{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,5,5]],"date-time":"2026-05-05T06:17:43Z","timestamp":1777961863309,"version":"3.51.4"},"reference-count":51,"publisher":"MDPI AG","issue":"16","license":[{"start":{"date-parts":[[2025,8,12]],"date-time":"2025-08-12T00:00:00Z","timestamp":1754956800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"GUHEPRS and GUIPRS provided by Griffith University"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Molecules"],"abstract":"<jats:p>This study used NMR-based metabolomics to investigate the mode of action (MoA) of 6-hydroxydopamine (6-OHDA) toxicity in the SH-SY5Y neuroblastoma cell model. 6-OHDA, a structural analogue of dopamine, has been used to create a Parkinson\u2019s disease model since 1968. Its selective uptake via catecholaminergic transporters leads to intracellular oxidative stress and mitochondrial dysfunction. SH-SY5Y cells were treated with 6-OHDA at its IC50 concentration of 60 \u03bcM, and samples of treated and untreated groups were collected after 24 h. The endo metabolome was extracted using a methanol\u2013water mixture, while the exo metabolome was represented by the culture media. Further, endo- and exo metabolomes of treated and untreated cells were analysed for metabolic changes. Our results demonstrated significantly high levels of glutathione, acetate, propionate, and NAD+, which are oxidative stress markers, enhanced due to ROS production in the system. In addition, alteration of myoinositol, taurine, and o-phosphocholine could be due to oxidative stress-induced membrane potential disturbance. Mitochondrial complex I inhibition causes electron transport chain (ETC) dysfunction. Changes in key metabolites of glycolysis and energy metabolism, such as glucose, pyruvate, lactate, creatine, creatine phosphate, glycine, and methionine, respectively, demonstrated ETC dysfunction. We also identified changes in amino acids such as glutamine, glutamate, and proline, followed by nucleotide metabolism such as uridine and uridine monophosphate levels, which were decreased in the treated group.<\/jats:p>","DOI":"10.3390\/molecules30163352","type":"journal-article","created":{"date-parts":[[2025,8,12]],"date-time":"2025-08-12T16:30:36Z","timestamp":1755016236000},"page":"3352","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["Mode of Action of Toxin 6-Hydroxydopamine in SH-SY5Y Using NMR Metabolomics"],"prefix":"10.3390","volume":"30","author":[{"given":"Roktima","family":"Tamuli","sequence":"first","affiliation":[{"name":"Institute for Biomedicine and Glycomics, Griffith University, Nathan, QLD 4111, Australia"},{"name":"School of Environment and Science, Griffith University, Nathan, QLD 4111, Australia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"George D.","family":"Mellick","sequence":"additional","affiliation":[{"name":"Institute for Biomedicine and Glycomics, Griffith University, Nathan, QLD 4111, Australia"},{"name":"School of Environment and Science, Griffith University, Nathan, QLD 4111, Australia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7541-246X","authenticated-orcid":false,"given":"Horst Joachim","family":"Schirra","sequence":"additional","affiliation":[{"name":"Institute for Biomedicine and Glycomics, Griffith University, Nathan, QLD 4111, Australia"},{"name":"School of Environment and Science, Griffith University, Nathan, QLD 4111, Australia"},{"name":"Centre for Advanced Imaging, The University of Queensland, Brisbane, QLD 4072, Australia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2412-1213","authenticated-orcid":false,"given":"Yunjiang","family":"Feng","sequence":"additional","affiliation":[{"name":"Institute for Biomedicine and Glycomics, Griffith University, Nathan, QLD 4111, Australia"},{"name":"School of Environment and Science, Griffith University, Nathan, QLD 4111, Australia"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2025,8,12]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"377","DOI":"10.1038\/nrd3430","article-title":"Priorities in Parkinson\u2019s disease research","volume":"10","author":"Meissner","year":"2011","journal-title":"Nat. Rev. Drug Discov."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"e464","DOI":"10.1016\/S2666-7568(24)00094-1","article-title":"Temporal trends in the prevalence of Parkinson\u2019s disease from 1980 to 2023: A systematic review and meta-analysis","volume":"5","author":"Zhu","year":"2024","journal-title":"Lancet Healthy Longev."},{"key":"ref_3","first-page":"533","article-title":"The 6-hydroxydopamine model and parkinsonian pathophysiology Novel findings in an older model","volume":"32","year":"2017","journal-title":"Neurolog\u00eda"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"1156","DOI":"10.1111\/j.1742-4658.2012.08491.x","article-title":"Animal models of Parkinson\u2019s disease","volume":"279","author":"Blandini","year":"2012","journal-title":"FEBS J."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"2114","DOI":"10.1021\/acs.jproteome.2c00149","article-title":"Proteomic and Targeted Metabolomic Studies on a Silkworm Model of Parkinson\u2019s Disease","volume":"21","author":"Zhu","year":"2022","journal-title":"J. Proteome Res."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"151","DOI":"10.1007\/BF03033565","article-title":"The 6-Hydroxydopamine Model of Parkinson\u2019s Disease","volume":"11","author":"Simola","year":"2007","journal-title":"Neurotox. Res."},{"key":"ref_7","doi-asserted-by":"crossref","unstructured":"Kaczy\u0144ska, K., and Andrzejewski, K. (2020). 6-hydroxydopamine-induced model of Parkinson\u2019s disease. Genetics, Neurology, Behavior, and Diet in Parkinson\u2019s Disease, Academic Press.","DOI":"10.1016\/B978-0-12-815950-7.00040-0"},{"key":"ref_8","doi-asserted-by":"crossref","unstructured":"Kostrzewa, R.M. (2014). 6-Hydroxydopamine as Preclinical Model of Parkinson\u2019s Disease. Handbook of Neurotoxicity, Springer.","DOI":"10.1007\/978-1-4614-5836-4"},{"key":"ref_9","doi-asserted-by":"crossref","unstructured":"Glinka, Y., Gassen, M., and Youdim, M.B.H. (1997). Mechanism of 6-Hydroxydopamine Neurotoxicity, Springer.","DOI":"10.1007\/978-3-7091-6842-4_7"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"675","DOI":"10.1016\/j.freeradbiomed.2006.12.004","article-title":"Molecular mechanisms of 6-hydroxydopamine-induced cytotoxicity in PC12 cells: Involvement of hydrogen peroxide-dependent and -independent action","volume":"42","author":"Saito","year":"2007","journal-title":"Free Radic. Biol. Med."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"17","DOI":"10.1186\/1750-1326-9-17","article-title":"The Parkinsonian mimetic, 6-OHDA, impairs axonal transport in dopaminergic axons","volume":"9","author":"Lu","year":"2014","journal-title":"Mol. Neurodegener."},{"key":"ref_12","first-page":"329","article-title":"Inhibition of mitochondrial complexes I and IV by 6-hydroxydopamine","volume":"292","author":"Glinka","year":"1995","journal-title":"Eur. J. Pharmacol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"135","DOI":"10.1016\/S0301-0082(01)00003-X","article-title":"Molecular pathways involved in the neurotoxicity of 6-OHDA, dopamine and MPTP-contribution to the apoptotic theory in Parkinson\u2019s disease","volume":"65","author":"Blum","year":"2001","journal-title":"Prog. Neurobiol."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"848","DOI":"10.1007\/s12035-019-01733-3","article-title":"Lipid Analysis of the 6-Hydroxydopamine-Treated SH-SY5Y Cell Model for Parkinson\u2019s Disease","volume":"57","author":"Xicoy","year":"2020","journal-title":"Mol. Neurobiol."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"20","DOI":"10.31083\/j.jin2201020","article-title":"SH-SY5Y Cell Line In Vitro Models for Parkinson Disease Research-Old Practice for New Trends","volume":"22","author":"Ioghen","year":"2023","journal-title":"J. Integr. Neurosci."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"10","DOI":"10.1186\/s13024-017-0149-0","article-title":"The SH-SY5Y cell line in Parkinson\u2019s disease research: A systematic review","volume":"12","author":"Xicoy","year":"2017","journal-title":"Mol. Neurodegener."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"969","DOI":"10.1039\/b909989c","article-title":"Natural product mode of action (MOA) studies: A link between natural and synthetic worlds","volume":"27","year":"2010","journal-title":"Nat. Prod. Rep."},{"key":"ref_18","doi-asserted-by":"crossref","unstructured":"Moco, S. (2022). Studying Metabolism by NMR-Based Metabolomics. Front. Mol. Biosci., 9.","DOI":"10.3389\/fmolb.2022.882487"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"19","DOI":"10.1007\/978-3-030-51652-9_2","article-title":"NMR-Based Metabolomics","volume":"1280","author":"Raftery","year":"2021","journal-title":"Adv. Exp. Med. Biol."},{"key":"ref_20","doi-asserted-by":"crossref","unstructured":"Alwahsh, M., Nimer, R.M., Dahabiyeh, L.A., Hamadneh, L., Hasan, A., Alejel, R., and Hergenroder, R. (2024). NMR-based metabolomics identification of potential serum biomarkers of disease progression in patients with multiple sclerosis. Sci. Rep., 14.","DOI":"10.1038\/s41598-024-64490-x"},{"key":"ref_21","doi-asserted-by":"crossref","unstructured":"Gowda, G.A.N., and Raftery, D. (2019). NMR-Based Metabolomics Methods and Protocols, Springer Protocols.","DOI":"10.1007\/978-1-4939-9690-2"},{"key":"ref_22","doi-asserted-by":"crossref","unstructured":"Seymour, G.J., Cullinan, M.P., and Heng, N.C.K. (2017). NMR-Based Metabolomics of Oral Biofluids. Oral Biology: Molecular Techniques and Applications, Springer.","DOI":"10.1007\/978-1-4939-6685-1"},{"key":"ref_23","unstructured":"Gowda, G.A.N., and Raftery, D. (2019). Practical Aspects of the Analysis of Low- and High-Field NMR Data from Environmental Samples. NMR-Based Metabolomics: Methods and Protocols, Springer."},{"key":"ref_24","doi-asserted-by":"crossref","unstructured":"Gowda, G.A.N., and Raftery, D. (2019). Plant Metabolomics Using NMR Spectroscopy. NMR-Based Metabolomics: Methods and Protocols, Springer.","DOI":"10.1007\/978-1-4939-9690-2"},{"key":"ref_25","doi-asserted-by":"crossref","unstructured":"Gowda, G.A.N., and Raftery, D. (2019). NMR-Based Metabolomics of Food. NMR-Based Metabolomics: Methods and Protocols, Springer.","DOI":"10.1007\/978-1-4939-9690-2"},{"key":"ref_26","doi-asserted-by":"crossref","unstructured":"Gowda, G.A.N., and Raftery, D. (2019). NMR Metabolomics Protocols for Drug Discovery. NMR-Based Metabolomics: Methods and Protocols, Springer.","DOI":"10.1007\/978-1-4939-9690-2"},{"key":"ref_27","doi-asserted-by":"crossref","unstructured":"Ji, J., Sun, J., Zhang, Y., and Sun, X. (2022). Cell-Based Metabolomics Approach for Anticipating and Investigating Cytotoxicity of Gold Nanorods. Foods, 11.","DOI":"10.3390\/foods11223569"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"2762","DOI":"10.1002\/elps.201200605","article-title":"Mammalian cell metabolomics: Experimental design and sample preparation","volume":"34","author":"Leon","year":"2013","journal-title":"Electrophoresis"},{"key":"ref_29","doi-asserted-by":"crossref","unstructured":"Sun, Y., Lu, Y., Joseph, C.M.L., Ma, L., Bisson, L.F., and Liu, Y. (2024). Metabolomic analysis reveals the interactions between Chinese indigenous and commercial Saccharomyces cerevisiae strains during wine co-fermentations at low YAN concentration. Food Bioscience, 60.","DOI":"10.1016\/j.fbio.2024.104362"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"W398","DOI":"10.1093\/nar\/gkae253","article-title":"MetaboAnalyst 6.0: Towards a unified platform for metabolomics data processing, analysis and interpretation","volume":"52","author":"Pang","year":"2024","journal-title":"Nucleic Acids Res."},{"key":"ref_31","unstructured":"Gupta, A., Gadellaa, M.M., and Maes, D.Y.W. (2011). Culture medium for eukaryotic cells. (EP2611904A2)."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"1406","DOI":"10.1016\/j.bbabio.2006.05.007","article-title":"Mitochondrial Complex I: Structural and functional aspects","volume":"1757","author":"Lenaz","year":"2006","journal-title":"Biochim. Biophys. Acta"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"335","DOI":"10.1016\/S0165-0173(97)00045-3","article-title":"Neurodegenerative disorders in humans: The role of glutathione in oxidative stress-mediated neuronal death","volume":"25","author":"Bains","year":"1997","journal-title":"Brain Res. Rev."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"271","DOI":"10.1038\/sj.embor.7400645","article-title":"The role of glutathione in disulphide bond formation and endoplasmic-reticulum-generated oxidative stress","volume":"7","author":"Chakravarthi","year":"2006","journal-title":"EMBO Rep."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"4904","DOI":"10.1046\/j.1432-1327.2000.01595.x","article-title":"Glutathione, oxidative stress and neurodegeneration","volume":"267","author":"Schulz","year":"2000","journal-title":"Eur. J. Biochem."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"385","DOI":"10.1016\/S1353-8020(02)00018-4","article-title":"Glutathione depletion and oxidative stress","volume":"8","author":"Mytilineou","year":"2002","journal-title":"Park. Relat. Disord."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"28","DOI":"10.1179\/1351000212Y.0000000001","article-title":"NAD+ metabolism and oxidative stress: The golden nucleotide on a crown of thorns","volume":"17","author":"Massudi","year":"2012","journal-title":"Redox Rep."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"502","DOI":"10.1016\/j.cell.2018.08.040","article-title":"Acetate Production from Glucose and Coupling to Mitochondrial Metabolism in Mammals","volume":"175","author":"Liu","year":"2018","journal-title":"Cell"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"10","DOI":"10.1016\/j.ejphar.2018.10.025","article-title":"The effects of jolkinolide B on HepG2 cells as revealed by (1)H-NMR-based metabolic profiling","volume":"842","author":"Yan","year":"2019","journal-title":"Eur. J. Pharmacol."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"868","DOI":"10.1021\/acs.biochem.2c00064","article-title":"Phosphonate and alpha-Fluorophosphonate Analogues of d-Glucose 6-Phosphate as Active-Site Probes of 1l-myo-Inositol 1-Phosphate Synthase","volume":"61","author":"Palmer","year":"2022","journal-title":"Biochemistry"},{"key":"ref_41","doi-asserted-by":"crossref","unstructured":"Chandel, N.S. (2021). Glycolysis. Cold Spring Harb. Perspect. Biol., 13.","DOI":"10.1101\/cshperspect.a040535"},{"key":"ref_42","doi-asserted-by":"crossref","unstructured":"Jones, B., Balasubramaniam, M., Lebowitz, J.J., Taylor, A., Villalta, F., Khoshbouei, H., Grueter, C., Grueter, B., Dash, C., and Pandhare, J. (2021). Activation of proline biosynthesis is critical to maintain glutamate homeostasis during acute methamphetamine exposure. Sci. Rep., 11.","DOI":"10.1038\/s41598-020-80917-7"},{"key":"ref_43","doi-asserted-by":"crossref","unstructured":"Cappelletti, P., Tallarita, E., Rabattoni, V., Campomenosi, P., Sacchi, S., and Pollegioni, L. (2018). Proline oxidase controls proline, glutamate, and glutamine cellular concentrations in a U87 glioblastoma cell line. PLoS ONE, 13.","DOI":"10.1371\/journal.pone.0196283"},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"384","DOI":"10.1016\/j.bbabio.2008.11.003","article-title":"Differential effects of mitochondrial Complex I inhibitors on production of reactive oxygen species","volume":"1787","author":"Fato","year":"2009","journal-title":"Biochim. Biophys. Acta"},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"331","DOI":"10.1016\/j.expneurol.2010.03.028","article-title":"Complex I-Inhibitors, inhibition and neurodegeneration","volume":"224","author":"Schapira","year":"2010","journal-title":"Exp. Neurol."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"351","DOI":"10.1126\/science.ade3332","article-title":"Structural basis of mammalian respiratory complex I inhibition by medicinal biguanides","volume":"379","author":"Bridges","year":"2023","journal-title":"Science"},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"3","DOI":"10.1016\/bs.mcb.2019.10.002","article-title":"Isolation of mitochondria from cells and tissues","volume":"155","author":"Liao","year":"2020","journal-title":"Methods Cell Biol."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"19","DOI":"10.1016\/S0895-4356(98)00138-3","article-title":"Stratified Randomization for Clinical Trials","volume":"52","author":"Kernan","year":"1999","journal-title":"Clin. Epidemiol."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"517","DOI":"10.1021\/ac048803i","article-title":"Evaluation of the orthogonal projection on latent structure model limitations caused by chemical shift variability and improved visualization of biomarker changes in 1H NMR spectroscopic setabonomic studies","volume":"77","author":"Cloarec","year":"2005","journal-title":"Anal. Chem."},{"key":"ref_50","doi-asserted-by":"crossref","unstructured":"Currin-Ross, D., Husdell, L., Pierens, G.K., Mok, N.E., O\u2019Neill, S.L., Schirra, H.J., and Brownlie, J.C. (2021). The Metabolic Response to Infection With Wolbachia Implicates the Insulin\/Insulin-Like-Growth Factor and Hypoxia Signaling Pathways in Drosophila melanogaster. Front. Ecol. Evol., 9.","DOI":"10.3389\/fevo.2021.623561"},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"1282","DOI":"10.1021\/ac048630x","article-title":"Statistical Total Correlation Spectroscopy: An Exploratory Approach for Latent Biomarker Identification from Metabolic 1 H NMR Data Sets","volume":"77","author":"Cloarec","year":"2005","journal-title":"Anal. Chem."}],"container-title":["Molecules"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1420-3049\/30\/16\/3352\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,9]],"date-time":"2025-10-09T18:25:30Z","timestamp":1760034330000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1420-3049\/30\/16\/3352"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,8,12]]},"references-count":51,"journal-issue":{"issue":"16","published-online":{"date-parts":[[2025,8]]}},"alternative-id":["molecules30163352"],"URL":"https:\/\/doi.org\/10.3390\/molecules30163352","relation":{},"ISSN":["1420-3049"],"issn-type":[{"value":"1420-3049","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,8,12]]}}}