Global Metabolomic and IsobaricTagging CapillaryLiquid Chromatography–Tandem Mass Spectrometry Approaches forUncovering Pathway Dysfunction in Diabetic Mouse Aorta

摘要

Despite the prevalence of diabetes and the global health risks it poses, the biochemical pathogenesis of diabetic complications remains poorly understood with few effective therapies. This study employs capillary liquid chromatography (capLC) and tandem mass spectrometry (MS/MS) in conjunction with both global metabolomics and isobaric tags specific to amines and carbonyls to probe aortic metabolic content in diabetic mice with hyperglycemia, hyperlipidemia, hypertension, and stenotic vascular damage. Using these combined techniques, metabolites well-characterized in diabetes as well as novel pathways were investigated. A total of 53 986 features were detected, 719 compounds were identified as having significant fold changes (thresholds ≥2 or ≤0.5), and 48 metabolic pathways were found to be altered with at least 2 metabolite hits in diabetic samples. Pathways related to carbonyl stress, carbohydrate metabolism, and amino acid metabolism showed the greatest number of metabolite changes. Three novel pathways with previously limited or undescribed roles in diabetic complications—vitaminB6, propanoate, and butanoate metabolism—were also shown tobe altered in multiple points along the pathway. These discoveriessupport the theory that diabetic vascular complications arise fromthe interplay of a myriad of metabolic pathways in conjunction withoxidative and carbonyl stress, which may provide not only new andmuch needed biomarkers but also insights into novel therapeutic targets.