İnsan eritrosit glukoz 6-fosfat dehidrogenaz enzim aktivitesi üzerine bazı antibiyotiklerin in vitro etkileri

Amaç : İnsan eritrositlerinden elde edilen glukoz 6-fosfat dehidrogenaz enzimi üzerine bazı antibiyotiklerin inhibisyon etkilerini araştırmak. Yöntem : Bu amaçla önce eritrosit glukoz 6-fosfat dehidrogenaz enzimi, 2',5'-ADP Sepharose 4B jelinin kullanıldığı afinite kromatografisi vasıtasıyla % 28 verimle 13.654 kat saflaştırıldı. Saflaştırma esnasında sıcaklık +2 o C'de kontrol altında tutuldu. Enzim aktivitesi Beutler metoduna göre 340 nm'de spektrofotometrik olarak belirlendi ve bütün kinetik çalışmalarda bu metod uygulandı. İlaç olarak seftriakson disodyum trisesquihidrat, penicillin G potasyum, amoksisilin sodyum kullanıldı. Bulgular : Bu ilaçların glukoz 6-fosfat dehidrogenaz enzimi üzerinde inhibisyon etkileri olduğu tesbit edildi. İnhibisyon etkisi gösteren bu ilaçlar için % aktivite-[I] grafikleri çizilerek I 50 değerleri bulundu. Sonuç : Glukoz 6-fosfat dehidrogenaz eksikliği olmayan sağlıklı kişilerde bu ilaçların kullanılması fazla sakıncalı olmamakla birlikte, glukoz 6-fosfat dehidrogenaz enzim eksikliği olan kişilerde, özellikle seftriakson disodyum trisesquihidrat kullanılmasının ciddi sağlık problemlerine yol açabilir.

In vitro effects of some antibiotics on enzymatic activity of human erythrocytes glucose 6-phosphate dehydrogenase

Objective : Inhibitory effects of some antibiotics on glucose 6-phosphate dehydrogenase enzyme isolated from human erythrocytes was researched. Methods : Erythrocyte glucose 6-phosphate dehydrogenase was purified 13.654 times with 28% yield by affinity chromatography in which 2',5'-ADP Sepharose 4B gel was used. During purification process, temperature was stabilized at +2 o C.Enzyme activity was determined by using spectrophotometer at 340 nm according to Beutler method. Ceftriaxone disodium trisesquihydrate, penicillin G potassium, amoxicillin sodium were used as antibiotics. Results : It was understood that all these drugs have inhibitory effects on glucose 6-phosphate dehydrogenase enzyme. I 50 values were found for these antibiotics by drawing % activity-[I] graphics. Conclusion : Although it does not cause any serious problem to use these drugs people who do not have G6PD deficiency, especially ceftriaxone disodium trisesquihydrate may cause serious health problems in G6PD deficient people.

___

  • 1. Lehninger AL, Nelson DL, Cox MM. Principles of Biochemistry. New York: Worth Publishers, 1993.
  • 2. Keha EE, Küfrevioğlu Öİ. Biyokimya. Erzurum: Şafak Yayınevi; 1997.
  • 3. Reuter R, Naumann M, Metz P, Kopperschlager G. Purification and characterization of glucose 6-phosphate dehydrogenase from Pseudomonas W6. Biomed Biochem Acta 1990; 7: 539-46.
  • 4. Yüreğir TG, Aksoy K, Dikmen N, Ünlükurt İ. Glukoz 6-fosfat dehidrogenaz enziminin insan eritrositlerinden kısmi saflaştırılması ve enzimolojik özelliğinin incelenmesi. İçinde: IX. Ulusal Biyoloji Kongresi bildiri kitabı. 1988. p.125-130.
  • 5. Yoshida A, Huang IY. Structure of human G6PD.London: Academic Press; 1986.
  • 6. Deutsch J. Glucose-6-phosphate dehydrogenase. In: Bergmeyer HU, Bergmeyer J, editors. Methods of enzymatic analysis. 1983. p.3:190-6.
  • 7. Weksler BB, Moore A, Tepler J. Hematology. In: Andreoli TE, Carpenter CCJ, Plum F, Smith LH Jr, editors. Cecil essentials of medicine. 2nd Ed. Philadelphia: WB Saunders; 1990. p.341-63.
  • 8. Aksoy K. Glukoz 6-fosfat dehidrogenaz enziminin yapısı. Adana: Çukurova Üniversitesi Tıp Fakültesi Yayınları; 1997.
  • 9. Yüreğir GT, Aksoy K, Arpaci A, Ünlükurt İ, Tuli A. Studies on red cell glucose-6-phosphate dehydrogenase, evaluation of reference values. Ann Clin Biochem 1994;31:50-5.
  • 10. Berkow R (Editor-in-Chief). The merck manual of diagnosis and therapy. 15th Ed. USA: Merck and Co; 1987.
  • 11. Laurence DR, Bennett PN, Brown MJ. Clinical pharmacology. 8th Ed. Singapore: Churchill Livingstone; 1997.
  • 12. Beutler E. Glucose 6-phosphate dehidrogenase deficiency and other enzyme abnormalities. In: Beutler E, Lichtman MA, Coller BF, Kipps TJ. editors. Williams hematology. 5th Ed. New York: Mc Graw-Hill; 1995. p.564-80.
  • 13. Telefoncu A, Zihnioğlu F. Glukoz 6-fosfat dehidrogenaz aktivitesine primaquin'in etkisi. Tr J Med Sci 1990;14:57-63.
  • 14. Kayaalp SO. Rasyonal tedavi yönünden tıbbi farmakoloji. Ankara: Hacettepe-Taş yayıncılık; 1998.
  • 15. Webster LT Jr. Drugs used in the chemothrapy of protozoal infections (malaria). In: Goodman Gilman A, Rall WT, Nies AS, Taylor P, editors. Goodman-Gilman's The pharmacological basis of therapeutics. 8th Ed. USA: Pergamon press; 1990. p.978-98.
  • 16. Çiftçi M, Küfrevioğlu Öİ, Gündoğdu M, Özmen İ. Effects of some antibiotics on enzyme activity of glucose-6-phosphate dehydrogenase from human erythrocytes. Pharmacol Res 2000;41:109-13.
  • 17. Ninfali P, Orsenigo T, Barociani L, Rapa S. Rapid purification of glucose 6-phosphate dehydrogenase from mammal's erythrocytes. Preparative Biochemistry 1990;20:297-309.
  • 18. Delgado C, Tejedor MJ Luque J. Partial purification of glucose 6-phosphate dehydrogenase and phosphofructokinase from rat erythrocyte haemolysate by partitioning in aqueous two-phase systems. J Chromatograpy 1990;498: 159-68.
  • 19. Shreve DS, Levy HR. On the molecular weight of human glucose 6-phosphate dehydrogenase. Biochem Biophys Res Comm 1997;78:1369-75.
  • 20. Morelli A, Benatti U, Gaetani GF, De Flora A. Biochemical mechanisms of glucose 6-phosphate dehydrogenase deficiency. Proc Natl Acad Sci 1978;75:4-9.
  • 21. Beutler E. Red cell metabolism manual of biochemical methods. London: Academic Press; 1971.
  • 22. Warnock DG. Diuretic agent. In: Katzung BG, editor. Basic and clinical pharmacology. 4th Ed. USA: Appletion and Lange; 1989. p.183-97.
  • 23. Gupta BL, Nehal M, Baquer NZ. Effect of experimental diabetes on the activities of hexokinase, glucose-6-phosphate dehydrogenase and catecolamines in rat erythrocytes of different ages. Indian J Exp Biol 1997;35:792-5.
  • 24. Bekyarova G, Yankova T, Kozarev I. Combined application of alpha-tocopherol and FC-43 perfluorocarbon emulsion suppresses early postburn lipid peroxidation and improves deformability of erythrocytes. Acta Chir Plast 1998;40:17-21.
  • 25. Firguet B, Stadtman ER, Szweda LJ. Modification of glucose-6-phosphate dehydrogenase by 4-hydroxy-2-nonenal. J Biol Chem 1994;269:26.
  • 26. Bessis M, Weed R, Leblond P. Red cell shape, Physiology, Pathology, Ultrastructure. New York: Spriger Verlag; 1973.
  • 27. Beutler E. Red cell enzyme defects as nondisease and as disease. Blood 1979;54:1.
  • 28. Beutler E. Glucose-6-phosphate dehydrogenase deficiency. Blood 1994;84:3613-36.
  • 29. Barza M, Miao PVW. Antimicrobial spectrum. Pharmacology and therapeutic use of antibiotics. Am J Hosp Pharm 1977;34:621.
  • 30. Halliwell B. Free radicals, antioksidants, and human disease: curiosity, cause, or consequence? The Lancet 1994;344:721-24.