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.
See: https://www.metabolomicsworkbench.org/about/howtocite.php
| Study ID | ST000379 |
| Study Title | Temporal metabolomic responses of cultured HepG2 liver cells to high fructose and high glucose exposures |
| Study Summary | High 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 |
| Department | Genome and Biomedical Sciences Facility |
| Laboratory | WCMC Metabolomics Core |
| Last Name | Fiehn |
| First Name | Oliver |
| Address | 1315 Genome and Biomedical Sciences Facility, 451 Health Sciences Drive, Davis, CA 95616 |
| ofiehn@ucdavis.edu | |
| Phone | (530) 754-8258 |
| Submit Date | 2016-04-14 |
| Raw Data Available | Yes |
| Raw Data File Type(s) | bin |
| Analysis Type Detail | GC-MS |
| Release Date | 2016-04-25 |
| Release Version | 1 |
Select appropriate tab below to view additional metadata details:
Project:
| Project ID: | PR000298 |
| Project DOI: | doi: 10.21228/M8HW28 |
| Project Title: | Temporal metabolomic responses of cultured HepG2 liver cells to high fructose and high glucose exposures |
| Project Type: | Time-course Study |
| Project Summary: | High 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 |
| Department: | Genome and Biomedical Sciences Facility |
| Laboratory: | WCMC Metabolomics Core |
| Last Name: | Fiehn |
| First Name: | Oliver |
| Address: | 1315 Genome and Biomedical Sciences Facility, 451 Health Sciences Drive, Davis, CA 95616 |
| Email: | ofiehn@ucdavis.edu |
| Phone: | (530) 754-8258 |
| Funding Source: | NIH U24DK097154 |
Subject:
| Subject ID: | SU000400 |
| Subject Type: | Cells |
| Subject Species: | Homo sapiens |
| Taxonomy ID: | 9606 |
| Species Group: | Human |
Factors:
Subject type: Cells; Subject species: Homo sapiens (Factor headings shown in green)
| mb_sample_id | local_sample_id | Treatment | Time |
|---|---|---|---|
| SA017079 | 110625baasa27_1 | 10.5mM Glu | 10min |
| SA017080 | 110625baasa04_1 | 10.5mM Glu | 10min |
| SA017081 | 110624baasa30_1 | 10.5mM Glu | 10min |
| SA017082 | 110624baasa23_1 | 10.5mM Glu | 10min |
| SA017083 | 110625baasa14_1 | 10.5mM Glu | 10min |
| SA017084 | 110625baasa11_1 | 10.5mM Glu | 10min |
| SA017085 | 110624baasa12_1 | 10.5mM Glu | 1hr |
| SA017086 | 110625baasa03_1 | 10.5mM Glu | 1hr |
| SA017087 | 110625baasa19_1 | 10.5mM Glu | 1hr |
| SA017088 | 110624baasa46_1 | 10.5mM Glu | 1hr |
| SA017089 | 110624baasa15_1 | 10.5mM Glu | 1hr |
| SA017090 | 110625baasa22_1 | 10.5mM Glu | 1hr |
| SA017091 | 110624baasa33_1 | 10.5mM Glu | 24hr |
| SA017092 | 110624baasa35_1 | 10.5mM Glu | 24hr |
| SA017093 | 110624baasa24_1 | 10.5mM Glu | 24hr |
| SA017094 | 110625baasa18_1 | 10.5mM Glu | 24hr |
| SA017095 | 110624baasa47_1 | 10.5mM Glu | 24hr |
| SA017096 | 110624baasa03_1 | 10.5mM Glu | 24hr |
| SA017097 | 110624baasa20_1 | 10.5mM Glu | 6hr |
| SA017098 | 110624baasa05_1 | 10.5mM Glu | 6hr |
| SA017099 | 110624baasa42_1 | 10.5mM Glu | 6hr |
| SA017100 | 110624baasa31_1 | 10.5mM Glu | 6hr |
| SA017101 | 110625baasa21_1 | 10.5mM Glu | 6hr |
| SA017102 | 110625baasa09_1 | 10.5mM Glu | 6hr |
| SA017103 | 110624baasa44_1 | 5.5mM Glu 5mM Fru | 10min |
| SA017104 | 110624baasa16_1 | 5.5mM Glu 5mM Fru | 10min |
| SA017105 | 110624baasa27_1 | 5.5mM Glu 5mM Fru | 10min |
| SA017106 | 110624baasa43_1 | 5.5mM Glu 5mM Fru | 10min |
| SA017107 | 110625baasa01_1 | 5.5mM Glu 5mM Fru | 10min |
| SA017108 | 110624baasa28_1 | 5.5mM Glu 5mM Fru | 10min |
| SA017109 | 110625baasa23_1 | 5.5mM Glu 5mM Fru | 1hr |
| SA017110 | 110624baasa01_1 | 5.5mM Glu 5mM Fru | 1hr |
| SA017111 | 110625baasa08_1 | 5.5mM Glu 5mM Fru | 1hr |
| SA017112 | 110624baasa08_1 | 5.5mM Glu 5mM Fru | 1hr |
| SA017113 | 110625baasa15_1 | 5.5mM Glu 5mM Fru | 1hr |
| SA017114 | 110624baasa10_1 | 5.5mM Glu 5mM Fru | 1hr |
| SA017115 | 110624baasa40_1 | 5.5mM Glu 5mM Fru | 24hr |
| SA017116 | 110624baasa06_1 | 5.5mM Glu 5mM Fru | 24hr |
| SA017117 | 110624baasa37_1 | 5.5mM Glu 5mM Fru | 24hr |
| SA017118 | 110625baasa28_1 | 5.5mM Glu 5mM Fru | 24hr |
| SA017119 | 110625baasa07_1 | 5.5mM Glu 5mM Fru | 24hr |
| SA017120 | 110624baasa45_1 | 5.5mM Glu 5mM Fru | 24hr |
| SA017121 | 110625baasa05_1 | 5.5mM Glu 5mM Fru | 6hr |
| SA017122 | 110624baasa19_1 | 5.5mM Glu 5mM Fru | 6hr |
| SA017123 | 110624baasa39_1 | 5.5mM Glu 5mM Fru | 6hr |
| SA017124 | 110624baasa17_1 | 5.5mM Glu 5mM Fru | 6hr |
| SA017125 | 110624baasa13_1 | 5.5mM Glu 5mM Fru | 6hr |
| SA017126 | 110624baasa49_1 | 5.5mM Glu 5mM Fru | 6hr |
| SA017127 | 110624baasa34_1 | 5.5mM Glu | 10min |
| SA017128 | 110624baasa09_1 | 5.5mM Glu | 10min |
| SA017129 | 110625baasa25_1 | 5.5mM Glu | 10min |
| SA017130 | 110624baasa14_1 | 5.5mM Glu | 10min |
| SA017131 | 110625baasa16_1 | 5.5mM Glu | 10min |
| SA017132 | 110624baasa50_1 | 5.5mM Glu | 10min |
| SA017133 | 110624baasa02_1 | 5.5mM Glu | 1hr |
| SA017134 | 110625baasa17_1 | 5.5mM Glu | 1hr |
| SA017135 | 110624baasa41_1 | 5.5mM Glu | 1hr |
| SA017136 | 110624baasa11_1 | 5.5mM Glu | 1hr |
| SA017137 | 110624baasa26_1 | 5.5mM Glu | 1hr |
| SA017138 | 110624baasa22_1 | 5.5mM Glu | 1hr |
| SA017139 | 110624baasa38_1 | 5.5mM Glu | 24hr |
| SA017140 | 110625baasa20_1 | 5.5mM Glu | 24hr |
| SA017141 | 110625baasa24_1 | 5.5mM Glu | 24hr |
| SA017142 | 110625baasa02_1 | 5.5mM Glu | 24hr |
| SA017143 | 110625baasa10_1 | 5.5mM Glu | 24hr |
| SA017144 | 110624baasa48_1 | 5.5mM Glu | 24hr |
| SA017145 | 110624baasa18_1 | 5.5mM Glu | 6hr |
| SA017146 | 110624baasa21_1 | 5.5mM Glu | 6hr |
| SA017147 | 110624baasa04_1 | 5.5mM Glu | 6hr |
| SA017148 | 110624baasa25_1 | 5.5mM Glu | 6hr |
| SA017149 | 110625baasa06_1 | 5.5mM Glu | 6hr |
| SA017150 | 110625baasa13_1 | 5.5mM Glu | 6hr |
| SA017151 | 110624baasa36_1 | Glucosamine | 10min |
| SA017152 | 110624baasa32_1 | Glucosamine | 10min |
| SA017153 | 110624baasa29_1 | Glucosamine | 10min |
| SA017154 | 110625baasa26_1 | Glucosamine | 10min |
| SA017155 | 110624baasa07_1 | Glucosamine | 10min |
| SA017156 | 110625baasa12_1 | Glucosamine | 10min |
| Showing results 1 to 78 of 78 |
Collection:
| Collection ID: | CO000394 |
| Collection Summary: | HepG2 cells (ATCC HB-8065) were cultured in MEM containing 10 % (v:v) FBS, 100 U/mL penicillin and 100 mg/mL streptomycin (Invitrogen, Carlsbad, CA), 1 x MEM non-essential amino acids, and 5.5 mM glucose at 37 °C in a 5 % CO2 environment. Cells were grown for four to six passages in 10 cm tissue culture dishes with 14 mL MEM, and transferred to MULTIWELL™ 12 well culture dishes for incubation in 2 mL of the treatment medium. Upon reaching 80 % confluency, the cell culture medium was changed to media representative of experimental condition: 5.5 mM glucose MEM, 5.5 mM glucose MEM + 5 mM glucose, or 5.5 mM glucose MEM + 5 mM fructose. Media was replenished after 24 and 48 h. Cell material was collected at 10 min, 1, 6, and 24 h time points following the 48 h media replacement. Culture dishes were placed on ice and each well was washed twice with 1 mL ice-cold PBS. 4 mL ice-cold 3:1 methanol/H2O extraction solvent was added to each well. Cell material was manually scraped from each well, and the extraction solvent cell material suspension was transferred to collection tubes and frozen at −80 °C prior to further processing. |
| Collection Protocol Filename: | metabolomic_responses_of_cultured_HepG2_liver_cells.pdf |
| Sample Type: | Cell |
| Collection Location: | Invitrogen, Carlsbad, CA |
| Collection Frequency: | Cell material was collected at 10 min, 1, 6, and 24 h time points following the 48 h media replacement. |
| Tissue Cell Identification: | HepG2 |
Treatment:
| Treatment ID: | TR000414 |
| Treatment Summary: | 3 Treatments: A) Control group of HepG2 cells incubated in media with 5.5 mM glucose (Glc5) B) HepG2 cells incubated in media containing 5.5 mM glucose + 5.0 mM fructose (Glc5Fru5) C) HepG2 cells incubated in media containing 10.5 mM glucose (Glc10) |
| Treatment Protocol Filename: | metabolomic_responses_of_cultured_HepG2_liver_cells.pdf |
| Treatment Protocol Comments: | HepG2 cells incubated in media containing 10.5 mM glucose (Glc10) was included to enable differentiation of metabolic effects caused by fructose from metabolic effects caused by increased hexose resources |
| Cell Media: | Cells were acclimated to their respective media condition for 48 h prior to collection of sample material to obtain a metabolite profile representative of continued exposure instead of response to a sudden change in carbohydrate resources. Media was replenished after 24 and 48 h |
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 |
Combined analysis:
| Analysis ID | AN000613 |
|---|---|
| Analysis type | MS |
| Chromatography type | GC |
| Chromatography system | Agilent 6890N |
| Column | Restek Corporation Rtx-5Sil MS |
| MS Type | EI |
| MS instrument type | GC-TOF |
| MS instrument name | Leco Pegasus IV TOF |
| Ion Mode | POSITIVE |
| Units | counts |
Chromatography:
| Chromatography ID: | CH000439 |
| Methods Filename: | Data_Dictionary_Fiehn_laboratory_GCTOF_MS_primary_metabolism_10-15-2013_general.pdf |
| Instrument Name: | Agilent 6890N |
| Column Name: | Restek Corporation Rtx-5Sil MS |
| Column Temperature: | 50-330C |
| Flow Rate: | 1 ml/min |
| Oven Temperature: | 50°C for 1 min, then ramped at 20°C/min to 330°C, held constant for 5 min |
| Transferline Temperature: | 230C |
| Washing Buffer: | Ethyl Acetate |
| Sample Loop Size: | 30 m length x 0.25 mm internal diameter |
| Randomization Order: | Excel generated |
| Chromatography Type: | GC |
MS:
| MS ID: | MS000546 |
| Analysis ID: | AN000613 |
| Instrument Name: | Leco Pegasus IV TOF |
| Instrument Type: | GC-TOF |
| MS Type: | EI |
| Ion Mode: | POSITIVE |
| Ion Source Temperature: | 250 C |
| Ionization Energy: | 70 eV |
| Mass Accuracy: | Nominal |
| Source Temperature: | 250 C |
| Scan Range Moverz: | 85-500 Da |
| Scanning Cycle: | 17 Hz |
| Scanning Range: | 85-500 Da |
