Mehmet ÇİFTÇİ, Murat ŞENTÜRK, Fatma Betül ÖZGERİŞ, Ebru AKKEMİK, Pınar TAŞER

In vitro eff ects of some drugs on human erythrocyte glutathione reductase

Amaç: Bu çalışmada dekarbazin, tiyokolsikosid, metotreksat, olanzapin, sodium pantoprozol ve 5-fl orourasil ilaçlarının insan eritrositlerinden safl aştırılan glutatyon redüktaz enzimi (GR) üzerine in vitro etkileri araştırıldı. Yöntem ve gereç: Bu amaçla 4 EÜ/mg protein spesifi k aktivitesine sahip olan enzim sırasıyla hemolizatın hazırlanması, amonyum sülfat çöktürmesi, Sephadex G-200 jel fi ltrasyon kromatografi si ve 2’,5’-ADP Sepharose 4B afi nite kromatografi si ile % 44,44 verimle 3333 kat safl aştırıldı. Bu dört basamak sonucu elde edilen saf enzimin safl ık kontrolü sodyum dodesilsülfat poliakrilamid jel elektroforezzi (SDS-PAGE) ile yapıldı. Tüm kinetik çalışmalarda safl aştırılmış enzim kullanıldı. Bulgular: Enzim üzerinde inhibisyon etkisi gösteren dekarbazin, tiyokolsikosid, metotreksat, olanzapin, sodium pantoprozol ve 5-fl orourasil ilaçları için sırasıyla IC50 değerleri ve Ki sabitleri 0,014 ve 0,0121 ± 0,0026; 0,054 ve 0,0135 ± 0,0024; 0,31 ve 0,1213 ± 0,040; 0,35 mM ve 0,3173 ± 0,0981; 0,94 ve 0,5413 ± 0,0728; 98,57 ve 69,611 ± 9,690; olarak hesaplandı. Ayrıca inhibisyon çeşidi çalışmaları sonucu tüm ilaçların enzimi yarışmasız olarak inhibe ettikleri tespit edildi. Sonuç: Kullanılan tüm ilaçların enzim üzerinde inhibisyon etkisi tespit edildi.

Bazı ilaçların insan eritrosit glutatyon redüktaz enzimi üzerine etkileri

Aim: Th e eff ects of dacarbazine, thiocolchicoside, methotrexate, olanzapine, pantoprazole sodium, and 5-fl uorouracil on the enzyme activity of glutathione reductase (GR) were studied using human erythrocyte GR enzymes in in vitro studies, respectively. Materials and methods: Th e enzyme was purifi ed 3333-fold from human erythrocytes in a yield of 44.44% with 4.0 U/mg. Th e purifi cation procedure involved the preparation of hemolysate, ammonium sulfate precipitation, 2’, 5’-ADP Sepharose 4B affi nity chromatography, and Sephadex G-200 gel fi ltration chromatography. SDS-PAGE showed a single band for enzyme. Purifi ed enzyme was used in the in vitro studies. Results: In the in vitro studies, IC50 values and Ki constants were 0.014 mM and 0.0121 ± 0.0026 mM for dacarbazine, and 0.054 mM and 0.0135 ± 0.0024 mM for thiocolchicoside, 0.31 mM and 0.1213 ± 0.040 mM for methotrexate, 0.35 mM and 0.3173 ± 0.0981 mM for olanzapine, 0.94 mM and 0.5413 ± 0.0728 mM for pantoprazole sodium, and 98.57 mM and 69.611 ± 9.690 mM for 5-fl uorouracil, showing the inhibition eff ects on the purifi ed enzyme. Inhibition types were non-competitive for dacarbazine, methotrexate, olanzapine, and 5-fl uorouracil and competitive for thiocolchicoside and pantoprazole sodium. Conclusion: All the drugs indicated the inhibitory eff ects on the enzyme.

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