Abstract
Thioredoxin (Trx) and glyoxalase (Glo) systems have been suggested to be molecular targets of methylglyoxal (MGO). This highly reactive endogenous compound has been associated with the development of neurodegenerative pathologies and cell death. In the present study, the glutathione (GSH), Trx, and Glo systems were investigated to understand early events (0.5–3 h) that may determine cell fate. It is shown for the first time that MGO treatment induces an increase in glutathione reductase (GR) protein in hippocampal slices (1 h) and HT22 nerve cells (0.5 and 2.5 h). Thioredoxin interacting protein (Txnip), thioredoxin reductase (TrxR), Glo1, and Glo2 were markedly increased (2- to 4-fold) in hippocampal slices and 1.2- to 1.3-fold in HT22 cells. This increase in protein levels in hippocampal slices was followed by a corresponding increase in GR, TrxR, and Glo1 activities, but not in HT22 cells. In these cells, GR and TrxR activities were decreased by MGO. This result is in agreement with the idea that MGO can affect the Trx/TrxR reducing system, and now we show that GR and Txnip can also be affected by MGO. Impairment in the GR or TrxR reducing capacity can impair peroxide removal by glutathione peroxidase and peroxiredoxin, as both peroxidases depend on reduced GSH and Trx, respectively. In this regard, inhibition of GR and TrxR by 2-AAPA or auranofin, respectively, potentiated MGO toxicity in differentiated SH-SY5Y cells. Overall, MGO not only triggers a clear defense response in hippocampal slices and HT22 cells but also impairs the Trx/TrxR and GSH/GR reducing couples in HT22 cells. The increased MGO toxicity caused by inhibition of GR and TrxR with specific inhibitors, or their inhibition by MGO treatment, supports the notion that both reducing systems are relevant molecular targets of MGO.
Similar content being viewed by others
References
Allaman I, Bélanger M, Magistretti PJ (2015) Methylglyoxal, the dark side of glycolysis. Front Neurosci. doi:10.3389/fnins.2015.00023
Angeloni C, Zambonin L, Hrelia S (2014) Role of methylglyoxal in Alzheimer’s disease. Biomed Res Int 2014:e238485. doi:10.1155/2014/238485
Arnér ES, Zhong L, Holmgren A (1999) Preparation and assay of mammalian thioredoxin and thioredoxin reductase. Methods Enzymol 300:226–239
Arscott LD, Veine DM, Williams CH (2000) Mixed disulfide with glutathione as an intermediate in the reaction catalyzed by glutathione reductase from yeast and as a major form of the enzyme in the cell. Biochemistry (Mosc) 39:4711–4721. doi:10.1021/bi9926431
Bevensee MO, Schwiening CJ, Boron WF (1995) Use of BCECF and propidium iodide to assess membrane integrity of acutely isolated CA1 neurons from rat hippocampus. J Neurosci Methods 58:61–75
Bishop GM, Dringen R, Robinson SR (2007) Zinc stimulates the production of toxic reactive oxygen species (ROS) and inhibits glutathione reductase in astrocytes. Free Radic Biol Med 42:1222–1230. doi:10.1016/j.freeradbiomed.2007.01.022
Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254
Carlberg I, Mannervik B (1985) Glutathione reductase. Methods Enzymol 113:484–490
Chang Y-C, Hsieh M-C, Wu H-J, Wu W-C, Kao Y-H (2015) Methylglyoxal, a reactive glucose metabolite, enhances autophagy flux and suppresses proliferation of human retinal pigment epithelial ARPE-19 cells. Toxicol in Vitro 29:1358–1368. doi:10.1016/j.tiv.2015.05.014
Chou H-C, Chan H-L (2014) Effect of glutathione reductase knockdown in response to UVB-induced oxidative stress in human lung adenocarcinoma. Proteome Sci 12:2. doi:10.1186/1477-5956-12-2
Cordova FM, Rodrigues ALS, Giacomelli MBO, Oliveira CS, Posser T, Dunkley PR, Leal RB (2004) Lead stimulates ERK1/2 and p38MAPK phosphorylation in the hippocampus of immature rats. Brain Res 998:65–72. doi:10.1016/j.brainres.2003.11.012
Currais A, Farrokhi C, Dargusch R, Goujon-Svrzic M, Maher P (2016) Dietary glycemic index modulates the behavioral and biochemical abnormalities associated with autism spectrum disorder. Mol Psychiatry 21:426–436. doi:10.1038/mp.2015.64
Dafre AL, Goldberg J, Wang T, Spiegel DA, Maher P (2015) Methylglyoxal, the foe and friend of glyoxalase and Trx/TrxR systems in HT22 nerve cells. Free Radic Biol Med 89:8–19. doi:10.1016/j.freeradbiomed.2015.07.005
Dafre AL, Schmitz AE, Maher P (2017) Methylglyoxal-induced AMPK activation leads to autophagic degradation of thioredoxin 1 and glyoxalase 2 in HT22 nerve cells. Free Radic Biol Med 108:270–279
Desai KM, Chang T, Wang H, Banigesh A, Dhar A, Liu J, Untereiner A, Wu L (2010) Oxidative stress and aging: is methylglyoxal the hidden enemy? Can J Physiol Pharmacol 88:273–284. doi:10.1139/Y10-001
Distler MG, Plant LD, Sokoloff G, Hawk AJ, Aneas I, Wuenschell GE, Termini J, Meredith SC, Nobrega MA, Palmer AA (2012) Glyoxalase 1 increases anxiety by reducing GABAA receptor agonist methylglyoxal. J Clin Invest 122:2306–2315. doi:10.1172/JCI61319
Drechsel DA, Patel M (2010) Respiration-dependent H2O2 removal in brain mitochondria via the thioredoxin/peroxiredoxin system. J Biol Chem 285:27850–27858. doi:10.1074/jbc.M110.101196
Dringen R (2000) Metabolism and functions of glutathione in brain. Prog Neurobiol 62:649–671. doi:10.1016/S0301-0082(99)00060-X
Dringen R, Pawlowski PG, Hirrlinger J (2005) Peroxide detoxification by brain cells. J Neurosci Res 79:157–165. doi:10.1002/jnr.20280
Dringen R, Brandmann M, Hohnholt MC, Blumrich E-M (2015) Glutathione-dependent detoxification processes in astrocytes. Neurochem Res 40:2570–2582. doi:10.1007/s11064-014-1481-1
Ehren JL, Maher P (2013) Concurrent regulation of the transcription factors Nrf2 and ATF4 mediates the enhancement of glutathione levels by the flavonoid fisetin. Biochem Pharmacol 85:1816–1826. doi:10.1016/j.bcp.2013.04.010
Gassmann M, Grenacher B, Rohde B, Vogel J (2009) Quantifying Western blots: pitfalls of densitometry. Electrophoresis 30:1845–1855. doi:10.1002/elps.200800720
Gilda JE, Gomes AV (2013) Stain free total protein staining is a superior loading control to β-actin for Western blots. Anal Biochem. doi:10.1016/j.ab.2013.05.027
Guerrero-Beltrán CE, Calderón-Oliver M, Pedraza-Chaverri J, Chirino YI (2012) Protective effect of sulforaphane against oxidative stress: recent advances. Exp Toxicol Pathol 64:503–508. doi:10.1016/j.etp.2010.11.005
Hambsch B (2011) Altered glyoxalase 1 expression in psychiatric disorders: cause or consequence? Semin Cell Dev Biol 22:302–308. doi:10.1016/j.semcdb.2011.02.005
Hawkes H-JK, Karlenius TC, Tonissen KF (2014) Regulation of the human thioredoxin gene promoter and its key substrates: a study of functional and putative regulatory elements. Biochim Biophys Acta BBA - Gen Subj 1840:303–314. doi:10.1016/j.bbagen.2013.09.013
Kalapos MP (2008) The tandem of free radicals and methylglyoxal. Chem Biol Interact 171:251–271. doi:10.1016/j.cbi.2007.11.009
Kalapos MP (2013) Where does plasma methylglyoxal originate from? Diabetes Res Clin Pract 99:260–271. doi:10.1016/j.diabres.2012.11.003
Kikuchi S, Shinpo K, Moriwaka F, Makita Z, Miyata T, Tashiro K (1999) Neurotoxicity of methylglyoxal and 3-deoxyglucosone on cultured cortical neurons: synergism between glycation and oxidative stress, possibly involved in neurodegenerative diseases. J Neurosci Res 57:280–289. doi:10.1002/(SICI)1097-4547(19990715)57:2<280::AID-JNR14>3.0.CO;2-U
Kimura R, Okouchi M, Fujioka H, Ichiyanagi A, Ryuge F, Mizuno T, Imaeda K, Okayama N, Kamiya Y, Asai K, Joh T (2009) Glucagon-like peptide-1 (GLP-1) protects against methylglyoxal-induced PC12 cell apoptosis through the PI3K/Akt/mTOR/GCLc/redox signaling pathway. Neurosci 162:1212–1219. doi:10.1016/j.neuroscience.2009.05.025
Kuhla B, Lüth H-J, Haferburg D, Boeck K, Arendt T, Münch G (2005) Methylglyoxal, glyoxal, and their detoxification in Alzheimer’s disease. Ann N Y Acad Sci 1043:211–216. doi:10.1196/annals.1333.026
Lennicke C, Rahn J, Lichtenfels R, Wessjohann LA, Seliger B (2015) Hydrogen peroxide—production, fate and role in redox signaling of tumor cells. Cell Commun Signal CCS. doi:10.1186/s12964-015-0118-6
Liu Y, Peterson DA, Kimura H, Schubert D (1997) Mechanism of cellular 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) reduction. J Neurochem 69:581–593. doi:10.1046/j.1471-4159.1997.69020581.x
Liu H, Yu S, Zhang H, Xu J (2012) Angiogenesis impairment in diabetes: role of methylglyoxal-induced receptor for advanced glycation endproducts, autophagy and vascular endothelial growth factor receptor 2. PLoS One 7:e46720. doi:10.1371/journal.pone.0046720
Lopes FM, Schröder R, da Frota ML Jr, Zanotto-Filho A, Müller CB, Pires AS, Meurer RT, Colpo GD, Gelain DP, Kapczinski F, JCF M, Fernandes Mda C, Klamt F (2010) Comparison between proliferative and neuron-like SH-SY5Y cells as an in vitro model for Parkinson disease studies. Brain Res 1337:85–94. doi:10.1016/j.brainres.2010.03.102
Lopes FM, da Motta LL, Bastiani MAD, Pfaffenseller B, Aguiar BW, de Souza LF, Zanatta G, Vargas DM, Schönhofen P, Londero GF, de Medeiros LM, Freire VN, Dafre AL, Castro MAA, Parsons RB, Klamt F (2017) RA differentiation enhances dopaminergic features, changes redox parameters, and increases dopamine transporter dependency in 6-hydroxydopamine-induced neurotoxicity in SH-SY5Y cells. Neurotox Res 1–15. doi:10.1007/s12640-016-9699-0
Lu J, Holmgren A (2014) The thioredoxin antioxidant system. Free Radic Biol Med 66:75–87. doi:10.1016/j.freeradbiomed.2013.07.036
Lubos E, Loscalzo J, Handy DE (2010) Glutathione peroxidase-1 in health and disease: from molecular mechanisms to therapeutic opportunities. Antioxid Redox Signal 15:1957–1997. doi:10.1089/ars.2010.3586
Maher P (2012) Methylglyoxal, advanced glycation end products and autism: is there a connection? Med Hypotheses 78:548–552. doi:10.1016/j.mehy.2012.01.032
Matafome P, Sena C, Seiça R (2013) Methylglyoxal, obesity, and diabetes. Endocrine 43:472–484. doi:10.1007/s12020-012-9795-8
Mitozo PA, De Souza LF, Loch-Neckel G, Flesch S, Maris AF, Figueiredo CP, Dos Santos ARS, Farina M, Dafre AL (2011) A study of the relative importance of the peroxiredoxin-, catalase-, and glutathione-dependent systems in neural peroxide metabolism. Free Radic Biol Med 51:69–77. doi:10.1016/j.freeradbiomed.2011.03.017
Molz S, Decker H, Dal-Cim T, Cremonez C, Cordova FM, Leal RB, Tasca CI (2008) Glutamate-induced toxicity in hippocampal slices involves apoptotic features and p38MAPK signaling. Neurochem Res 33:27–36. doi:10.1007/s11064-007-9402-1
Oba T, Tatsunami R, Sato K, Takahashi K, Hao Z, Tampo Y (2012) Methylglyoxal has deleterious effects on thioredoxin in human aortic endothelial cells. Environ Toxicol Pharmacol 34:117–126. doi:10.1016/j.etap.2012.03.007
Okouchi M, Okayama N, Aw T (2005) Hyperglycemia potentiates carbonyl stress-induced apoptosis in naive PC-12 cells: relationship to cellular redox and activator protease factor-1 expression. Curr Neurovasc Res 2:375–386. doi:10.2174/156720205774962665
Övey İS, Naziroğlu M (2015) Homocysteine and cytosolic GSH depletion induce apoptosis and oxidative toxicity through cytosolic calcium overload in the hippocampus of aged mice: involvement of TRPM2 and TRPV1 channels. Neuroscience 284:225–233. doi:10.1016/j.neuroscience.2014.09.078
Perkins A, Nelson KJ, Parsonage D, Poole LB, Karplus PA (2015) Peroxiredoxins: guardians against oxidative stress and modulators of peroxide signaling. Trends Biochem Sci 40:435–445. doi:10.1016/j.tibs.2015.05.001
Poynton RA, Hampton MB (2014) Peroxiredoxins as biomarkers of oxidative stress. Biochim Biophys Acta BBA - Gen Subj 1840:906–912. doi:10.1016/j.bbagen.2013.08.001
Prior M, Chiruta C, Currais A, Goldberg J, Ramsey J, Dargusch R, Maher PA, Schubert D (2014) Back to the future with phenotypic screening. ACS Chem Neurosci 5:503–513. doi:10.1021/cn500051h
Qiao S, Dennis M, Song X, Vadysirisack DD, Salunke D, Nash Z, Yang Z, Liesa M, Yoshioka J, Matsuzawa S-I, Shirihai OS, Lee RT, Reed JC, Ellisen LW (2015) A REDD1/TXNIP pro-oxidant complex regulates ATG4B activity to control stress-induced autophagy and sustain exercise capacity. Nat Commun 6:7014. doi:10.1038/ncomms8014
Rabbani N, Xue M, Thornalley PJ (2016a) Methylglyoxal-induced dicarbonyl stress in aging and disease: first steps towards glyoxalase 1-based treatments. Clin Sci 130:1677–1696. doi:10.1042/CS20160025
Rabbani N, Xue M, Thornalley PJ (2016b) Dicarbonyls and glyoxalase in disease mechanisms and clinical therapeutics. Glycoconj J 33:513–525. doi:10.1007/s10719-016-9705-z
Racker E (1951) The mechanism of action of glyoxalase. J Biol Chem 190:685–696
Rodnight R, Gonçalves CA, Leal R, Rocha E, Salbego CG, Wofchuk ST (1991) Chapter 11: Regional distribution and properties of an enzyme system in rat brain that phosphorylates ppH-47, an insoluble protein highly labelled in tissue slices from the hippocampus. In: Routtenberg WHG and A (ed) Progress in Brain Research. Elsevier, pp 157–167
Saccoccia F, Angelucci F, Boumis G, Carotti D, Desiato G, Miele AE, Bellelli A (2014) Thioredoxin reductase and its inhibitors. Curr Protein Pept Sci 15:621–646. doi:10.2174/1389203715666140530091910
Schneider L, Giordano S, Zelickson BR, S Johnson M, A Benavides G, Ouyang X, Fineberg N, Darley-Usmar VM, Zhang J (2011) Differentiation of SH-SY5Y cells to a neuronal phenotype changes cellular bioenergetics and the response to oxidative stress. Free Radic Biol Med 51:2007–2017. doi:10.1016/j.freeradbiomed.2011.08.030
Selenius M, Rundlöf A-K, Olm E, Fernandes AP, Björnstedt M (2010) Selenium and the selenoprotein thioredoxin reductase in the prevention, treatment and diagnostics of cancer. Antioxid Redox Signal 12:867–880. doi:10.1089/ars.2009.2884
Shangari N, O’Brien PJ (2004) The cytotoxic mechanism of glyoxal involves oxidative stress. Biochem Pharmacol 68:1433–1442. doi:10.1016/j.bcp.2004.06.013
Shin MJ, Kim DW, Lee YP, Ahn EH, Jo HS, Kim D-S, Kwon O-S, Kang T-C, Cho Y-J, Park J, Eum WS, Choi SY (2014) Tat-glyoxalase protein inhibits against ischemic neuronal cell damage and ameliorates ischemic injury. Free Radic Biol Med 67:195–210. doi:10.1016/j.freeradbiomed.2013.10.815
Tatsunami R, Oba T, Takahashi K, Tampo Y (2009) Methylglyoxal causes dysfunction of thioredoxin and thioredoxin reductase in endothelial cells. J Pharmacol Sci 111:426–432. doi:10.1254/jphs.09131FP
Thornalley PJ (1996) Pharmacology of methylglyoxal: formation, modification of proteins and nucleic acids, and enzymatic detoxification—a role in pathogenesis and antiproliferative chemotherapy. Gen Pharmacol 27:565–573
Thornalley PJ (2003) Glyoxalase I-structure, function and a critical role in the enzymatic defence against glycation. Biochem Soc Trans 31:1343–1348
Thornalley PJ, Rabbani N (2011) Glyoxalase in tumourigenesis and multidrug resistance. Semin Cell Dev Biol 22:318–325. doi:10.1016/j.semcdb.2011.02.006
Thorne D, Kilford J, Payne R, Haswell L, Dalrymple A, Meredith C, Dillon D (2014) Development of a BALB/c 3T3 neutral red uptake cytotoxicity test using a mainstream cigarette smoke exposure system. BMC Res Notes 7:367. doi:10.1186/1756-0500-7-367
Tietze F (1969) Enzymic method for quantitative determination of nanogram amounts of total and oxidized glutathione: applications to mammalian blood and other tissues. Anal Biochem 27:502–522. doi:10.1016/0003-2697(69)90064-5
Vander Jagt DL, Hunsaker LA, Vander Jagt TJ, Gomez MS, Gonzales DM, Deck LM, Royer RE (1997) Inactivation of glutathione reductase by 4-hydroxynonenal and other endogenous aldehydes. Biochem Pharmacol 53:1133–1140. doi:10.1016/S0006-2952(97)00090-7
Wang X, Desai K, Chang T, Wu L (2005) Vascular methylglyoxal metabolism and the development of hypertension. J Hypertens 23:1565–1573
Wang X-L, Lau WB, Yuan Y-X, Wang Y-J, Yi W, Christopher TA, Lopez BL, Liu H-R, Ma X-L (2010) Methylglyoxal increases cardiomyocyte ischemia-reperfusion injury via glycative inhibition of thioredoxin activity. Am J Physiol - Endocrinol Metab 299:E207–E214. doi:10.1152/ajpendo.00215.2010
Wu L, Juurlink BHJ (2002) Increased methylglyoxal and oxidative stress in hypertensive rat vascular smooth muscle cells. Hypertens Dallas Tex 1979 39:809–814
Xue M, Rabbani N, Momiji H, Imbasi P, Anwar MM, Kitteringham N, Park BK, Souma T, Moriguchi T, Yamamoto M, Thornalley PJ (2012) Transcriptional control of glyoxalase 1 by Nrf2 provides a stress-responsive defence against dicarbonyl glycation. Biochem J 443:213–222. doi:10.1042/BJ20111648
Yoshihara E, Masaki S, Matsuo Y, Chen Z, Tian H, Yodoi J (2014) Thioredoxin/Txnip: redoxisome, as a redox switch for the pathogenesis of diseases. Inflammation 4:514. doi:10.3389/fimmu.2013.00514
Acknowledgements
This work was supported by the Brazilian funding agencies Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq; #462333/2014-0, #306204/2014-2) and a Thome Foundation grant to P. Maher. Luiz Felipe de Souza and Ariana Ern Schmitz were CAPES (Coordination for the Improvement of Higher Education Personnel) Ph.D. fellows. Alcir L. Dafre is a CNPq productivity research fellow.
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
All experiments were carried out in accordance with the guidelines on animal care of the local Ethics Committee on the use of animals (CEUA/UFSC), which follows the NIH publication “Principles of Laboratory Animal Care.”
Conflict of Interest
The authors declare that there are no conflicts of interest.
Rights and permissions
About this article
Cite this article
Schmitz, A.E., de Souza, L.F., dos Santos, B. et al. Methylglyoxal-Induced Protection Response and Toxicity: Role of Glutathione Reductase and Thioredoxin Systems. Neurotox Res 32, 340–350 (2017). https://doi.org/10.1007/s12640-017-9738-5
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s12640-017-9738-5