Summary of Study ST000379

This data is available at the NIH Common Fund's National Metabolomics Data Repository (NMDR) website, the Metabolomics Workbench, https://www.metabolomicsworkbench.org, where it has been assigned Project ID PR000298. The data can be accessed directly via it's Project DOI: 10.21228/M8HW28 This work is supported by NIH grant, U2C- DK119886.

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Study IDST000379
Study TitleTemporal metabolomic responses of cultured HepG2 liver cells to high fructose and high glucose exposures
Study SummaryHigh fructose consumption has been implicated with deleterious effects on human health, including hyperlipidemia elicited through de novo lipogenesis. However, more global effects of fructose on cellular metabolism have not been elucidated. In order to explore the metabolic impact of fructose-containing nutrients, we applied both GC-TOF and HILIC-QTOF mass spectrometry metabolomic strategies using extracts from cultured HepG2 cells exposed to fructose, glucose, or fructose + glucose. Cellular responses were analyzed in a time-dependent manner, incubated in media containing 5.5 mM glucose + 5.0 mM fructose in comparison to controls incubated in media containing either 5.5 mM glucose or 10.5 mM glucose. Mass spectrometry identified 156 unique known metabolites and a large number of unknown compounds, which revealed metabolite changes due to both utilization of fructose and high-carbohydrate loads independent of hexose structure. Fructose was shown to be partially converted to sorbitol, and generated higher levels of fructose-1-phosphate as a precursor for glycolytic intermediates. Differentially regulated ratios of 3-phosphoglycerate to serine pathway intermediates in high fructose media indicated a diversion of carbon backbones away from energy metabolism. Additionally, high fructoseconditions changed levels of complex lipids toward phosphatidylethanolamines. Patterns of acylcarnitines in response to high hexose exposure (10.5 mM glucose or glucose/fructose combination) suggested a reduction in mitochondrial beta-oxidation.
Institute
University of California, Davis
DepartmentGenome and Biomedical Sciences Facility
LaboratoryWCMC Metabolomics Core
Last NameFiehn
First NameOliver
Address1315 Genome and Biomedical Sciences Facility, 451 Health Sciences Drive, Davis, CA 95616
Emailofiehn@ucdavis.edu
Phone(530) 754-8258
Submit Date2016-04-14
Raw Data AvailableYes
Raw Data File Type(s)bin
Analysis Type DetailGC-MS
Release Date2016-04-25
Release Version1
Oliver Fiehn Oliver Fiehn
https://dx.doi.org/10.21228/M8HW28
ftp://www.metabolomicsworkbench.org/Studies/ application/zip

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Sample Preparation:

Sampleprep ID:SP000407
Sampleprep Summary:Sample material was thawed on ice, vortexed for 20 s, sonicated for 5 min with a VWR 50HT Ultrasonic Bath (VWR International Inc., Bridgeport, NJ), and separated into 500 μL aliquots. Each aliquot was centrifuged for 5 min @ 14,000 rcf, and supernatant was collected and lyophilized to dryness. Samples was kept on ice and removed only for sonication, centrifugation, and lyophilization steps. Lyophilized material was used for HILIC-QTOF metabolite profiling without additional clean-up steps. Lyophilized material for GC-TOF analysis was redissolved in 1:1 acetonitrile/H2O, vortexed for 10 s, and centrifuged for 5 min @ 14,000 rcf. Supernatant was collected and lyophilized to dryness.
Sampleprep Protocol Filename:metabolomic_responses_of_cultured_HepG2_liver_cells.pdf
Processing Method:Lysophilization
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