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Metabolic Characterization of Patients with NASH and ASH by High-Resolution Multinuclear NMR Spectroscopy on Bodyfluids


Zwingmann C, Gottschalk,  S
Department High-Field Magnetic Resonance, Max Planck Institute for Biological Cybernetics, Max Planck Society;

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Hohnholt, M., Zwingmann C, Gottschalk, S., Boulanger, Y., & Bilodeau, M. (2007). Metabolic Characterization of Patients with NASH and ASH by High-Resolution Multinuclear NMR Spectroscopy on Bodyfluids. Talk presented at Canadian Digestive Diseases Week: 45th Annual Meeting of the Canadian Association of Gastroenterology (CDDW 2007). Banff, Canada.

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INTRODUCTION: Liver steatosis is a common cause of liver disease. Non-alcoholic fatty liver disease (NASH or NAFLD) ranges from steatosis without inflammation to steatohepatitis, ballooning degeneration with or without liver fibrosis. Similar pathological findings are observed in patients with ASH (alcoholic steatohepatitis), and it is difficult to differentiate both pathologies by clinical and biochemical evaluations. High-resolution 1H-NMR has emerged as a powerful technique to simultaneously identify and quantify multiple metabolites of medical significance without a requirement for pre-selection or separation of metabolites. AIM: Since it is becoming urgent to develop new diagnostic methods for NAFLD, we applied novel multinuclear NMR methods to detect the metabolic profile in body fluids in patients with NASH and ASH. METHODS: Thirty non-alcoholic patients, 7 alcoholic steatotic patients and 30 age-matched control subjects were recruited. Urine samples were lyophilized. We used dual-extraction methods to investigate both water-soluble metabolites and lipophilic compounds involved in fatty acid- and lipid metabolism, in blood plasma and serum. Furthermore, to study metabolites not accessible by conventional 1H NMR analysis, natural abundance 13C NMR measurements were performed. To identify unknown metabolites in body fluids, two-dimensional 1H-1H} and {1H-13C experiments were used. RESULTS: 1H- and 13C-NMR spectra of urine and blood extracts clearly showed considerable differences in specific metabolites involved in liver intermediary metabolism in patients with NASH and ASH compared to healthy controls. Selective changes in patients with ASH compared to controls were detected urine (decreases of hippurate (to 46±13), and urea (to 37±21), increases of TMAO (to 157±28) and tyrosine (to 322±124) as well as accumulation of bile acids) and in blood plasma (increases of TMAO (to 314±58) and methionine (to 620±81) and decreases in branched-chain amino acids (to 32±5)). In patients with NASH, 13C-NMR spectra showed several and significant changes in metabolites involved in mitochondrial and lipid metabolism. CONCLUSIONS: New applications of NMR methods on human body fluids are of great potential to characterize NAFLD, a disease displaying multiple interrelated metabolic factors. This approach could provide new data to characterize steatosis, help to distinguish between NASH and ASH, and also give insights in the pathophysiology of both diseases.