,05) enzim aktiviteleri kontrol grubuna oranla CCl4grubunda azalmıştır. CCl4+Ku uygulanan gruplarda ise enzim aktivitelerinin CCl4grubuna oranla yüksekolduğu belirlenmiştir.Ku uygulanması, enzim aktivite değerinin yükselmesine neden olmuştur. BudaCCI4’ün neden olduğu inhibisyonun ortadan kalktığının göstergesi olarak kabul edilebilir. Sonuç olarak, CCl4uygulamasına bağlı olarak ortaya çıkan oksidatif hasarı önlemede Ku'nın etkili olabileceği düşünülmektedir.

Keywords

• CCl4
• ,Ku
• ,G6PD
• ,6PGD
• ,GR

Investigation of In Vivo Effects of Carbon Tetrachloride (CCl4) and Quercetin on Some Metabolic Enzyme Activities in Rat Erythrocyte

Abstract

In the study; the purpose was to investigate the in vivoimpact of carbon tetrachloride (CCl4) and quercetin (Qu) on activities of important metabolic enzymes such as Glucose 6-phosphate dehydrogenase (G6PD), 6-phosphogluconate dehydrogenase (6PGD), glutathione reductase (GR) and glutathione S-transferase (GST) in rat erythrocytes. At the experimental stage, rats were divided into 4 groups. 1.Group (Control): Pure olive oil at a dose determined according to their body weight (1mL/kg) was given to the rats in this group, 2.Group (CCl4: 1.0 mL/kg (ip)(1:1), 3.Group (Ku: 25) mg/kg (ip), 4.Group (CCl4(1.0 ml/kg (ip)+ Ku (25 mg/kg (ip) was injected. The study was continued for 3 days.The results revealed that the activities of; G6PD (p˂0.01), 6PGD (p˂0.01), GR(p˂0.001) and GST (p˃0.05) enzyme activities were decreased in the CCl4group compared to the control group. It was determined that enzyme activities were higher in CCl4+Qu applied groups compared to CCl4group.The application of Qu caused an increase in the enzyme activity value. This can be accepted as an indication that the inhibition caused by CCI4has disappeared. Consequently; It is thought that Qu may be effective in preventing oxidative damage due to CCl4administration.

Keywords

• Carbon tetrachloride
• Quercetin
• Glucose 6-phosphate dehydrogenase
• 6-phosphogluconate dehydrogenase
• Glutathione reductase
• Glutatyon S-transferaz

References

1. [1]Vural DF.Karbontetraklorür (CCl4) Uygulanmış Sıçanların Gastrocnemius Kasına Melatoninin Etkisi. Ankara Üniversitesi Fen Bilimleri Enstitüsü.Yüksek Lİsans Tezi.2015.
2. [2].Akkoyun-Bayramoğlu M, Akkoyun HT. Effect Of Carbon Tetrachlorıde (CCl4) And Ellagıc Acıd On Rat Erythrocyte G6PD, 6PGD, GR, GST AND TrxR Enzyme Actıvıtıes. Fresenius Environmental Bulletin.2019:28(9), 6779-6785.
3. [3].Akkoyun HT, Akkoyun-Bayramoğlu M. Effect Of Quercetın On Selected Mıcro Elements In Rat Lıver Tıssue Durıng Carbon Tetrachlorıde Exposure. Fresenius Environmental Bulletin.2019:28(9), 6757-6763.
4. [4].Al-Harbi NO, Imam F, Nadeem A, Al-Harbi MM, Iqbal M, Ahmad SF. Carbon tetrachloride-induced hepatotoxicity in rat is reversed by treatment with riboflavin. Int Immunopharmacol.2014:21(2):383-8.
5. [5].Akkuş İ. Serbest Radikaller ve Fizyopatolojik Etkileri. Mimoza Yayınları, Konya.1995.
6. [6].Murray RK, Mayes PA, Granner DK, and Rodwell VW. Harper’ın Biyokimyası. Barış Kitapevi.1993.
7. [7].Kayaalp O. Rasyonel Tedavi Yönünden Tıbbi Farmakoloji. 10. Baskı, Hacettepe Taş.2002.
8. [8].Kurt H, Başaran A , Musmul A. Sıçanlarda Karbon Tetraklorit (CCl4)’in Oluşturduğu Oksidatif Stresin Kateşin ile Önlenmesi. Kocatepe Tıp Dergisi.2004: 5(2),29-34.

9. [9].León OS, Menéndez S, Merino N, Castillo R, Sam S, Pérez L, Cruz E, Bocci V. Ozone oxidative preconditioning: a protection against cellular damage by free radicals. Mediators Inflamm.1998:7(4),289-94.
10. [10].Süha-Yalçin A, Koçak-Toker N, Uysal M, Aykaç G, Sivas A, Oz H. Stimulation of lipid peroxidation and impairment of glutathione-dependent defence system in the liver of rats repeatedly treated with carbon tetrachloride. J Appl Toxicol.(1986):6(4),303-6.
11. [11].Tekeli H,Bildik A. Karbon Tetraklorür İle Oluşturulan Karaciğer Hasarında Glutatyon (GSH) Ve Glutatyon S-Transferaz (GST) Aktivitesi Üzerine N-Asetil Sisteinin Etkisi. Balikesir Saglik Bil Derg.2016: 5(2).
12. [12].Akkoyun HT,Bengü AŞ,Ulucan A,Bayramoğlu-Akkoyun M,Arihan O.(2018). Protectıve Effect Of Ellagıc Acıd Agaınst Carbon Tetrachlorıde (CCl4) Induced Oxıdatıve Braın Injury In Rats. Fresenius Environmental Bulletin.2018: 27(5),3148-3155.
13. [13].Choi YJ, Kang JS, Park JH, Lee YJ, Choi JS, Kang YH. Polyphenolic flavonoids differ in their antiapoptotic efficacy in hydrogen peroxide treated human vascular endothelial cells. J Nutr. 2003:133,985–991.
14. [14].Pavanato A, Tunon MJ, Sanchez-Campos S, Marroni CA, Llesuy S, Gonzalez- Gallego J, Marroni N. Effects of quercetin on liver damage in rats with carbon tetrachloride-induced cirrhosis. Digestive Diseases and Sciences.2003:48(4), 824-829.
15. [15].Ergüzel ET. Quercetin (3, 3’, 4’, 5, 7- pentahidroksiflavon)’in Bakır (II) ve Çinko (II) Komplekslerin Kararlılık Sabitlerinin Tayini. Yüksek Lisans. İstanbul: Marmara Üniversitesi.2006.
16. [16].Emek H. Dönmez Burukoğlu D. Bayçu C, Musmul A. Erişkin erkek sıçanlarda karbon tetraklorür ile oluşturulan testis hasarı üzerine kuersetinin etkisi, Osmangazi Tıp Dergisi.2017:39(65-73).
17. [17].Cho JY, Kim IS, Jang YH, Kim AR, Lee SR. Protective effect of quercetin, a natural flavonoid against neuronal damage after transient global cerebral ischemia. Neurosci Lett.2006:404(3),330–335.
18. [18].Schültke E, Kamencic H, Zhao M, Tian GF, Baker AJ, Griebel RW, Juurlink BH. Neuroprotection following fluid percussion brain trauma: a pilot study using quercetin. J Neurotrauma.2005:22(12),1475–1484.
19. [19].Terao J. and Piskula M. Flavonoids as inhibitors of lipid peroxidation in membranes. In: Flavonoids in Health and Disease. Rice-Evans, C. and Packer, L., eds. Marcel Dekker: New York.1998: 277–93.
20. [20].Makni M, Chtourou Y, Garoui EM, BoudawaraT, Fetoui H. Carbon tetrachloride- induced nephrotoxicity and DNA damagein rats: Protective role of vanillin. Human and Experimental Toxicology.2012: 31, 844-852.
21. [21].Yuan Y, Ma S, Qi Y, Wei X, Cai H, Dong L, Guo Q. Quercetin inhibitedcadmium-induced autophagy in the mouse kidneyvia inhibition of oxidative stress. Journalof Toxicologic Pathology.2016: 29(4), 247-252.
22. [22].Bayramoğlu-Akkoyun M, Bengü AŞ, Temel Y, Akkoyun HT, Ekin S, Ciftci M. The effect of astaxanthin and cadmium on rat erythrocyte G6PD, 6PGD, GR, and TrxR enzymes activities in vivo and on rat erythrocyte 6PGD enzyme activity in vitro. J Biochem Mol Toxicol.2018:32(8),e22170.
23. [23].Beutler E. Redcell metabolism. Manual of biochemical methods Acad Pres London. 1971:12,68–70.
24. [24].Bayindir S, Temel Y, Ayna A, Ciftci M. The synthesis of N‐benzoylindoles as inhibitors of rat erythrocyte glucose‐6‐phosphate dehydrogenase and 6‐phosphogluconate dehydrogenase. Journal of biochemical and molecular toxicology.2018: 32(9), e22193.
25. [25].Bayindir S, Ayna A, Temel Y, Ciftci M. The synthesis of new oxindoles as analogs of natural product 3,3-bis(indolyl) oxindole and in vitro evaluation of the enzyme activity of G6PD and 6PGD. Turkish Journal of Chemistry.2018b:42(2),332-345.
26. [26].Carlberg I, Mannervik B.Purification and characterization of glutathione reductase from calf liver. Glutathione reductase assays. Methods Enzymol.1981:113,484–4.
27. [27].Habig WH, Pabst MJ, Jakoby WB. Glutathione S-transferases the first enzymatic step in mercapturic acid formation. Journal of Biological Chemistry.1974: 249(22), 7130-719.
28. [28].Temel Y, Ayna A, Hamdi Shafeeq I, Ciftci M. In vitro effects of some antibiotics on glucose-6-phosphate dehydrogenase from rat (Rattus norvegicus) erythrocyte. Drug Chem Toxicol.2020:43(2),219-223.
29. [29].Temel, Yusuf, and Umit M. Kocyigit. "Purification of glucose‐6‐phosphate dehydrogenase from rat (Rattus norvegicus) erythrocytes and inhibition effects of some metal ions on enzyme activity." Journal of biochemical and molecular toxicology.2017: 31(9),e21927.
30. [30].Krebs HA. and Eggleston LV. The regulation of the pentosephosphate cycle in rat liver. In: G. Weber, ed. Advances in Enzyme Regulation. Oxford: Pergamon Press Ltd.1978: 12, 421–433.
31. [31].Nelson DL and Cox MM. Lehninger Principles of Biochemistry. 4th ed. New York: WH. Freeman & Co, ABD.2005: 558–561.
32. [32].Stanton RC. Glucose-6-phosphate dehydrogenase, NADPH, and cell survival. IUBMB Life.2012: 64 (5), 362–369.
33. [33].Temel Y, Bozkuş T, Karagözoğlu Y, Çiftci M. "Glutatyon Redüktaz (GR) Enziminin Japon Bıldırcın (Coturnix coturnix japanica) Eritrositlerinden Saflaştırılması ve Karakterizasyonu." Iğdır Univ. J. Inst. Sci. & Tech.2017: 7(3),143-150.
34. [34].Ballatori N, Krance SM, Notenboom S, Shi S, Tieu K, Hammond CL. Glutathione dysregulation and the etiology and progression of human diseases. Biol. Chem.2009:390, 191–214,
35. [35].Couto N, Wood J, and Jill B. "The role of glutathione reductase and related enzymes on cellular redox homoeostasis network." Free radical biology and medicine.2016: 95, 27-42.
36. [36].Işık M, Demir Y, Kırıcı M, Demir R, Şimşek F, Beydemir Ş. Changes in the anti-oxidant system in adult epilepsy patients receiving anti-epileptic drugs. Archives of physiology and biochemistry.2015: 121(3), 97-102.
37. [37].Temel Y, Koçyigit UM, Taysı MŞ, Gökalp F, Gürdere MB, Budak Y, Çiftci M. Purification of glutathione S-transferase enzyme from quail liver tissue and inhibition effects of (3aR, 4S, 7R, 7aS)-2-(4-((E)-3-(aryl) acryloyl) phenyl)-3a, 4, 7, 7a-tetrahydro-1H-4, 7-methanoisoindole-1, 3 (2H)-dione derivatives on the enzyme activity. Journal of biochemical and molecular toxicology.2018: 32(3), e22034.
38. [38].Emek H, Dönmez DB, Bayçu C, Musmul A. Erişkin erkek sıçanlarda karbon tetraklorür ile oluşturulan testis hasarı üzerine kuersetinin etkisi/Effect of quercetin on CCl₄ induced testicular injury in adult male rats. Osmangazi Tıp Dergisi.2014: 39(1),65-73.
39. [39].Elbe H, Eşrefoğlu M, Taslidere A, Taşlıdere E, Ateş B. Sıçanlarda karbon tetraklorür ile indüklenen kardiyak hasar üzerine melatonin ve quercetin'in tedavi edici etkileri. İnönü Üniversitesi Turgut Özal Tıp Merkezi Dergisi.2016: 23(2), 171-176.
40. [40].Ciftci R, Yuce A. Karaciğer fibrozisli ratlarda kuersetinin homosistein düzeyi ve koroner damar hasarı üzerine etkisi. FÜ Sağ Bil Vet Derg.2013: 27(3), 159-67.
41. [41].Cui Y, Han Y, Yang X, Sun Y, Zhao Y. Protective effects of quercetin and quercetin-5', 8-disulfonate against carbon tetrachloride-caused oxidative liver injury in mice. Molecules.2014: 19(1), 291-305.
42. [42].Pavanato A, Tuñón MJ, Sánchez-Campos S, Marroni CA, Llesuy S, González-Gallego J, Marroni N. Effects of quercetin on liver damage in rats with carbon tetrachloride-induced cirrhosis. Digestive diseases and sciences.2013: 48(4), 824-829.
43. [43].Krishnappa P, Venkatarangaiah K, Rajanna S, Kumar S, Kashi Prakash Gupta R. Antioxidant and prophylactic effects of Delonix elata L., stem bark extracts, and flavonoid isolated quercetin against carbon tetrachloride-induced hepatotoxicity in rats. BioMed Research International.2014:1155/507851. [44].Akkoyun HT. Karbontetraklorür’ün Neden Olduğu Sıçan Böbrek Dokusu Hasarlarına Karşı Kuersetin’in Koruyucu Etkisi. Journal of the Institute of Science and Technology.2019: 9(2), 708-716.