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A method for the simultaneous determination of 3T3-L1 adipocyte metabolites by liquid chromatography/mass spectrometry using [(13)C]-stable isotopes
Authors:Nagai Hiroyuki  Goto Tsuyoshi  Takahashi Nobuyuki  Kusudoh Tatsuya  Deyashiki Yoshihiro  Esaka Yukihiro  Kawada Teruo
Institution:Laboratory of Molecular Function of Food, Division of Food Science and Biotechnology, Graduate School of Agriculture, Kyoto University, Uji, Kyoto, Japan. managai@blue.ocn.ne.jp
Abstract:A useful method employing liquid chromatography mass spectrometry (LC/MS) and a stable isotope was developed for simultaneous examination of major metabolism in adipocytes, de novo fatty acid synthesis, glycerol output, and glucose uptake with high sensitivity. The addition of thiazolidinediones, potent agonists of peroxisome proliferator-activated receptors-γ, for 10 d increased glucose uptake in a dose-dependent manner. Fatty acid (FA) synthesis increased at low concentrations of thiazolidinediones (TZDs) and decreased at high concentrations. It is important to assess adipocytes from various examples of metabolism, because each example of adipocyte metabolism is directly related to obesity or metabolic syndrome in various ways. The technique makes metabolic examination easier than conventional methods by means of radioisotopes and makes it possible to identify metabolites and to apply them in biomarker screening.
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