Objective: To investigate the metabolic changes after methotrexate (MTX) exposure in mice using a gas chromatography-mass spectrometry (GC-MS)-based untargeted metabolomics approach. Methods: MTX (20 mg·kg-1) was intraperitoneally injected into experimental mice (n=9) at the 7th day, and mice in control group (n=9) were treated with the same amount of saline. A gas chromatography-mass spectrometry (GC-MS) approach was employed to identify discriminant metabolites in serum and various organs including the heart, liver, kidney, lung, intestine, stomach, and hippocampus. The potential metabolites were identified using orthogonal partial least squares discrimination analysis (OPLS-DA). Subsequently, the MetaboAnalyst 5.0 (http://www.metaboanalyst.ca) and Kyoto Encyclopedia of genes and genomes database (KEGG, http://www.kegg.jp) were employed to depict the metabolic pathways. Pathological analysis and metabolite verification were carried our using intestinal tissue as an example. Results: Untargeted metabolomics analysis showed that MTX exposure affected the comprehensive metabolic profiling of mice, especially in the liver and intestine. Pathway analysis identified that MTX affected several metabolic pathways in mice, such as amino acid metabolism/ synthesis, energy metabolism, pantothenate and coenzyme A biosynthesis, pyrimidine metabolism, and glutathione metabolism. MTX treatment can obviously damage the intestinal epithelial structure and increase the numbers of inflammatory cells. Meanwhile, it was found that the contents of L-glutamic acid, L-aspartic acid and glycine were significantly increased after MTX treatment, which was consistent with the results of metabonomic analysis. Conclusion: This study clarified the metabolic profiling of methotrexate exposure on various tissues of mouse, identified metabolic biomarkers and metabolic pathways related to MTX toxicity, and provided new insights for the MTX toxicological mechanism.
MIN Tuo, DU Zhong-yan, ZHAO Yan-xia
. GC-MS-based untargeted metabolomics approach for metabolic profiling of methotrexate-induced toxicity*[J]. Chinese Journal of Pharmaceutical Analysis, 2023
, 43(9)
: 1553
-1564
.
DOI: 10.16155/j.0254-1793.2023.09.13
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