{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T01:23:33Z","timestamp":1760059413891,"version":"build-2065373602"},"reference-count":92,"publisher":"MDPI AG","issue":"6","license":[{"start":{"date-parts":[[2025,6,12]],"date-time":"2025-06-12T00:00:00Z","timestamp":1749686400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100000780","name":"European Commission FP7 \u201cVetMedZg\u201d project","doi-asserted-by":"publisher","award":["621394","KK.01.1.1.04.0086"],"award-info":[{"award-number":["621394","KK.01.1.1.04.0086"]}],"id":[{"id":"10.13039\/501100000780","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100008530","name":"European Regional Development Fund","doi-asserted-by":"publisher","award":["621394","KK.01.1.1.04.0086"],"award-info":[{"award-number":["621394","KK.01.1.1.04.0086"]}],"id":[{"id":"10.13039\/501100008530","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Metabolites"],"abstract":"<jats:p>Background\/Objectives: Canine intervertebral disc herniation (IVDH) is an important musculoskeletal pathology. Unlike in humans, IVDH mechanisms in dogs are underinvestigated from a system-level integrative omics point of view. The aim of this study was to identify key serum molecular players in canine IVDH. Methods: An integrative multi-omics approach combining high-resolution LC-MS-based untargeted metabolomics and tandem mass tag (TMT)-based proteomics was applied. Additionally, serum zinc concentration was determined by spectrophotometry. Results: Nineteen serum metabolites were differentially abundant in IVDH dogs. Metabolite analysis highlighted dysregulation in lipoic acid and branched-chain amino acid (BCAA) metabolism, with elevated levels of valine, leucine, and isoleucine in IVDH. These findings suggest disrupted energy, nitrogen, and neurotransmitter metabolism, potentially contributing to the IVDH pathophysiology. Additionally, lower serum uridine, possibly influenced by BCAA accumulation, was observed, indicating altered neuroinflammatory responses. ELISA validation confirmed elevated serum levels of zinc-\u03b12-glycoprotein (ZAG), alpha-1-microglobulin\/bikunin precursor (AMBP), and vitronectin (VTN) in IVDH, supporting immune modulation and neuroprotective mechanisms. Serum prekallikrein (KLKB1) and Protein C inhibitor (SERPINA5), involved in fibrin cloth formation, were found to be lowered in IVDH patients. Pathway enrichment revealed disturbances in aromatic amino acid biosynthesis, with elevated phenylalanine, tyrosine, and tryptophan influencing neurotransmission and inflammation. In addition, elevated serum Zn concentration emphasized its antioxidant importance in immune response, wound healing, and neuropathic pain signaling. Conclusions: Integration with our prior CSF multi-omics data reinforced the relevance of identified molecules in IVDH-associated neurodegeneration, inflammation, and repair processes. This study offers insight into potential diagnostic biomarkers and therapeutic targets for canine IVDH through serum-based molecular profiling.<\/jats:p>","DOI":"10.3390\/metabo15060396","type":"journal-article","created":{"date-parts":[[2025,6,12]],"date-time":"2025-06-12T11:47:07Z","timestamp":1749728827000},"page":"396","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Combining Metabolomics and Proteomics to Reveal Key Serum Compounds Related to Canine Intervertebral Disc Herniation"],"prefix":"10.3390","volume":"15","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-9625-1450","authenticated-orcid":false,"given":"Anita","family":"Horvati\u0107","sequence":"first","affiliation":[{"name":"Faculty of Food Technology and Biotechnology, University of Zagreb, 10000 Zagreb, Croatia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Josipa","family":"Kule\u0161","sequence":"additional","affiliation":[{"name":"Faculty of Veterinary Medicine, University of Zagreb, 10000 Zagreb, Croatia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9195-646X","authenticated-orcid":false,"given":"Andrea","family":"Gelemanovi\u0107","sequence":"additional","affiliation":[{"name":"Mediterranean Institute for Life Sciences (MedILS), University of Split, 21000 Split, Croatia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ozren","family":"Smolec","sequence":"additional","affiliation":[{"name":"Faculty of Veterinary Medicine, University of Zagreb, 10000 Zagreb, Croatia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Boris","family":"Pirki\u0107","sequence":"additional","affiliation":[{"name":"Faculty of Veterinary Medicine, University of Zagreb, 10000 Zagreb, Croatia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1186-4774","authenticated-orcid":false,"given":"Marko","family":"Pe\u0107in","sequence":"additional","affiliation":[{"name":"Faculty of Veterinary Medicine, University of Zagreb, 10000 Zagreb, Croatia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ivana","family":"Rubi\u0107","sequence":"additional","affiliation":[{"name":"Faculty of Veterinary Medicine, University of Zagreb, 10000 Zagreb, Croatia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Vladimir","family":"Mrljak","sequence":"additional","affiliation":[{"name":"Faculty of Veterinary Medicine, University of Zagreb, 10000 Zagreb, Croatia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0402-3173","authenticated-orcid":false,"given":"Marko","family":"Samard\u017eija","sequence":"additional","affiliation":[{"name":"Faculty of Veterinary Medicine, University of Zagreb, 10000 Zagreb, Croatia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Marija","family":"Lipar","sequence":"additional","affiliation":[{"name":"Faculty of Veterinary Medicine, University of Zagreb, 10000 Zagreb, Croatia"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2025,6,12]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"2151","DOI":"10.1097\/01.brs.0000231761.73859.2c","article-title":"What Is Intervertebral Disc Degeneration, and What Causes It?","volume":"31","author":"Adams","year":"2006","journal-title":"Spine"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"e1109","DOI":"10.1002\/jsp2.1109","article-title":"Canine Models of Spine Disorders","volume":"3","author":"Lee","year":"2020","journal-title":"JOR Spine"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"1788","DOI":"10.3390\/neurolint16060129","article-title":"Animal Models of Intervertebral Disc Diseases: Advantages, Limitations, and Future Directions","volume":"16","author":"Hong","year":"2024","journal-title":"Neurol. Int."},{"key":"ref_4","doi-asserted-by":"crossref","unstructured":"da Costa, R.C., De Decker, S., Lewis, M.J., Volk, H., Moore, S.A., Olby, N.J., Levine, J.M., Lewis, M.J., Jeffery, N.D., and Mullins, M.E. (2020). Diagnostic Imaging in Intervertebral Disc Disease. Front. Vet. Sci., 7.","DOI":"10.3389\/fvets.2020.588338"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"116","DOI":"10.1002\/ca.23803","article-title":"Genetics of Intervertebral Disc Disease: A Review","volume":"35","author":"Ravichandran","year":"2022","journal-title":"Clin. Anat."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"675","DOI":"10.1089\/neu.2010.1645","article-title":"Naturally Occurring Disk Herniation in Dogs: An Opportunity for Pre-Clinical Spinal Cord Injury Research","volume":"28","author":"Levine","year":"2011","journal-title":"J. Neurotrauma"},{"key":"ref_7","doi-asserted-by":"crossref","unstructured":"Fenn, J., and Olby, N.J. (2020). Classification of Intervertebral Disc Disease. Front. Vet. Sci., 7.","DOI":"10.3389\/fvets.2020.579025"},{"key":"ref_8","unstructured":"Slatter, D.H. (2003). Textbook of Small Animal Surgery, Saunders. [3rd ed.]."},{"key":"ref_9","doi-asserted-by":"crossref","unstructured":"Liawrungrueang, W., Kim, P., Kotheeranurak, V., Jitpakdee, K., and Sarasombath, P. (2023). Automatic Detection, Classification, and Grading of Lumbar Intervertebral Disc Degeneration Using an Artificial Neural Network Model. Diagnostics, 13.","DOI":"10.3390\/diagnostics13040663"},{"key":"ref_10","doi-asserted-by":"crossref","unstructured":"Moore, S.A., Tipold, A., Olby, N.J., Stein, V., and Granger, N. (2020). Current Approaches to the Management of Acute Thoracolumbar Disc Extrusion in Dogs. Front. Vet. Sci., 7.","DOI":"10.3389\/fvets.2020.00610"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"158","DOI":"10.1038\/s41584-020-00568-w","article-title":"Cell-Based Strategies for IVD Repair: Clinical Progress and Translational Obstacles","volume":"17","author":"Binch","year":"2021","journal-title":"Nat. Rev. Rheumatol."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"113","DOI":"10.1021\/pr800545q","article-title":"Standard Operating Procedures for Serum and Plasma Collection: Early Detection Research Network Consensus Statement Standard Operating Procedure Integration Working Group","volume":"8","author":"Tuck","year":"2009","journal-title":"J. Proteome Res."},{"key":"ref_13","doi-asserted-by":"crossref","unstructured":"Jesuthasan, A., Ali, A., Lee, J.K.W., and Rutherfurd-Markwick, K. (2022). Assessment of Changes in Physiological Markers in Different Body Fluids at Rest and after Exercise. Nutrients, 14.","DOI":"10.3390\/nu14214685"},{"key":"ref_14","doi-asserted-by":"crossref","unstructured":"Horvatic, A., Gelemanovic, A., Pirkic, B., Smolec, O., Ljubic, B.B., Rubic, I., Eckersall, P.D., Mrljak, V., McLaughlin, M., and Samardzija, M. (2021). Multi-Omics Approach to Elucidate Cerebrospinal Fluid Changes in Dogs with Intervertebral Disc Herniation. Int. J. Mol. Sci., 22.","DOI":"10.3390\/ijms222111678"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"2036","DOI":"10.1039\/C6MB00220J","article-title":"Omics Approaches to Probe Markers of Disease Resistance in Animal Sciences","volume":"12","author":"Guillemin","year":"2016","journal-title":"Mol. Biosyst."},{"key":"ref_16","doi-asserted-by":"crossref","unstructured":"Schumacher-Schuh, A., Bieger, A., Borelli, W.V., Portley, M.K., Awad, P.S., and Bandres-Ciga, S. (2022). Advances in Proteomic and Metabolomic Profiling of Neurodegenerative Diseases. Front. Neurol., 12.","DOI":"10.3389\/fneur.2021.792227"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"503","DOI":"10.1016\/j.joca.2015.09.020","article-title":"Fibrotic-like Changes in Degenerate Human Intervertebral Discs Revealed by Quantitative Proteomic Analysis","volume":"24","author":"Yee","year":"2016","journal-title":"Osteoarthr. Cartil."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"376","DOI":"10.1016\/j.mri.2015.12.005","article-title":"Assessing the Correlation between the Degree of Disc Degeneration on the Pfirrmann Scale and the Metabolites Identified in HR-MAS NMR Spectroscopy","volume":"34","author":"Radek","year":"2016","journal-title":"Magn. Reson. Imaging"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"356","DOI":"10.4162\/nrp.2010.4.5.356","article-title":"Zinc May Increase Bone Formation through Stimulating Cell Proliferation, Alkaline Phosphatase Activity and Collagen Synthesis in Osteoblastic MC3T3-E1 Cells","volume":"4","author":"Seo","year":"2010","journal-title":"Nutr. Res. Pract."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"23","DOI":"10.1007\/s00204-024-03891-3","article-title":"Zinc and Its Binding Proteins: Essential Roles and Therapeutic Potential","volume":"99","author":"Kiouri","year":"2024","journal-title":"Arch. Toxicol."},{"key":"ref_21","doi-asserted-by":"crossref","unstructured":"Staszkiewicz, R., Bry\u015b, K., G\u0142adysz, D., Gralewski, M., Garczarek, M., Gadzieli\u0144ski, M., Wieczorek, J., Marcol, W., Ostenda, A., and Grabarek, B.O. (2022). Changes in Elements and Relationships among Elements in Intervertebral Disc Degeneration. Int. J. Environ. Res. Public Health, 19.","DOI":"10.3390\/ijerph19159042"},{"key":"ref_22","doi-asserted-by":"crossref","unstructured":"Pereira, A.M., Maia, M.R.G., Fonseca, A.J.M., and Cabrita, A.R.J. (2021). Zinc in Dog Nutrition, Health and Disease: A Review. Animals, 11.","DOI":"10.3390\/ani11040978"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"103997","DOI":"10.1016\/j.jprot.2020.103997","article-title":"Quantitative Proteomics of Cerebrospinal Fluid Using Tandem Mass Tags in Dogs with Recurrent Epileptic Seizures","volume":"231","author":"Baka","year":"2021","journal-title":"J. Proteom."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"4007","DOI":"10.1093\/bioinformatics\/btx499","article-title":"PiMP My Metabolome: An Integrated, Web-Based Tool for LC-MS Metabolomics Data","volume":"33","author":"Gloaguen","year":"2017","journal-title":"Bioinformatics"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"13.24.1","DOI":"10.1002\/0471250953.bi1324s46","article-title":"Employing ProteoWizard to Convert Raw Mass Spectrometry Data","volume":"46","author":"Holman","year":"2014","journal-title":"Curr. Protoc. Bioinform."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"64","DOI":"10.1016\/j.jprot.2018.08.009","article-title":"Quantitative Proteomics Using Tandem Mass Tags in Relation to the Acute Protein Response in Chicken Challenged with Escherichia coli Lipopolysaccharide Endotoxin","volume":"192","author":"Horvatic","year":"2019","journal-title":"J. Proteom."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"93","DOI":"10.1007\/s11306-016-1030-9","article-title":"Non-Targeted UHPLC-MS Metabolomic Data Processing Methods: A Comparative Investigation of Normalisation, Missing Value Imputation, Transformation and Scaling","volume":"12","author":"Engel","year":"2016","journal-title":"Metabolomics"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"W388","DOI":"10.1093\/nar\/gkab382","article-title":"MetaboAnalyst 5.0: Narrowing the Gap between Raw Spectra and Functional Insights","volume":"49","author":"Pang","year":"2021","journal-title":"Nucleic Acids Res."},{"key":"ref_29","unstructured":"Komsta, L. (2022, May 04). Outliers: Tests for Outliers; R Package Version 0.14; 2011. Available online: https:\/\/CRAN.R-project.org\/package=outliers."},{"key":"ref_30","first-page":"321","article-title":"About the Ggplot2 Package","volume":"5","author":"Wickham","year":"2016","journal-title":"J. Appl. Comput. Math."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"D607","DOI":"10.1093\/nar\/gky1131","article-title":"STRING V11: Protein\u2013Protein Association Networks with Increased Coverage, Supporting Functional Discovery in Genome-Wide Experimental Datasets","volume":"47","author":"Szklarczyk","year":"2019","journal-title":"Nucleic Acids Res."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"2498","DOI":"10.1101\/gr.1239303","article-title":"Cytoscape: A Software Environment for Integrated Models of Biomolecular Interaction Networks","volume":"13","author":"Shannon","year":"2003","journal-title":"Genome Res."},{"key":"ref_33","first-page":"100141","article-title":"ClusterProfiler 4.0: A Universal Enrichment Tool for Interpreting Omics Data","volume":"2","author":"Wu","year":"2021","journal-title":"Innovation"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"D583","DOI":"10.1093\/nar\/gkac831","article-title":"GotEnzymes: An Extensive Database of Enzyme Parameter Predictions","volume":"51","author":"Li","year":"2023","journal-title":"Nucleic Acids Res."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"e1230","DOI":"10.1002\/jsp2.1230","article-title":"An Update on Animal Models of Intervertebral Disc Degeneration and Low Back Pain: Exploring the Potential of Artificial Intelligence to Improve Research Analysis and Development of Prospective Therapeutics","volume":"6","author":"Alini","year":"2023","journal-title":"JOR Spine"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"1300","DOI":"10.2460\/javma.240.11.1300","article-title":"Incidence of intervertebral disk degeneration-related diseases and associated mortality rates in dogs","volume":"240","author":"eBergknut","year":"2012","journal-title":"J. Am. Vet. Med. Assoc."},{"key":"ref_37","doi-asserted-by":"crossref","unstructured":"Spitzbarth, I., Moore, S.A., Stein, V.M., Levine, J.M., Olby, N.J., Gjessing, K.M., Davidson, R.M., Lewis, M.J., Jeffery, N.D., and da Costa, R.C. (2020). Current Insights Into the Pathology of Canine Intervertebral Disc Extrusion-Induced Spinal Cord Injury. Front. Vet. Sci., 7.","DOI":"10.3389\/fvets.2020.595796"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"1857","DOI":"10.1111\/jvim.16179","article-title":"Serum C-Reactive Protein in Dogs with Paraplegia Secondary to Acute Intervertebral Disc Extrusion","volume":"35","author":"Foreman","year":"2021","journal-title":"J. Vet. Intern. Med."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"289","DOI":"10.1111\/j.1532-950X.2014.12144.x","article-title":"Evaluation of Serum Phosphorylated Neurofilament Subunit NF-H as a Prognostic Biomarker in Dogs with Thoracolumbar Intervertebral Disc Herniation","volume":"43","author":"Nishida","year":"2014","journal-title":"Vet. Surg."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"e1800047","DOI":"10.1002\/pmic.201800047","article-title":"Proteomics in Veterinary Medicine and Animal Science: Neglected Scientific Opportunities with Immediate Impact","volume":"18","author":"Galan","year":"2018","journal-title":"Proteomics"},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"4125118","DOI":"10.1155\/2024\/4125118","article-title":"Animal Wellness: The Power of Multiomics and Integrative Strategies","volume":"2024","author":"Ratan","year":"2024","journal-title":"Vet. Med. Int."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"1549","DOI":"10.1016\/j.spinee.2023.06.005","article-title":"Metabolomic Signature and Molecular Profile of Normal and Degenerated Human Intervertebral Disc Cells","volume":"23","author":"Francisco","year":"2023","journal-title":"Spine J."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"e70032","DOI":"10.1002\/jsp2.70032","article-title":"Analysis of Key Differential Metabolites in Intervertebral Disc Degeneration Based on Untargeted Metabolomics","volume":"8","author":"Zhou","year":"2025","journal-title":"JOR Spine"},{"key":"ref_44","doi-asserted-by":"crossref","unstructured":"Bo, T., and Fujii, J. (2025). Primary Roles of Branched Chain Amino Acids (BCAAs) and Their Metabolism in Physiology and Metabolic Disorders. Molecules, 30.","DOI":"10.3390\/molecules30010056"},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"7522","DOI":"10.1074\/jbc.TM117.000259","article-title":"Lipoic Acid Metabolism and Mitochondrial Redox Regulation","volume":"293","author":"Solmonson","year":"2018","journal-title":"J. Biol. Chem."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"702826","DOI":"10.3389\/fphys.2021.702826","article-title":"Branched-Chain Amino Acids: Catabolism in Skeletal Muscle and Implications for Muscle and Whole-Body Metabolism","volume":"12","author":"Mann","year":"2021","journal-title":"Front Physiol."},{"key":"ref_47","doi-asserted-by":"crossref","unstructured":"Zhang, Y., Guo, S., Xie, C., and Fang, J. (2020). Uridine Metabolism and Its Role in Glucose, Lipid, and Amino Acid Homeostasis. BioMed Res. Int., 2020.","DOI":"10.1155\/2020\/7091718"},{"key":"ref_48","doi-asserted-by":"crossref","unstructured":"Du, C., Liu, W.-J., Yang, J., Zhao, S.-S., and Liu, H.-X. (2022). The Role of Branched-Chain Amino Acids and Branched-Chain \u03b1-Keto Acid Dehydrogenase Kinase in Metabolic Disorders. Front. Nutr., 9.","DOI":"10.3389\/fnut.2022.932670"},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"111762","DOI":"10.1016\/j.nut.2022.111762","article-title":"Potential Roles of Branched-Chain Amino Acids in Neurodegeneration","volume":"103\u2013104","author":"Yoo","year":"2022","journal-title":"Nutrition"},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"1329","DOI":"10.1152\/ajpendo.00525.2011","article-title":"Leucine and MTORC1: A Complex Relationship","volume":"302","author":"Dodd","year":"2012","journal-title":"Am. J. Physiol. Endocrinol. Metab."},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"2145","DOI":"10.1007\/s00726-016-2240-1","article-title":"Leucine-Enriched Essential Amino Acids Attenuate Inflammation in Rat Muscle and Enhance Muscle Repair after Eccentric Contraction","volume":"48","author":"Kato","year":"2016","journal-title":"Amino Acids"},{"key":"ref_52","doi-asserted-by":"crossref","unstructured":"Parthasarathy, A., Cross, P.J., Dobson, R.C.J., Adams, L.E., Savka, M.A., and Hudson, A.O. (2018). A Three-Ring Circus: Metabolism of the Three Proteogenic Aromatic Amino Acids and Their Role in the Health of Plants and Animals. Front. Mol. Biosci., 5.","DOI":"10.3389\/fmolb.2018.00029"},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"207","DOI":"10.1016\/0026-0495(77)90057-9","article-title":"Effects of the Diet on Brain Neurotransmitters","volume":"26","author":"Fernstrom","year":"1977","journal-title":"Metabolism"},{"key":"ref_54","doi-asserted-by":"crossref","unstructured":"Li, D., Yu, S., Long, Y., Shi, A., Deng, J., Ma, Y., Wen, J., Li, X., Liu, S., and Zhang, Y. (2022). Tryptophan Metabolism: Mechanism-Oriented Therapy for Neurological and Psychiatric Disorders. Front. Immunol., 13.","DOI":"10.3389\/fimmu.2022.985378"},{"key":"ref_55","doi-asserted-by":"crossref","unstructured":"Yuan, Z., Zhao, X., Zhang, Y., Jiao, Y., Liu, Y., Gao, C., Zhang, J., Ma, Y., Wang, Z., and Li, T. (2025). Using Integrated Network Pharmacology and Metabolomics to Reveal the Mechanisms of the Combined Intervention of Ligustrazine and Sinomenine in CCI-Induced Neuropathic Pain Rats. Int. J. Mol. Sci., 26.","DOI":"10.3390\/ijms26062604"},{"key":"ref_56","doi-asserted-by":"crossref","first-page":"11140","DOI":"10.1073\/pnas.95.19.11140","article-title":"The Natural Polyamine Spermine Functions Directly as a Free Radical Scavenger","volume":"95","author":"Ha","year":"1998","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_57","doi-asserted-by":"crossref","unstructured":"Wang, L., Liu, Y., Qi, C., Shen, L., Wang, J., Liu, X., Zhang, N., Bing, T., and Shangguan, D. (2018). Oxidative Degradation of Polyamines by Serum Supplement Causes Cytotoxicity on Cultured Cells. Sci. Rep., 8.","DOI":"10.1038\/s41598-018-28648-8"},{"key":"ref_58","doi-asserted-by":"crossref","first-page":"104299","DOI":"10.1016\/j.phrs.2019.104299","article-title":"Spermine Oxidase: A Promising Therapeutic Target for Neurodegeneration in Diabetic Retinopathy","volume":"147","author":"Narayanan","year":"2019","journal-title":"Pharmacol. Res."},{"key":"ref_59","doi-asserted-by":"crossref","first-page":"30","DOI":"10.1016\/S0014-5793(01)02949-0","article-title":"Oxidation Products of Polyamines Induce Mitochondrial Uncoupling and Cytochrome c Release","volume":"507","author":"Maccarrone","year":"2001","journal-title":"FEBS Lett."},{"key":"ref_60","doi-asserted-by":"crossref","first-page":"2101","DOI":"10.1523\/JNEUROSCI.4351-10.2011","article-title":"Extracellular Spermine Exacerbates Ischemic Neuronal Injury through Sensitization of ASIC1a Channels to Extracellular Acidosis","volume":"31","author":"Duan","year":"2011","journal-title":"J. Neurosci."},{"key":"ref_61","doi-asserted-by":"crossref","first-page":"441","DOI":"10.1007\/s00726-011-1014-z","article-title":"Spermine Oxidase: Ten Years After","volume":"42","author":"Cervelli","year":"2012","journal-title":"Amino Acids"},{"key":"ref_62","doi-asserted-by":"crossref","unstructured":"Liu, F., Saul, A.B., Pichavaram, P., Xu, Z., Rudraraju, M., Somanath, P.R., Smith, S.B., Caldwell, R.B., and Narayanan, S.P. (2020). Pharmacological Inhibition of Spermine Oxidase Reduces Neurodegeneration and Improves Retinal Function in Diabetic Mice. J. Clin. Med., 9.","DOI":"10.3390\/jcm9020340"},{"key":"ref_63","doi-asserted-by":"crossref","first-page":"185","DOI":"10.1016\/j.biopha.2016.12.024","article-title":"Ornithine and Its Role in Metabolic Diseases: An Appraisal","volume":"86","author":"Sivashanmugam","year":"2017","journal-title":"Biomed. Pharmacother."},{"key":"ref_64","doi-asserted-by":"crossref","unstructured":"Li, T., Ning, N., Li, B., Luo, D., Qin, E., Yu, W., Wang, J., Yang, G., Nan, N., and He, Z. (2021). Longitudinal Metabolomics Reveals Ornithine Cycle Dysregulation Correlates With Inflammation and Coagulation in COVID-19 Severe Patients. Front. Microbiol., 12.","DOI":"10.3389\/fmicb.2021.723818"},{"key":"ref_65","first-page":"1145","article-title":"The Association Between Musculoskeletal Pain and Circulating Ornithine: A Population-Based Study","volume":"18","author":"Kautiainen","year":"2017","journal-title":"Pain Med."},{"key":"ref_66","doi-asserted-by":"crossref","first-page":"1911","DOI":"10.1007\/s00018-019-03363-3","article-title":"Proline-Dependent Regulation of Collagen Metabolism","volume":"77","author":"Karna","year":"2020","journal-title":"Cell. Mol. Life Sci."},{"key":"ref_67","doi-asserted-by":"crossref","unstructured":"Varma, S., Orgel, J.P.R.O., and Schieber, J.D. (2021). Contrasting Local and Macroscopic Effects of Collagen Hydroxylation. Int. J. Mol. Sci., 22.","DOI":"10.3390\/ijms22169068"},{"key":"ref_68","doi-asserted-by":"crossref","first-page":"10684","DOI":"10.1074\/jbc.M112.420042","article-title":"Medium-Chain Fatty Acid-Sensing Receptor, GPR84, Is a Proinflammatory Receptor","volume":"288","author":"Suzuki","year":"2013","journal-title":"J. Biol. Chem."},{"key":"ref_69","doi-asserted-by":"crossref","first-page":"47","DOI":"10.1007\/s10863-012-9481-9","article-title":"Long-Chain 3-Hydroxy Fatty Acids Accumulating in Long-Chain 3-Hydroxyacyl-CoA Dehydrogenase and Mitochondrial Trifunctional Protein Deficiencies Uncouple Oxidative Phosphorylation in Heart Mitochondria","volume":"45","author":"Tonin","year":"2013","journal-title":"J. Bioenerg. Biomembr."},{"key":"ref_70","doi-asserted-by":"crossref","unstructured":"Natarajan, S.K., and Ibdah, J.A. (2018). Role of 3-Hydroxy Fatty Acid-Induced Hepatic Lipotoxicity in Acute Fatty Liver of Pregnancy. Int. J. Mol. Sci., 19.","DOI":"10.3390\/ijms19010322"},{"key":"ref_71","doi-asserted-by":"crossref","unstructured":"Severyanova, L.A., Lazarenko, V.A., Plotnikov, D.V., Dolgintsev, M.E., and Kriukov, A.A. (2019). L-Lysine as the Molecule Influencing Selective Brain Activity in Pain-Induced Behavior of Rats. Int. J. Mol. Sci., 20.","DOI":"10.3390\/ijms20081899"},{"key":"ref_72","doi-asserted-by":"crossref","first-page":"904","DOI":"10.1016\/j.procbio.2016.04.009","article-title":"L-Lysine Regulates Tumor Necrosis Factor-Alpha and Matrix Metalloproteinase-3 Expression in Human Osteoarthritic Chondrocytes","volume":"51","author":"Huang","year":"2016","journal-title":"Process Biochem."},{"key":"ref_73","first-page":"637","article-title":"Medicinal Uses of L-Lysine: Past and Future","volume":"2","author":"Rao","year":"2011","journal-title":"Int. J. Res. Pharm. Sci."},{"key":"ref_74","doi-asserted-by":"crossref","first-page":"181","DOI":"10.1016\/S0887-8994(01)00407-6","article-title":"Pyridoxine-Dependent Seizures: Findings From Recent Studies Pose New Questions","volume":"26","author":"Gospe","year":"2002","journal-title":"Pediatr. Neurol."},{"key":"ref_75","doi-asserted-by":"crossref","first-page":"92","DOI":"10.1186\/s12974-018-1132-6","article-title":"Overexpression of Zinc-A2-Glycoprotein Suppressed Seizures and Seizure-Related Neuroflammation in Pentylenetetrazol-Kindled Rats","volume":"15","author":"Liu","year":"2018","journal-title":"J. Neuroinflammation"},{"key":"ref_76","doi-asserted-by":"crossref","unstructured":"Mart\u00ednez-Navarro, I., Vilchis-Gil, J., Coss\u00edo-Torres, P.E., Hern\u00e1ndez-Mendoza, H., Kl\u00fcnder-Kl\u00fcnder, M., Layseca-Espinosa, E., Galicia-Cruz, O.G., and Rios-Lugo, M.J. (2024). Serum Zinc-Alpha-2 Glycoprotein and Zinc Levels and Their Relationship with Insulin Resistance and Biochemical Parameters in Overweight and Obese Children. Biol. Trace Elem. Res.","DOI":"10.1007\/s12011-024-04480-2"},{"key":"ref_77","first-page":"1","article-title":"Relationship between Neuropathic Pain and Zinc Ion","volume":"1","author":"Kitayama","year":"2016","journal-title":"Glob. Drugs Ther."},{"key":"ref_78","doi-asserted-by":"crossref","first-page":"117351","DOI":"10.1016\/j.lfs.2020.117351","article-title":"Zinc Promotes Functional Recovery after Spinal Cord Injury by Activating Nrf2\/HO-1 Defense Pathway and Inhibiting Inflammation of NLRP3 in Nerve Cells","volume":"245","author":"Li","year":"2020","journal-title":"Life Sci."},{"key":"ref_79","doi-asserted-by":"crossref","first-page":"3356","DOI":"10.1007\/s12011-022-03441-x","article-title":"Zinc Concentration in Blood Serum of Healthy Dogs","volume":"201","year":"2023","journal-title":"Biol. Trace Elem. Res."},{"key":"ref_80","doi-asserted-by":"crossref","first-page":"1600082","DOI":"10.1002\/prca.201600082","article-title":"Elevated Levels of Protein AMBP in Cerebrospinal Fluid of Women with Preeclampsia Compared to Normotensive Pregnant Women","volume":"11","author":"Duvekot","year":"2017","journal-title":"Proteom. Clin. Appl."},{"key":"ref_81","doi-asserted-by":"crossref","unstructured":"Ruzha, Y., Ni, J., Quan, Z., Li, H., and Qing, H. (2022). Role of Vitronectin and Its Receptors in Neuronal Function and Neurodegenerative Diseases. Int. J. Mol. Sci., 23.","DOI":"10.3390\/ijms232012387"},{"key":"ref_82","doi-asserted-by":"crossref","first-page":"202580","DOI":"10.1242\/jcs.202580","article-title":"Blood Vitronectin Is a Major Activator of LIF and IL-6 in the Brain through Integrin-FAK and UPAR Signaling","volume":"131","author":"Keasey","year":"2018","journal-title":"J. Cell Sci."},{"key":"ref_83","doi-asserted-by":"crossref","first-page":"e1289","DOI":"10.1002\/jsp2.1289","article-title":"Understanding the Etiopathogenesis of Lumbar Intervertebral Disc Herniation: From Clinical Evidence to Basic Scientific Research","volume":"7","author":"Zhou","year":"2024","journal-title":"JOR Spine"},{"key":"ref_84","doi-asserted-by":"crossref","first-page":"30","DOI":"10.1038\/s41572-021-00264-3","article-title":"Trauma-Induced Coagulopathy","volume":"7","author":"Moore","year":"2021","journal-title":"Nat. Rev. Dis. Primers"},{"key":"ref_85","doi-asserted-by":"crossref","first-page":"2017","DOI":"10.1111\/j.1538-7836.2008.03181.x","article-title":"The Multi-Functional Serpin, Protein C Inhibitor: Beyond Thrombosis and Hemostasis","volume":"6","author":"Suzuki","year":"2008","journal-title":"J. Thromb. Haemost."},{"key":"ref_86","doi-asserted-by":"crossref","first-page":"324","DOI":"10.1016\/0268-9499(93)90148-O","article-title":"Fibrinolytic Changes During Acute Vascular Damage Induced by Mediterranean Spotted Fever","volume":"7","author":"Vicente","year":"1993","journal-title":"Fibrinolysis"},{"key":"ref_87","doi-asserted-by":"crossref","first-page":"558","DOI":"10.1182\/blood.2019002224","article-title":"Protease Activity in Single-Chain Prekallikrein","volume":"135","author":"Ivanov","year":"2020","journal-title":"Blood"},{"key":"ref_88","doi-asserted-by":"crossref","first-page":"5302","DOI":"10.1182\/blood-2011-05-355248","article-title":"Selective Depletion of Plasma Prekallikrein or Coagulation Factor XII Inhibits Thrombosis in Mice without Increased Risk of Bleeding","volume":"118","author":"Revenko","year":"2011","journal-title":"Blood"},{"key":"ref_89","doi-asserted-by":"crossref","unstructured":"Jaffa, A.A., Jaffa, M.A., Moussa, M., Ahmed, I.A., Karam, M., Aldeen, K.S., Al Sayegh, R., El-Achkar, G.A., Nasrallah, L., and Yehya, Y. (2021). Modulation of Neuro-Inflammatory Signals in Microglia by Plasma Prekallikrein and Neuronal Cell Debris. Front. Pharmacol., 12.","DOI":"10.3389\/fphar.2021.743059"},{"key":"ref_90","doi-asserted-by":"crossref","first-page":"353","DOI":"10.1007\/s00726-009-0431-8","article-title":"Potential Anticancer Application of Polyamine Oxidation Products Formed by Amine Oxidase: A New Therapeutic Approach","volume":"38","author":"Agostinelli","year":"2010","journal-title":"Amino Acids"},{"key":"ref_91","doi-asserted-by":"crossref","first-page":"2477","DOI":"10.1096\/fj.201600935R","article-title":"Amine Oxidase Activity Regulates the Development of Pulmonary Fibrosis","volume":"31","author":"Elima","year":"2017","journal-title":"FASEB J."},{"key":"ref_92","doi-asserted-by":"crossref","first-page":"669","DOI":"10.14245\/ns.2244666.333","article-title":"The Role of GABA in Spinal Cord Injury","volume":"19","author":"Yao","year":"2022","journal-title":"Neurospine"}],"container-title":["Metabolites"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2218-1989\/15\/6\/396\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,9]],"date-time":"2025-10-09T17:50:55Z","timestamp":1760032255000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2218-1989\/15\/6\/396"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,6,12]]},"references-count":92,"journal-issue":{"issue":"6","published-online":{"date-parts":[[2025,6]]}},"alternative-id":["metabo15060396"],"URL":"https:\/\/doi.org\/10.3390\/metabo15060396","relation":{},"ISSN":["2218-1989"],"issn-type":[{"type":"electronic","value":"2218-1989"}],"subject":[],"published":{"date-parts":[[2025,6,12]]}}}