In VitroEffects of Some Drugs on Glucose-6-Phosphate Dehydrogenase Enzyme Purified from Sheep Spleen Tissue

Year 2022, , 29 - 35, 25.03.2022

Çiğdem Çoban , Mehmet Çiftci

https://doi.org/10.46810/tdfd.929840

Cited By: 1

Abstract

In this study, in vitroeffects of some drugs on glucose-6-phosphate dehydrogenase enzyme (G6PD; E.C. 1.1.1.49) purified from sheep spleen tissue were investigated. In the first step of the study, G6PD enzyme was purified from sheep spleen tissue by salting out(ammonium sulfateprecipitation) method and 2', 5' ADP-Sepharose 4B gel affinity chromatography and the purity of the enzyme was checked by SDS-PAGE method. In the second phase of the study, the effects of iveral, lincomycin, gentamicin, amoxicillin, ampicillin, streptomycin sulfate, novamizole, ketogesic, cefuroxime, cefazolin sodium and tylosin drugs on enzyme activity were investigated. Research results showed that iveral, gentamicin, streptomycin sulfate and lincomycin drugs inhibit the G6PD enzyme with IC50values of 0.62 mM, 21.6 mM, 173.2 mM and 231 mM, respectively, amoxicillin, ampicillin, novamizole, ketogenic, cefuroxime sodium, cefazolin sodium and tylosin drugs did not have a significant activation or inhibition effect on the enzyme.

Keywords

Glucose 6-phosphate dehydrogenase, Purification, Drug, Inhibition, Iveral, Gentamicin

Bazı İlaçların Koyun Dalak Dokusundan Saflaştırılan Glukoz-6-Fosfat Dehidrogenaz Enzimi Üzerine İn Vitro Etkileri

Abstract

Bu çalışmada bazı ilaçların, koyun dalak dokusundan saflaştırılan glukoz-6-fosfat dehidrogenaz enzimi (G6PD; E.C. 1.1.1.49) üzerine in vitroetkileri araştırıldı. Çalışmanın ilk aşamasında G6PD enzimi koyun dalak dokusundan salting out(amonyum sülfat çöktürmesi) metodu ve 2', 5' ADP-Sepharose 4B afinite kromatografisi ile saflaştırıldı ve enzimin saflık derecesi SDS-PAGE metodu ile kontrol edildi. Çalışmanın ikinci aşamasında iveral, linkomisin, gentamisin, amoksisilin, ampisilin, streptomisin sülfat, novamizol, ketojezik, sefuroksim, sefazolin sodyum ve tylosin ilaçlarının enzim aktivitesi üzerindeki etkileri araştırıldı. Araştırma sonuçları; iveral, gentamisin, streptomisin sülfat ve linkomisin ilaçlarının G6PD enzimini sırasıyla 0,62 mM, 21,6 mM, 173,2 mM ve 231 mM, IC50değerleri ile inhibe ettiğini, amoksisilin, ampisilin, novamizol, ketojezik, sefuroksim sodyum, sefazolin sodyum ve tylosin ilaçlarının ise enzim üzerinde önemli bir aktivasyon ya da inhibisyon etkisine sahip olmadıklarını gösterdi.

Keywords

glukoz 6-fosfat dehidrogenaz, saflaştırma, ilaç, inhibisyon, iveral, gentamisin

References

• [1]Ciftci M, Bilici D, Kufrevioglu OI. Effects of metamizol and magnesium sulfate on enzyme activities of glucose-6-phosphate dehydrogenase from human erythrocytes in vitro and from rat erythrocytes in vivo. Pharmacol Res. 2001;44(1):7–11.
• [2]Adem S, Ciftci M. Purification and biochemical characterization of glucose 6-phosphate dehydrogenase, 6-phosphogluconate dehydrogenase and glutathione reductase from rat lung and inhibition effects of some antibiotics. J Enzyme Inhib Med Chem. 2016; 31(6):1342-1348.
• [3]Temel Y, Kocyigit UM. Purification of glucose-6-phosphate dehydrogenase from rat (Rattus norvegicus) erythrocytes and inhibition effects of some metal ions on enzyme activity. J Biochem Mol Toxicol. 2017;31(9): e21927.
• [4]Gumustekin K, Ciftci M, Coban A, Altikat S, Aktas O, Gul M, et al. Effects of nicotine and vitamin E on glucose 6-phosphate dehydrogenase activity in some rat tissues in vivo and in vitro. J Enzyme Inhib Med Chem. 2005;20(5):497–502.
• [5]Temel Y, Ayna A, Hamdi Shafeeq I, CiftciM. In vitro effects of some antibiotics on glucose-6-phosphate dehydrogenase from rat (Rattus norvegicus) erythrocyte. Drug Chem Toxicol [Internet]. 2020;43(2):219–23.
• [6]Temel Y, Bayındır S. The Synthesis of Thiosemicarbazone-Based Aza-Ylides as Inhibitors of Rat Erythrocyte Glucose 6-Phosphate Dehydrogenase Enzyme. J Inst Sci Technol. 2019;9(3):1503–12.
• [7]Pljesa-Ercegovac M, Savic-Radojevic A, Matic M, Coric V, Djukic T, Radic T, et al. Glutathione transferases: Potential targets to overcome chemoresistance in solid tumors. Int J Mol Sci. 2018;19(12).
• [8]Lu J, Holmgren A. Free Radical Biology and Medicine The thioredoxin antioxidant system. Free Radic Biol Med [Internet]. 2014;66:75–87. Available from:http://dx.doi.org/10.1016/j.freeradbiomed.2013.07.036
• [9]Taysi MŞ, Temel Y. Glutathione S-transferase: Purification and Characterization from Quail (Coturnix coturnix japonica) Liver and the Impact of Some Metal Ions on Enzyme Activity. Bionanoscience. 2021;11(1):91–8.
• [10]Arnér ESJ, Holmgren A. The thioredoxin system in cancer. Semin Cancer Biol. 2006;16(6):420–6.
• [11]Carvalho CML, Chew E, Hashemy SI, Lu J, Holmgren A. Inhibition of the Human Thioredoxin System A Molecular Mechanısm Of Mercury Toxıcıty. 2008;283(18):11913–23.
• [12]Temel Y, Kufrevioǧlu ÖI, Çiftci M. Investigation of the effects of purification and characterization of turkey (Meleagris gallopavo) liver mitochondrial thioredoxin reductase enzyme and some metal ions on enzyme activity. Turkish J Chem. 2017;41(1).
• [13]Akkemik E, Şentürk M, Özgeriş FB, Taşer P, Çiftci M. In vitro eff ects of some drugs on human erythrocyte glutathione reductase. 2011;41(2):235–41.
• [14]Dilek EB, Ömer İ, Beydemir Ş. mpacts of some antibiotics on human serum paraoxonase 1 activity. 2013;28(February 2012):758–64.
• [15]Zhang D, Liu Y, Luo Z, Chen Y, Xu A, Liang Y, et al. The novel thioredoxin reductase inhibitor A-Z2 triggers intrinsic apoptosis and shows efficacy in the treatment of acute myeloid leukemia. Free Radic Biol Med. 2020;146(November 2019):275–86.
• [16]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.
• [17]Shapiro R, Vallee BL. Interaction of Human Placental Ribonuclease with Placental Ribonuclease Inhibitor. Biochemistry. 1991;30(8):2246–55.
• [18]Kose LP, Gülçin İ, Özdemir H, Atasever A, Alwasel SH, Supuran CT. The effects of some avermectins on bovine carbonic anhydrase enzyme. J Enzyme Inhib Med Chem [Internet]. 2015;6366(February 2017):1–6. Available from: http://www.ncbi.nlm.nih.gov/pubmed/26207514
• [19]Adem S, Ciftci M. Purification of rat kidney glucose 6-phosphate dehydrogenase, 6-phosphogluconate dehydrogenase, and glutathione reductase enzymes using 2',5'-ADP Sepharose 4B affinity in a single chromatography step. Protein Expr Purif. 2012;81(1):1–4.
• [20]Özaslan MS, Balcı N, Demir Y, Gürbüz M, Küfrevioğlu Öİ. Inhibition effects of some antidepressant drugs on pentose phosphate pathway enzymes. Environ Toxicol Pharmacol. 2019;72(March).
• [21]Temel Y, Ciftci M, Akkoyun HT, Akkoyun M. Effect of astaxanthin and aluminum chloride on erythrocyte G6PD and 6PGD enzyme activities in vivo and on erythrocyte G6PD in vitro in rats. 2017;31(10):e21954.
• [22]Temel Y, Taysi MŞ. The Effect of Mercury Chloride and Boric Acid on Rat Erythrocyte Enzymes. Biol Trace Elem Res. 2019;191(1)172-187.
• [23]Altikat S, Ciftci M, Buyukokuroglu ME. In vitro effectsof some anesthetic drugs on enzymatic activity of human red blood cell glucose 6-phosphate dehydrogenase\r758. Pol J Pharmacol [Internet]. 2002;54(1):67–71.
• [24]Temel Y, Bozkuş T, Karagözoğlu Y, Çİftcİ M. Glutatyon Redüktaz ( GR ) Enziminin Japon Bıldırcın ( Coturnix coturnix japanica ) Eritrositlerinden Saflaştırılması ve Karakterizasyonu Purification and Characterization of Glutathion Reductase Enzyme From Japanese Quail ( Coturnix coturnix japanica ) Er. 2017;7(3):143–50.
• [25]Perosa F, Carbone R, Ferrone S, Dammacco F. Purification of human immunoglobulins by sequential precipitation with caprylic acid and ammonium sulphate. J Immunol Methods. 1990;128(1):9–16.
• [26]Mannervik B, Jacobsson K, Boggaram V. Department of Biochemistry, Arrhenius Laboratory, University of Stockholm, Stockholm, Sweden. 1976;66(2):221–4.
• [27]Buyukokuroglu ME. the Effects of Ethanol on Glucose 6-Phosphate Dehydrogenase Enzyme Activity From Human Erythrocytes in Vitro and Rat Erythrocytes in Vivo. Alcohol Alcohol. 2002;37(4):327–9.
• [28]Bradford MM. Interactive effects of drought stress and chitosan application on physiological characteristics and essential oil yield of Thymus daenensis Celak. Crop J. 2017;5(5):407–15.
• [29]Laemmli UK. 1970 Nature Publishing Group. Nat Publ Gr. 1970;228:1979.
• [30]Lineweaver H, Burk D. The Determination of Enzyme Dissociation Constants. J Am Chem Soc. 1934;56(3):658–66.
• [31]Jiang P, Du W, Wu M. Regulation of the pentose phosphate pathway in cancer. Protein Cell. 2014;5(8):1–11.
• [32]Patra KC, Hay N. The pentose phosphate pathway and cancer. Trends Biochem Sci. 2014;39(8):347–54.
• [33]Luo Z, Du D, Liu Y, Lu T, Liu L, Jiang H, et al. Discovery and characterization of a novel glucose-6-phosphate dehydrogenase (G6PD) inhibitor via high-throughput screening. Bioorg Med Chem Lett. 2021;40(February):127905.
• [34]Goa Karen L, Donna McTavish, Stephen PC. Ivermectin: A review of its antifilarial activity, pharmacokinetic properties and clinical efficacy in onchocerciasis. Drugs. 1991;42(4):640–58.
• [35]Sader HS, Farrell DJ, Flamm RK, Jones RN. Antimicrobial susceptibility of Gram-negative organisms isolated from patients hospitalized in intensive care units in United States and European hospitals (2009-2011). Diagn Microbiol Infect Dis. 2014;78(4):443–8.
• [36]Kumar M, Kakkar V, Mishra AK, Chuttani K, Kaur IP. Intranasal delivery of streptomycin sulfate (STRS) loaded solid lipid nanoparticles to brain and blood. Int J Pharm. 2014;461(1–2):223–33.
• [37]Leclercq R, Courvalin P. Erratum: Bacterial resistance to macrolide, lincosamide, and streptogramin antibiotics by target modification (Antimicrobial Agents and Chemotherapy 35 (1268)). Antimicrob Agents Chemother. 1991;35(10):2165.
• [38]Poole K. Resistance to β-lactam antibiotics. Cell Mol Life Sci. 2004;61(17):2200–23.
• [39]El-Shaboury SR, Saleh GA, Mohamed FA, Rageh AH. Analysis of cephalosporin antibiotics. J Pharm Biomed Anal. 2007;45(1):1–19.
• [40]Gökçimen A, Özgüner M, Bayram D, Ural M, Sulak O. Metamizol sodyumun sıçan karaciğer, böbrek ve akciğer dokuları üzerine etkisi. SDÜ Tıp Fakültesi Derg. 2009;13(1):27–31.
• [41]Alkimin GD, Soares AMVM, Barata C, Nunes B. Evaluation of ketoprofen toxicityin two freshwater species: Effects on biochemical, physiological and population endpoints. Environ Pollut. 2020;265:114993.
• [42]Hendrickson OD, Zvereva EA, Zherdev A V., Godjevargova T, Xu C, Dzantiev BB. Development of a double immunochromatographic test system for simultaneous determination of lincomycin and tylosin antibiotics in foodstuffs. Food Chem. 2020;318(February):126510.
• [43]Adem S, Ciftci M. Purification of rat kidney glucose 6-phosphate dehydrogenase, 6-phosphogluconate dehydrogenase, and glutathione reductase enzymes using 2???,5???-ADP Sepharose 4B affinity in a single chromatography step. Protein Expr Purif.2012;81(1):1–4.
• [44]Aksoy M, Ozaslan MS, Kufrevioglu OI. 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. 2016;31(4):546–50.
• [45]Ceylan M, Temel Y, Kocyigit UM, Alwasel SH, Gülçin I, Gürbüzlü B. Synthesis , carbonic anhydrase I and II isoenzymes inhibition properties , and antibacterial activities of novel tetralone-based 1 , 4-benzothiazepine derivatives. 2017;31(4):e21872.
• [46]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. J Biochem Mol Toxicol. 2018;32(9):1–9.
• [47]Bayindir S, Ayna A, Temel Y, Çiftci 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 J Chem. 2018;42(2):332-345.
• [48]Beydemir S, Ciftçi M, Küfrevioğlu OI. Purification and characterization of glucose 6-phosphate dehydrogenase from sheep erythrocytes and inhibitory effects of some antibiotics on enzyme activity. J Enzyme Inhib Med Chem. 2002;17(4):271–7.
• [49]Caglayan C. The toxicological effects of some avermectins on goat liver carbonic anhydrase enzyme. 2018;32(1):e22010.