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Purpose: Two-dimensional mapping of arterial tissue by MALDI-MSI for in-situ visualization of proteins, metabolites and lipids.Methods: Human pathological arteries and healthy mammary arteries were obtained from patients subjected to carotid endarterectomy and from patients admitted for coronary artery bypass surgery, respectively. An early model of atherosclerosis was performed by feeding rabbits with regular diet (control group) and with cholesterol-rich diet (pathological group). Following histological characterization, specific methods for visualization of metabolites, proteins and lipids in their original locations inside the different arterial layers (intima, media) were set-up. Aortic tissue from both animal groups was dissected and comparatively analyzed by MALDI-MSI, using sinapinic acid, 2,5-dihidroxybenzoic acid and 9-aminoacridine as matrix for proteins, lipids and metabolites respectively. Experiments were performed in an UltrafleXtreme MALDI-ToF/ToF and a Solarix FT-ICR (BrukerDaltonik) instruments.Results: Protein evaluation resulted in 15 m/z values defining media or intima layers. From them six are significant altered between control and pathological animals. Two of them are localized in the media layer and the other four in the intima. There are 8 m/z values of metabolites identifying lipids. All of them are localized in the intima layer and are significantly altered when comparing control and atherosclerotic tissue. Lipids analysis show 11 masses (m/z) defining intima, 8 of them specifically localized in the plaque region. A couple of them are varied with significance.Conclusions: A novel protein MALDI-MSI protocol for human atherosclerotic lesions and healthy arteries has been developed and successfully applied. A panel of 15 m/z consisting in proteins, metabolites and lipids are here described to be significant involved in atherosclerosis and, importantly, its identification goes together with their localization, helping enormously in the study of underlying mechanism of atherosclerosis. [ABSTRACT FROM AUTHOR]

Purpose: Search of progression and recovery markers in atherosclerosis disease.Methods: An animal model of early atherosclerosis was done in rabbits fed with different diets, regular diet and cholesterol-rich (control and pathological group).Urine samples from animals were analyzed in the search for alterations of the proteome and metabolome (by 2DE-DIGE and NMR, respectively). Confirmation analyses were performed by SRM-(LC-MS/MS).In a pilot translational study, human urine samples from healthy donors and NSTEACS patients at the onset and discharge were analyzed.Results: Four proteins were found significant altered between the two animal groups. Cathepsin D is increased in urine from pathological animals and Hemopexin, Kallikrein 1 (KLK1) and Zymogen granule protein 16B (ZG16B) are decreased. After translational studies KLK1 and ZG16B are proposed to be markers of both, progression and recovery. NMR pops up twenty metabolites significantly varied at early stages of atherosclerosis between animal groups. Eight metabolites increased in urine from pathological animals and twelve metabolites showed the opposite trend (decreased). Two panels of six and three metabolites are found as responders to atherosclerosis progression or recovery from an acute event, respectively. Metabolism changes of particular sugars, hydroxy acids, amino acids, cyclic alcohols, polyamines and imidazolidines are here shown to be associated to atherosclerosis.Conclusions: These data confirm that omics approaches are extremely valuable for the diagnosis and monitoring of atherosclerotic pathologies. Four proteins and twenty metabolites are significantly altered in an early animal model of atherosclerosis. In human samples, two novel urinary panels of proteins and metabolites are here first shown able to monitor atherosclerosis progression and recovery from an acute event. [ABSTRACT FROM PUBLISHER]