A NOVEL AND SEMIAUTOMATED ASSAY FOR THIOL OXIDATIVE STRESS INDEX: TOSI

Abstract

Objective: The aim of the study is to develop a new and automated method determining irreversible and reversible thiol oxidation products which are formed by the exposure of the plasma to oxidation and to measure the resistance of thiol groups to oxidation.

Material and Methods: The method is based on thiol-disulphide homeostasis measurement of plasma thiol groups before and after exposure to oxidative stress. According to the measurements, the levels of reversible thiol oxidation products (RTOP), irreversible thiol oxidation products (ITOP), and thiol oxidative stress index (TOSI=ITOP/RTOP) were determined.

Results: Plasma levels of ITOP were 24.02±12.1 μmol/L, levels of RTOP were 26.68±14.96 μmol/L and TOSI was 0.67±0.60 in the healthy control group. The level of ITOP of patients with lung and colon cancer were found to be significantly higher than the control group, while the level of RTOP was found to be significantly lower. When the parameter of TOSI which was used to determine the direction of the balance was examined, it was observed that the formation of ITOP was more dominant in both colon and lung cancer patients when compared to the control group.

Conclusion: Practical, inexpensive and semi-automatic spectrophotometric RTOP, ITOP and TOSI tests, developed by us for the first time, can be used to investigate many diseases associated with oxidative stress.

Keywords

• Irreversible thiol oxidation products
• Reversible thiol oxidation products
• Thiol

Tiyol Oksidatif Stres İndeksi için Yeni ve Yarı-Otomatik Bir Test: TOSİ

Abstract

Amaç: Çalışmanın amacı, plazmanın oksidasyona maruz kalmasıyla oluşan geri dönüşümsüz ve geri dönüşümlü tiyol oksidasyon ürünlerini belirleyen yeni ve otomatik bir yöntem geliştirmek ve tiyol gruplarının oksidasyona direncini ölçmektir.

Gereç ve Yöntemler: Yöntem, oksidatif strese maruz kalmadan önce ve sonra plazma tiyol gruplarının tiyol-disülfid homeostaz ölçümüne dayanmaktadır. Ölçümlere göre, geri dönüşümlü tiyol oksidasyon ürünleri (RTOP), geri dönüşümsüz tiyol oksidasyon ürünleri (ITOP) ve tiyol oksidatif stres indeksi (TOSİ=ITOP/RTOP) seviyeleri belirlenmiştir.

Bulgular: Sağlıklı kontrol grubunda ITOP plazma düzeyleri 24.02±12.1 µmol/L, RTOP düzeyleri 26.68±14.96 µmol/L ve TOSI düzeyleri 0.67±0.60 olarak bulunmuştur. Akciğer ve kolon kanseri olan hastaların ITOP düzeyleri kontrol grubuna göre anlamlı olarak yüksek iken RTOP düzeyi anlamlı olarak düşük bulunmuştur. Dengenin yönünü belirlemek için kullanılan TOSI parametresi incelendiğinde, ITOP oluşumunun hem kolon hem de akciğer kanserinde kontrol grubuna göre daha baskın olduğu görülmüştür.

Sonuç: İlk defa geliştirdiğimiz pratik, ucuz ve yarı otomatik spektrofotometrik RTOP, ITOP ve TOSI testleri; oksidatif stres ile ilişkili birçok hastalıkta araştırılabilir.

Keywords

• Geri dönüşümlü tiyol oksidasyon ürünleri
• Geri dönüşümsüz tiyol oksidasyon ürünleri
• Tiyol

References

1. 1. Turell L, Radi R, Alvarez B. The thiol pool in human plasma: the central contribution of albumin to redox processes. Free Radic Biol Med. 2013;65:244-53.
2. 2. Poole LB. The basics of thiols and cysteines in redox biology and chemistry. Free Radic Biol Med. 2015;80:148-57.
3. 3. Winther JR, Thorpe C. Quantification of thiols and disulfides. Biochim Biophys Acta. 2014;1840(2):838-46.
4. 4. Hamann MJ. Irreversible oxidation of protein cysteine residues. Retrospective Theses and Dissertations. 2002:1-144.
5. 5. Nemmiche S. Oxidative Signaling Response to Cadmium Exposure. Toxicol Sci. 2017;156(1):4-10.
6. 6. Ates I, Kaplan M, Yuksel M, Mese D, Alisik M, Erel O et al. Determination of thiol/disulphide homeostasis in type 1 diabetes mellitus and the factors associated with thiol oxidation. Endocrine. 2016;51(1):47-51.
7. 7. Kundi H, Ates I, Kiziltunc E, Cetin M, Cicekcioglu H, Neselioglu S et al. A novel oxidative stress marker in acute myocardial infarction; thiol/disulphide homeostasis. Am J Emerg Med. 2015;33(11):1567-71.
8. 8. Dirican N, Dirican A, Sen O, Aynali A, Atalay S, Bircan HA et al. Thiol/disulfide homeostasis: A prognostic biomarker for patients with advanced non-small cell lung cancer? Redox Rep. 2016;21(5):197-203.

9. 9. Dogru A, Balkarli A, Cetin GY, Neselioglu S, Erel O, Tunc SE et al. Thiol/disulfide homeostasis in patients with ankylosing spondylitis. Bosn J Basic Med Sci. 2016;16(3):187-92.
10. 10. Wlodek PJ, Smolenski OB, Chwatko G, Iciek MB, Milkowski A, Bald E et al. Disruption of thiol homeostasis in plasma of terminal renal failure patients. Clin Chim Acta. 2006;366(1-2):137-45.
11. 11. Sbrana E, Paladini A, Bramanti E, Spinetti MC, Raspi G. Quantitation of reduced glutathione and cysteine in human immunodeficiency virus-infected patients. Electrophoresis. 2004;25(10-11):1522-9.
12. 12. Vural G, Gumusyayla S, Bektas H, Deniz O, Alisik M, Erel O. Impairment of dynamic thiol-disulphide homeostasis in patients with idiopathic Parkinson's disease and its relationship with clinical stage of disease. Clin Neurol Neurosurg. 2017;153:50-5.
13. 13. Gumusyayla S, Vural G, Bektas H, Deniz O, Neselioglu S, Erel O. A novel oxidative stress marker in patients with Alzheimer's disease: dynamic thiol-disulphide homeostasis. Acta Neuropsychiatr. 2016;28(6):315-20.
14. 14. Calabrese V, Scapagnini G, Ravagna A, Bella R, Butterfield DA, Calvani M et al. Disruption of thiol homeostasis and nitrosative stress in the cerebrospinal fluid of patients with active multiple sclerosis: evidence for a protective role of acetylcarnitine. Neurochem Res. 2003;28(9):1321-8.
15. 15. Asil M, Dertli R, Biyik M, Yolacan R, Erel O, Neselioglu S et al. Dynamic thiol-disulfide homeostasis is disturbed in patients with non-alcoholic fatty liver disease. Journal of Laboratory Medicine. 2018;42(1-2):31-8.
16. 16. Turell L, Carballal S, Botti H, Radi R, Alvarez B. Oxidation of the albumin thiol to sulfenic acid and its implications in the intravascular compartment. Braz J Medical Biol Res. 2009;42(4):305-11.
17. 17. Ellman G, Lysko H. A precise method for the determination of whole blood and plasma sulfhydryl groups. Anal Biochem. 1979;93(1):98-102.
18. 18. Erel O, Neselioglu S. A novel and automated assay for thiol/disulphide homeostasis. Clin Biochem. 2014;47(18):326-32.
19. 19. Glowacki R, Bald E. Fully automated method for simultaneous determination of total cysteine, cysteinylglycine, glutathione and homocysteine in plasma by HPLC with UV absorbance detection. J Chromatogr B Analyt Technol Biomed Life Sci. 2009;877(28):3400-4.
20. 20. Chen W, Zhao Y, Seefeldt T, Guan X. Determination of thiols and disulfides via HPLC quantification of 5-thio-2-nitrobenzoic acid. J Pharm Biomed Anal.2008;48(5):1375-80. 21. Carru C, Deiana L, Sotgia S, Pes GM, Zinellu A. Plasma thiols redox status by laser-induced fluorescence capillary electrophoresis. Electrophoresis. 2004;25(6):882-9.
21. 22. Mourad T, Min KL, Steghens JP. Measurement of oxidized glutathione by enzymatic recycling coupled to bioluminescent detection. Anal Biochem. 2000;283(2):146-52.
22. 23. van Bergen LA, Roos G, De Proft F. From thiol to sulfonic acid: modeling the oxidation pathway of protein thiols by hydrogen peroxide. J Phys Chem A. 2014;118(31):6078-84.
23. 24. Hanikoglu F, Hanikoglu A, Kucuksayan E, Alisik M, Gocener AA, Erel O et al. Dynamic thiol/disulphide homeostasis before and after radical prostatectomy in patients with prostate cancer. Free Radic Res. 2016;50(sup1):S79-S84.
24. 25. Demirseren DD, Cicek C, Alisik M, Demirseren ME, Aktas A, Erel O. Dynamic thiol/disulphide homeostasis in patients with basal cell carcinoma. Cutan Ocul Toxicol. 2017;36(3):278-82.
25. 26. Guney T, Kanat IF, Alkan A, Alisik M, Akinci S, Silay K et al. Assessment of serum thiol/disulfide homeostasis in multiple myeloma patients by a new method. Redox Rep. 2017;22(6):246-51.
26. 27. Eroglu S, Haskul I, Aziz V, Yurtcu E, Karatas F, Neselioglu S et al. Dynamic thiol/disulphide homeostasis in patients with Uterine Myoma. Eur J Obstet Gynecol Reprod Biol. 2017;216:24-6.
27. 28. Manda G, Isvoranu G, Comanescu MV, Manea A, Debelec Butuner B, Korkmaz KS. The redox biology network in cancer pathophysiology and therapeutics. Redox Biolog. 2015;5:347-57.
28. 29. Morrison DK, Davis RJ. Regulation of MAP kinase signaling modules by scaffold proteins in mammals. Annu Rev Cell Dev Biol. 2003;19:91-118.
29. 30. Eijkelenboom A, Burgering BM. FOXOs: signalling integrators for homeostasis maintenance. Nat Rev Mol Cell Biol. 2013;14(2):83-97.