Urinary biomarkers of oxidative stress in amyotrophic lateral sclerosis – a pilot study
Authors:
P. Malá 1,2; O. Vyšata 1,2; N. Váňová 3
Authors‘ workplace:
Neurologická klinika LF UK, Hradec Králové
1; Neurologická klinika FN Hradec Králové
2; Katedra farmaceutické chemie a farmaceutické analýzy, Farmaceutická fakulta UK, Hradec Králové
3
Published in:
Cesk Slov Neurol N 2026; 89(4): 257-262
Category:
Original Paper
doi:
https://doi.org/10.48095/cccsnn2026257
Overview
Aim: The aim of this pilot study was to determine whether urinary biomarkers of oxidative damage in patients with amyotrophic lateral sclerosis (ALS) correlate with disease severity, disease duration, and survival prospectively measured from the time of urinary biomarker assessment. Urine was chosen as a noninvasive medium suitable for repeated sampling. Patients and methods: A total of 34 patients with ALS diagnosed according to the Gold Coast criteria were included. Urinary concentrations of 3 nitrotyrosine, 8 hydroxy 2’ deoxyguanosine, malondialdehyde, reduced glutathione, free thiol groups and their disulfide forms, representing protein, DNA and lipid damage and alterations in antioxidant systems, were measured using liquid chromatography coupled with tandem mass spectrometry, high performance liquid chromatography and spectrophotometry. Associations between biomarkers and clinical parameters were assessed using non parametric correlations and quadratic regression. Results: Disease duration was significantly associated with higher 8 hydroxy 2’ deoxyguanosine levels (Spearman r = 0.34; P = 0.049; Kendall t = 0.25; P = 0.050) and lower reduced glutathione levels, with a marked inverse correlation between reduced glutathione and disease duration (Spearman r = −0.59; P = 0.00016; Kendall t = −0.41; P = 0.00057). No significant association was found between any of the evaluated biomarkers and patients’ functional status or survival time. Conclusion: Urinary levels of 8 hydroxy 2’ deoxyguanosine and reduced glutathione show an association with ALS duration and suggest the potential use of urinary biomarkers for non invasive assessment of this disease, although no relationship with functional status or survival time was demonstrated. These findings are consistent with the hypothesis that oxidative stress contributes to ALS pathogenesis, but their clinical relevance needs to be confirmed in larger longitudinal studies.
Keywords:
Antioxidants – Urine – Glutathione – Amyotrophic lateral sclerosis – Thiols – oxidative stress – biomarkers – DNA damage – 8 hydroxy 2’ deoxyguanosine
Sources
1. Hardiman O, Al-Chalabi A, Chio A et al. Amyotrophic lateral sclerosis. Nat Rev Dis Primers 2017; 3 : 17071. doi: 10.1038/nrdp.2017.71.
2. Kiernan MC, Vucic S, Cheah BC et al. Amyotrophic lateral sclerosis. Lancet 2011; 377 (9769): 942–955. doi: 10.1016/S0140-6736 (10) 61156-7.
3. Singh A, Kukreti R, Saso L et al. Oxidative stress: a key modulator in neurodegenerative diseases. Molecules 2019; 24 (8): 1583. doi: 10.3390/molecules24081583.
4. Chang Y, Kong Q, Shan X et al. Messenger RNA oxidation occurs early in disease pathogenesis and promotes motor neuron degeneration in ALS. PLoS One 2008; 3 (7): e2849. doi: 10.1371/journal.pone.0002849.
5. Warita H, Hayashi T, Murakami T et al. Oxidative damage to mitochondrial DNA in spinal motoneurons of transgenic ALS mice. Mol Brain Res 2001; 89 (1): 147–152. doi: 10.1016/S0169-328X (01) 00029-8.
6. Sau D, De Biasi S, Vitellaro-Zuccarello L et al. Mutation of SOD1 in ALS: a gain of a loss of function. Hum Mol Genet 2007; 16 (13): 1604–1618. doi: 10.1093/hmg/ddm110.
7. Zou ZY, Zhou ZR, Che CH et al. Genetic epidemiology of amyotrophic lateral sclerosis: a systematic review and meta-analysis. J Neurol Neurosurg Psychiatry 2017; 88 (7): 540–549. doi: 10.1136/jnnp-2016-315018.
8. Mitsumoto H, Garofalo DC, Santella RM et al. Plasma creatinine and oxidative stress biomarkers in amyotrophic lateral sclerosis. Amyotroph Lateral Scler Frontotemporal Degener 2020; 21 (3–4): 263–272. doi: 10.1080/21678421.2020.1746810.
9. Wang Z, Bai Z, Qin X et al. Aberrations in oxidative stress markers in amyotrophic lateral sclerosis: a systematic review and meta-analysis. Oxid Med Cell Longev 2019; 2019 : 1712323. doi: 10.1155/2019/1712323.
10. Tohgi H, Abe T, Yamazaki K et al. Remarkable increase in cerebrospinal fluid 3-nitrotyrosine in patients with sporadic amyotrophic lateral sclerosis. Ann Neurol 1999; 46 (1): 129–131. doi: 10.1002/1531-8249 (199907) 46 : 1<129:: AID-ANA21>3.0.CO; 2-W.
11. Murata T, Ohtsuka C, Terayama Y. Increased mitochondrial oxidative damage in patients with sporadic amyotrophic lateral sclerosis. J Neurol Sci 2008; 267 (1–2): 66–69. doi: 10.1016/j.jns.2007.09.038.
12. Bogdanov M, Brown RH, Matson W et al. Increased oxidative damage to DNA in ALS patients. Free Radic Biol Med 2000; 29 (7): 652–658. doi: 10.1016/S0891-5849 (00) 00349-X.
13. Baillet A, Chanteperdrix V, Trocmé C et al. The role of oxidative stress in amyotrophic lateral sclerosis and Parkinson’s disease. Neurochem Res 2010; 35 (10): 1530–1537. doi: 10.1007/s11064-010-0212-5.
14. Bandookwala M, Sengupta P. 3-Nitrotyrosine: a versatile oxidative stress biomarker for major neurodegenerative diseases. Int J Neurosci 2020; 130 (10): 1047–1062. doi: 10.1080/00207454.2020.1713776.
15. Vucic S, Ferguson TA, Cummings C et al. Gold Coast diagnostic criteria: implications for ALS diagnosis and clinical trial enrollment. Muscle Nerve 2021; 64 (5): 532–537. doi: 10.1002/mus.27392.
16. Váňová N, Múčková L, Kalíšková T et al. In vitro evaluation of oxidative stress induced by oxime reactivators of acetylcholinesterase in HepG2 cells. Chem Res Toxicol 2023; 36 (12): 1912–1920. doi: 10.1021/acs.chemrestox.3c00203.
17. Vanova N, Muckova L, Schmidt M et al. Simultaneous determination of malondialdehyde and 3-nitrotyrosine in cultured human hepatoma cells by liquid chromatography-mass spectrometry. Biomed Chromatogr 2018; 32 (12): e4349. doi: 10.1002/bmc.4349.
18. Radi R. Oxygen radicals, nitric oxide, and peroxynitrite: redox pathways in molecular medicine. Proc Natl Acad Sci U S A 2018; 115 (23): 5839–5848. doi: 10.1073/pnas.1804932115.
19. Schwemmer M, Fink B, Köckerbauer R et al. How urine analysis reflects oxidative stress – nitrotyrosine as a potential marker. Clin Chim Acta 2000; 297 (1–2): 207–216. doi: 10.1016/S0009-8981 (00) 00247-3.
20. Valavanidis A, Vlachogianni T, Fiotakis C. 8-Hydroxy-2’-deoxyguanosine (8-OHdG): a critical biomarker of oxidative stress and carcinogenesis. J Environ Sci Health C Environ Carcinog Ecotoxicol Rev 2009; 27 (2): 120–139. doi: 10.1080/10590500902885684.
21. Wu LL, Chiou CC, Chang PY et al. Urinary 8-OHdG: a marker of oxidative stress to DNA and a risk factor for cancer, atherosclerosis and diabetics. Clin Chim Acta 2004; 339 (1–2): 1–9. doi: 10.1016/j.cccn.2003.09.010.
22. Janero DR. Malondialdehyde and thiobarbituric acid-reactivity as diagnostic indices of lipid peroxidation and peroxidative tissue injury. Free Radic Biol Med 1990; 9 (6): 515–540. doi: 10.1016/0891-5849 (90) 90131-2.
23. Del Rio D, Stewart AJ, Pellegrini N. A review of recent studies on malondialdehyde as toxic molecule and biological marker of oxidative stress. Nutr Metab Cardiovasc Dis 2005; 15 (4): 316–328. doi: 10.1016/j.numecd.2005.05.003.
24. Tsikas D. Assessment of lipid peroxidation by measuring malondialdehyde (MDA) and relatives in biological samples: analytical and biological challenges. Anal Biochem 2017; 524 : 13–30. doi: 10.1016/j.ab.2016.10.021.
25. Forman HJ, Zhang H, Rinna A. Glutathione: overview of its protective roles, measurement, and biosynthesis. Mol Aspects Med 2009; 30 (1–2): 1–12. doi: 10.1016/j.mam.2008.08.006.
26. Wu G, Fang YZ, Yang S et al. Glutathione metabolism and its implications for health. J Nutr 2004; 134 (3): 489–492. doi: 10.1093/jn/134.3.489.
27. Meister A, Anderson ME. Glutathione. Annu RevBiochem 1983; 52 : 711–760. doi: 10.1146/annurev.bi.52. 070183.003431.
28. Lash LH. Renal glutathione: dual roles as antioxidant protector and bioactivation promoter. Biochem Pharmacol 2024; 228 : 116181. doi: 10.1016/j.bcp.2024.116181.
29. Ulrich K, Jakob U. The role of thiols in antioxidant systems. Free Radic Biol Med 2019; 140 : 14–27. doi: 10.1016/j.freeradbiomed.2019.05.035.
30. Erel O, Neselioglu S. A novel and automated assay for thiol/disulphide homeostasis. Clin Biochem 2014; 47 (18): 326–332. doi: 10.1016/j.clinbiochem.2014.09.026.
31. 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 : 244–253. doi: 10.1016/j.freeradbiomed.2013.05.050.
32. Zhao F, Zhu J, Shi L et al. OGG1 in the kidney: beyond base excision repair. Oxid Med Cell Longev 2022; 2022 : 5774641. doi: 10.1155/2022/5774641.
33. Shigenaga MK, Gimeno CJ, Ames BN. Urinary 8-hydroxy-2’-deoxyguanosine as a biological marker of in vivo oxidative DNA damage. Proc Natl Acad Sci U S A 1989; 86 (24): 9697–9701. doi: 10.1073/pnas.86.24.9697.
34. Suzuki J, Inoue Y, Suzuki S. Changes in the urinary excretion level of 8-hydroxyguanine by exposure to reactive oxygen-generating substances. Free Radic Biol Med 1995; 18 (3): 431–436. doi: 10.1016/0891-5849 (94) 00152-A.
35. Moriwaki H. Determination of 8-hydroxy-2’-deoxyguanosine in urine by liquid chromatography–electrospray ionization–mass spectrometry. Anal Sci 2000; 16 (2): 105–106. doi: 10.2116/analsci.16.105.
36. Kadiiska MB, Gladen BC, Baird DD et al. Biomarkers of oxidative stress study II. Are oxidation products of lipids, proteins, and DNA markers of CCl4 poisoning? Free Radic Biol Med 2005; 38 (5): 698–710. doi: 10.1016/j.freeradbiomed.2004.09.017.
37. Il’yasova D, Scarbrough P, Spasojevic I. Urinary biomarkers of oxidative status. Clin Chim Acta 2012; 413 (19–20): 1446–1453. doi: 10.1016/j.cca.2012.06.012.
38. Kim K. Glutathione in the nervous system as a potential therapeutic target to control the development and progression of amyotrophic lateral sclerosis. Antioxidants (Basel) 2021; 10 (7): 1011. doi: 10.3390/antiox10071011.
39. Dringen R, Arend C. Glutathione metabolism of the brain – the role of astrocytes. J Neurochem 2025; 169 (5): e70073. doi: 10.1111/jnc.70073.
40. Vargas MR, Johnson JA. The Nrf2-ARE cytoprotective pathway in astrocytes. Expert Rev Mol Med 2009; 11: e17. doi: 10.1017/S1462399409001094.
41. Zhang W, Li P, Geng Q et al. Simultaneous determination of glutathione, cysteine, homocysteine, and cysteinylglycine in biological fluids by ion-pairing high-performance liquid chromatography coupled with precolumn derivatization. J Agric Food Chem 2014; 62 (25): 5845–5852. doi: 10.1021/jf5014007.
42. Weiduschat N, Mao X, Hupf J et al. Motor cortex glutathione deficit in ALS measured in vivo with the J-editing technique. Neurosci Lett 2014; 570 : 102–107. doi: 10.1016/j.neulet.2014.04.020.
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Paediatric neurology Neurosurgery NeurologyArticle was published in
Czech and Slovak Neurology and Neurosurgery
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