Summary of Study ST000656

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 PR000461. The data can be accessed directly via it's Project DOI: 10.21228/M8G609 This work is supported by NIH grant, U2C- DK119886.

See: https://www.metabolomicsworkbench.org/about/howtocite.php

This study contains a large results data set and is not available in the mwTab file. It is only available for download via FTP as data file(s) here.

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Study IDST000656
Study TitleOmega-6 and omega-3 oxylipins are implicated in soybean oil-induced obesity in mice (part III)
Study SummaryIn this study we have compared the metabolic effects of conventional soybean oil to those of genetically modified Plenish soybean oil, that is low in linoleic acid and high in oleic acid. This work builds on our previous study showing that soybean oil, rich in polyunsaturated fats, is more obesogenic and diabetogenic than coconut oil, rich in saturated fats (PMID: 26200659). Here, in order to elucidate the mechanisms responsible for soybean oil induced obesity, we have performed the first ever metabolomics (in plasma and liver) and proteomics on the livers of mice fed the two soybean oil diets (plus those fed a high coconut oil and Viv chow diet). Our results show that the new high oleic soybean oil induces less obesity and adiposity than conventional soybean oil, but can cause hepatomegaly and liver dysfunction. Metabolomic analysis reveals that the hepatic and plasma metabolic profiles differ considerably between the two soybean oils. Hepatic C18 oxylipin metabolites of omega-6 (ω6) and omega-3 (ω3) fatty acids (linoleic and α-linolenic acid, respectively) in the cytochrome P450/soluble epoxide hydrolase pathway were found to correlate positively with obesity.
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 Date2017-06-23
Analysis Type DetailLC-MS
Release Date2017-07-01
Release Version1
Oliver Fiehn Oliver Fiehn
https://dx.doi.org/10.21228/M8G609
ftp://www.metabolomicsworkbench.org/Studies/ application/zip

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Project:

Project ID:PR000461
Project DOI:doi: 10.21228/M8G609
Project Title:Omega-6 and omega-3 oxylipins are implicated in soybean oil-induced obesity in mice
Project Summary:In this study we have compared the metabolic effects of conventional soybean oil to those of genetically modified Plenish soybean oil, that is low in linoleic acid and high in oleic acid. This work builds on our previous study showing that soybean oil, rich in polyunsaturated fats, is more obesogenic and diabetogenic than coconut oil, rich in saturated fats (PMID: 26200659). Here, in order to elucidate the mechanisms responsible for soybean oil induced obesity, we have performed the first ever metabolomics (in plasma and liver) and proteomics on the livers of mice fed the two soybean oil diets (plus those fed a high coconut oil and Viv chow diet). Our results show that the new high oleic soybean oil induces less obesity and adiposity than conventional soybean oil, but can cause hepatomegaly and liver dysfunction. Metabolomic analysis reveals that the hepatic and plasma metabolic profiles differ considerably between the two soybean oils. Hepatic C18 oxylipin metabolites of omega-6 (ω6) and omega-3 (ω3) fatty acids (linoleic and α-linolenic acid, respectively) in the cytochrome P450/soluble epoxide hydrolase pathway were found to correlate positively with obesity.
Institute:University of California, Riverside
Department:Cell Biology and Neuroscience
Last Name:Sladek
First Name:Frances
Address:2115 Biological Sciences Building,University of California, Riverside, CA 92521-0314
Email:frances.sladek@ucr.edu
Phone:951-827-2264

Subject:

Subject ID:SU000679
Subject Type:Animal
Subject Species:Mus musculus
Taxonomy ID:10090
Genotype Strain:C57/BL6N
Gender:Male
Animal Housing:SPF facility
Animal Light Cycle:12:12 h light-dark cycle
Animal Feed:Ad libitum
Animal Water:Ad libitum
Species Group:Mammal

Factors:

Subject type: Animal; Subject species: Mus musculus (Factor headings shown in green)

mb_sample_id local_sample_id GROUP_DESCRIPTION TIME_POINT Genotype
SA037647CSH_PM D4-16 e3_pos mode_MS_07212014.dHFD 24 weeks WT C57-Bl6
SA037648CSH_PM D4-16 e2_pos mode_MS_07212014.dHFD 24 weeks WT C57-Bl6
SA037649CSH_PM D4-16 e4_pos mode_MS_07212014.dHFD 24 weeks WT C57-Bl6
SA037650CSH_PM D4-17 e2_pos mode_MS_07212014.dHFD 24 weeks WT C57-Bl6
SA037651CSH_PM D4-18 e4_pos mode_MS_07212014.dHFD 24 weeks WT C57-Bl6
SA037652CSH_PM D4-16 e1_pos mode_MS_07212014.dHFD 24 weeks WT C57-Bl6
SA037653CSH_PM D4-17 e1_pos mode_MS_07212014.dHFD 24 weeks WT C57-Bl6
SA037654CSH_PM D4-18 e1_pos mode_MS_07212014.dHFD 24 weeks WT C57-Bl6
SA037655CSH_PM D3-17 e3reinject_pos mode_MS_07212014.dLA-HFD 24 weeks WT C57-Bl6
SA037656CSH_PM D3-17 e1_pos mode_MS_07212014.dLA-HFD 24 weeks WT C57-Bl6
SA037657CSH_PM D3-18 e3_pos mode_MS_07212014.dLA-HFD 24 weeks WT C57-Bl6
SA037658CSH_PM D3-17 e4_pos mode_MS_07212014.dLA-HFD 24 weeks WT C57-Bl6
SA037659CSH_PM D3-17 e2_pos mode_MS_07212014.dLA-HFD 24 weeks WT C57-Bl6
SA037660CSH_PM D3-18 e2reinject_pos mode_MS_07212014.dLA-HFD 24 weeks WT C57-Bl6
SA037661CSH_PM D3-18 e4_pos mode_MS_07212014.dLA-HFD 24 weeks WT C57-Bl6
SA037662CSH_PM D3-18 e1_pos mode_MS_07212014.dLA-HFD 24 weeks WT C57-Bl6
SA037663CSH_BioRec_02_pos mode_MS_07212014.dN/A N/A N/A
SA037664CSH_BioRec_03_pos mode_MS_07212014.dN/A N/A N/A
SA037665CSH_BioRec_01_pos mode_MS_07212014.dN/A N/A N/A
SA037666CSH_PM PL-5 e3_pos mode_MS_07212014.dPL-HFD 24 weeks WT C57-Bl6
SA037667CSH_PM PL-6 e4_pos mode_MS_07212014.dPL-HFD 24 weeks WT C57-Bl6
SA037668CSH_PM PL-6 e1_pos mode_MS_07212014.dPL-HFD 24 weeks WT C57-Bl6
SA037669CSH_PM PL-5 e1_pos mode_MS_07212014.dPL-HFD 24 weeks WT C57-Bl6
SA037670CSH_PM PL-5 e2_pos mode_MS_07212014.dPL-HFD 24 weeks WT C57-Bl6
SA037671CSH_PM PL-4 e3_pos mode_MS_07212014.dPL-HFD 24 weeks WT C57-Bl6
SA037672CSH_PM PL-4 e1_pos mode_MS_07212014.dPL-HFD 24 weeks WT C57-Bl6
SA037673CSH_PM PL-4 e4_pos mode_MS_07212014.dPL-HFD 24 weeks WT C57-Bl6
SA037674CSH_Viv-8_pos mode_MS_07212014.dViv chow 24 weeks WT C57-Bl6
SA037675CSH_Viv-9_pos mode_MS_07212014.dViv chow 24 weeks WT C57-Bl6
SA037676CSH_Viv-7_pos mode_MS_07212014.dViv chow 24 weeks WT C57-Bl6
SA037677CSH_Viv-4_pos mode_MS_07212014.dViv chow 24 weeks WT C57-Bl6
SA037678CSH_Viv-1_pos mode_MS_07212014.dViv chow 24 weeks WT C57-Bl6
SA037679CSH_Viv-2_pos mode_MS_07212014.dViv chow 24 weeks WT C57-Bl6
SA037680CSH_Viv-5_pos mode_MS_07212014.dViv chow 24 weeks WT C57-Bl6
SA037681CSH_Viv-6_pos mode_MS_07212014.dViv chow 24 weeks WT C57-Bl6
Showing results 1 to 35 of 35

Collection:

Collection ID:CO000673
Collection Summary:Liver tissue from mice on the diets for 24 weeks for metabolomic analysis was collected, rinsed in cold PBS, excess fluid was blotted with a kim-wipe and tissue was immediately snap frozen in liquid nitrogen before storage at -80°C. Blood was collected by cardiac puncture and centrifuged at 9 rcf for 5 min at 4°C. Plasma was stored immediately at -20°C.
Sample Type:Blood

Treatment:

Treatment ID:TR000693
Treatment Summary:Male C57/BL6N mice weaned at 3 weeks of age were randomly assigned to one of the four diets: 1) VIV chow: normal rodent chow, low in fat and high in fiber 2) HFD: 40 kcal% coconut oil 3) LA-HFD: 40 kcal% total fat soybean oil diet (21 kcal% from coconut oil and 19 kcal% from soybean oil) 4) PL-HFD: 40 kcal% total fat Plenish oil diet (21 kcal% from coconut oil and 19 kcal% from Plenish oil)

Sample Preparation:

Sampleprep ID:SP000686
Sampleprep Summary:1) Keep Specimen on Dry Ice 2) Transfer Tissue Contents into a new 1.5mL labeled eppendorf tube; keep on dry ice at all times 3) Add three (3) 3mm metal grinding balls to each sample; store in -80C for 10minutes 4) Homogenize the entire tissue to fine powder using genogrinder; make sure that the metal grinding balls are ice-cold prior to homogenization (step 3) 5) Upon completion of homogenization, keep samples on dry ice 6) Weight out two (2) aliquots: a ~5mg aliquot for CSH_lipidomics and a ~4mg aliquot for Primary Metabolites by GCTOF a. Record the exact weight weighed out for each sample b. Keep all samples on dry ice 7) KEEP remaining tissue specimen (>90mg) for analysis of Oxylipins (store in -80C) 8) Analysis of Primary Metabolites (GCTOFMS) a. Add 1mL of ice-cold “degassed” 3:3:2 ACN/IPA/H2O b. Vortex for 10seconds c. Shake on shaker for 20min at -4C d. Centrifuge the samples for 2min at 14,000 rcf e. Transfer two (2) 500μL aliquots to new 1.5mL eppendorf tubes; one for backup the other to be dried to dryness using the SpeedVac f. IMPORTANT: The precipitated protein will be used for analysis of the proteome, DO NOT DISCARD THESE; Place these in a separate labeled box and store in -20C g. Keep all samples on ice during extraction period h. Dry down one (1) 500μL aliquot to complete dryness i. Perform cleanup on dried aliquot using 500μL of 50/50 v/v ACN/H2O j. Transfer supernatant and dry to completeness k. Submit for Derivatization 9) Analysis of Complex Lipids (LCQTOF) a. Add 225μL of ice-cold “degassed” MeOH containing “ISTD mixture” to homogenized 5mg aliquot b. Vortex for 10 seconds c. Add 750μL of ice-cold “degassed” MTBE containing 22:1 CE ISTD d. Vortex for 10 seconds e. Shake on Orbital Mixer for 6min at 4C f. Add 188μL of room temperature H2O g. Vortex for 20 seconds h. Centrifuge for 2min at 14,000 rcf i. Transfer two (2) aliquots of 350μL of top layer, one for backup stored in -20C, the other for analysis j. Keep bottom layer and store in -20C k. Dry down one (1) 350μL aliquot to dryness using the Speedvac l. Resuspend samples in 108.6μL of 50ng/mL CUDA m. Vortex and sonicate for 5minutes n. Centrifuge for 2min at 14,000 rcf o. Transfer 90μL to an amber vial with micro-insert (non-diluted) p. Transfer 10μL to a new 1.5mL eppendorf tube, dilute 20X with 50ng/mL CUDA in 90:10 MeOH:Toluene (10μL + 190μL CUDA) and transfer 100μL to amber vial with micro-insert (diluted for TGs) i. The dilution is based off previous experiences with liver samples
Sampleprep Protocol Filename:SOP_Sample_preparation_of_blood_plasma_or_serum_samples_for_GCTOF_analysis.pdf

Combined analysis:

Analysis ID AN001001 AN001002
Analysis type MS MS
Chromatography type Reversed phase Reversed phase
Chromatography system Agilent 6530 Agilent 6550
Column Waters Acquity CSH C18 (100 x 2.1mm, 1.7um) Waters Acquity CSH C18 (100 x 2.1mm, 1.7um)
MS Type ESI ESI
MS instrument type QTOF QTOF
MS instrument name Agilent 6530 QTOF Agilent 6550 QTOF
Ion Mode POSITIVE NEGATIVE
Units Counts Counts

Chromatography:

Chromatography ID:CH000718
Methods Filename:Data_Dictionary_Fiehn_laboratory_CSH_QTOF_lipidomics_09-27-2013_general.pdf
Instrument Name:Agilent 6530
Column Name:Waters Acquity CSH C18 (100 x 2.1mm, 1.7um)
Column Pressure:450-850 bar
Column Temperature:65 C
Flow Gradient:15% B to 99% B
Flow Rate:0.6 mL/min
Injection Temperature:4 C
Internal Standard:See data dictionary
Retention Time:See data dictionary
Sample Injection:1.67 uL
Solvent A:60:40 Acetonitrile:Water +10mM Ammonium Formate +10mM Formic Acid
Solvent B:9:1 Isopropanol:Acetonitrile +10mM Ammonium Formate +10mM Formic Acid
Analytical Time:13 min
Capillary Voltage:3500 eV
Time Program:15 min
Weak Wash Solvent Name:Isopropanol
Strong Wash Solvent Name:Isopropanol
Target Sample Temperature:Autosampler temp 4 C
Randomization Order:Excel generated
Chromatography Type:Reversed phase
  
Chromatography ID:CH000719
Methods Filename:Data_Dictionary_Fiehn_laboratory_CSH_QTOF_lipidomics_09-27-2013_general.pdf
Instrument Name:Agilent 6550
Column Name:Waters Acquity CSH C18 (100 x 2.1mm, 1.7um)
Column Pressure:450-850 bar
Column Temperature:65 C
Flow Gradient:15% B to 99% B
Flow Rate:0.6 mL/min
Injection Temperature:4 C
Internal Standard:See data dictionary
Retention Time:See data dictionary
Sample Injection:5.0 uL
Solvent A:60:40 Acetonitrile:Water +10mM Ammonium Acetate +10mM Acetic Acid
Solvent B:9:1 Isopropanol:Acetonitrile +10mM Ammonium Acetate +10mM Acetic Acid
Analytical Time:13 min
Capillary Voltage:3500 eV
Time Program:15 min
Weak Wash Solvent Name:Isopropanol
Strong Wash Solvent Name:Isopropanol
Target Sample Temperature:Autosampler temp 4 C
Randomization Order:Excel
Chromatography Type:Reversed phase

MS:

MS ID:MS000896
Analysis ID:AN001001
Instrument Name:Agilent 6530 QTOF
Instrument Type:QTOF
MS Type:ESI
Ion Mode:POSITIVE
Capillary Voltage:3500 eV
Collision Energy:25 eV
Collision Gas:Nitrogen
Dry Gas Flow:8L/min
Dry Gas Temp:325 C
Fragment Voltage:120 eV
Fragmentation Method:Auto MS/MS
Ion Source Temperature:325 C
Ion Spray Voltage:1000
Ionization:Pos
Precursor Type:Intact Molecule
Reagent Gas:Nitrogen
Source Temperature:325 C
Dataformat:.d
Desolvation Gas Flow:11 L/min
Desolvation Temperature:350 C
Nebulizer:35 psig
Octpole Voltage:750
Resolution Setting:Exteded Dyamic Range
Scan Range Moverz:60-1700 Da
Scanning Cycle:2 Hz
Scanning Range:60-1700 Da
Skimmer Voltage:65
  
MS ID:MS000897
Analysis ID:AN001002
Instrument Name:Agilent 6550 QTOF
Instrument Type:QTOF
MS Type:ESI
Ion Mode:NEGATIVE
Capillary Voltage:3500 eV
Collision Energy:25 eV
Collision Gas:Nitrogen
Dry Gas Flow:13L/min
Dry Gas Temp:200 C
Fragment Voltage:175 eV
Fragmentation Method:Auto MS/MS
Ion Source Temperature:325 C
Ion Spray Voltage:1000
Ionization:Neg
Precursor Type:Intact Molecule
Reagent Gas:Nitrogen
Source Temperature:325 C
Dataformat:.d
Desolvation Gas Flow:11 L/min
Desolvation Temperature:350 C
Nebulizer:35 psig
Octpole Voltage:750
Resolution Setting:Exteded Dyamic Range
Scan Range Moverz:60-1700 Da
Scanning Cycle:2 Hz
Scanning Range:60-1700 Da
Skimmer Voltage:65
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