Summary of Study ST001145
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 PR000765. The data can be accessed directly via it's Project DOI: 10.21228/M8SQ4S 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.
| Study ID | ST001145 |
| Study Title | UPLC-MS Analysis of Lipids From Insulin Resistant Femoral Muscles of Diet-induced Obese Mice |
| Study Type | Lipidomics, Basic Research |
| Study Summary | Muscle insulin resistance is a fundamental contributor in the pathogenesis of obesity-related diseases like type 2 diabetes. Increased triglyceride concentration in muscle tissue, as seen with obesity, is associated with inhibition of insulin action and decreased glucose uptake. Here we use liquid chromatography paired with mass spectrometry (LCMS) to identify patterns of lipid species in femoral muscle of mice associated with diet-induced insulin resistance. Mice were fed a standard CHOW diet for 5 weeks or HFD for 5 or 13 weeks. 806 lipids were significantly different (p ≤ 0.05) between HFD-induced insulin resistant muscle and CHOW insulin sensitive. Of these 217 lipid species were quantified and annotated based on principle components analysis, significance (p ≤ 0.01) and fold change of relative abundance values. CHOW insulin sensitive muscle was associated with triglycerides and phospholipids that contained higher abundance of long-chain highly unsaturated fatty acids. Serine and inositol phospholipids favored insulin sensitive femoral muscle, yet higher abundance also occurred in 13 week HFD mice compared with 5 week. Consequently, phospholipid imbalance may be indicative of cell membrane dysfunction. HFD insulin resistant femoral muscle contained triglycerides with less carbons, compared with CHOW, which were predominantly saturated. In addition, there was greater abundance of diacylglycerides and sphingomyelin, but not ceramides. Extending HFD intake to 13 weeks did not cause increased abundance of deleterious lipids with the exception of sphingomyelin. Overall, distinct lipid combinations, perhaps even ratios, should be characterized when identifying what contributes to the maintenance or dysregulation of muscle insulin sensitivity. |
| Institute | Colorado State University |
| Department | Food Science and Human Nutrition |
| Laboratory | Adipose Tissue |
| Last Name | Foster |
| First Name | Michelle |
| Address | 1571 Campus Delivery, Fort Collins, Colorado 80523 |
| Michelle.Foster@colostate.edu | |
| Phone | 9704916189 |
| Submit Date | 2019-01-18 |
| Num Groups | 3 |
| Total Subjects | 21 |
| Num Males | 21 |
| Raw Data Available | Yes |
| Raw Data File Type(s) | cdf |
| Analysis Type Detail | LC-MS |
| Release Date | 2020-01-06 |
| Release Version | 1 |
Select appropriate tab below to view additional metadata details:
Project:
| Project ID: | PR000765 |
| Project DOI: | doi: 10.21228/M8SQ4S |
| Project Title: | UPLC-MS Analysis of Lipids From Insulin Resistant Femoral Muscles of Diet-induced Obese Mice |
| Project Type: | Lipidomics |
| Project Summary: | Muscle insulin resistance is a fundamental contributor in the pathogenesis of obesity-related diseases like type 2 diabetes. Increased triglyceride concentration in muscle tissue, as seen with obesity, is associated with inhibition of insulin action and decreased glucose uptake. Here we use liquid chromatography paired with mass spectrometry (LCMS) to identify patterns of lipid species in femoral muscle of mice associated with diet-induced insulin resistance. Mice were fed a standard CHOW diet for 5 weeks or HFD for 5 or 13 weeks. 806 lipids were significantly different (p ≤ 0.05) between HFD-induced insulin resistant muscle and CHOW insulin sensitive. Of these 217 lipid species were quantified and annotated based on principle components analysis, significance (p ≤ 0.01) and fold change of relative abundance values. CHOW insulin sensitive muscle was associated with triglycerides and phospholipids that contained higher abundance of long-chain highly unsaturated fatty acids. Serine and inositol phospholipids favored insulin sensitive femoral muscle, yet higher abundance also occurred in 13 week HFD mice compared with 5 week. Consequently, phospholipid imbalance may be indicative of cell membrane dysfunction. HFD insulin resistant femoral muscle contained triglycerides with less carbons, compared with CHOW, which were predominantly saturated. In addition, there was greater abundance of diacylglycerides and sphingomyelin, but not ceramides. Extending HFD intake to 13 weeks did not cause increased abundance of deleterious lipids with the exception of sphingomyelin. Overall, distinct lipid combinations, perhaps even ratios, should be characterized when identifying what contributes to the maintenance or dysregulation of muscle insulin sensitivity. |
| Institute: | Colorado State University |
| Department: | Food Science and Human Nutrition |
| Laboratory: | Adipose Tissue |
| Last Name: | Foster |
| First Name: | Michelle |
| Address: | 1571 Campus Delivery, Fort Collins Colorado |
| Email: | Michelle.Foster@colostate.edu |
| Phone: | 970-491-6189 |
| Funding Source: | NIH NIDDK |
Subject:
| Subject ID: | SU001210 |
| Subject Type: | Mammal |
| Subject Species: | Mus musculus |
| Taxonomy ID: | 10090 |
| Genotype Strain: | C57BL6 |
| Age Or Age Range: | 2-3 months |
| Weight Or Weight Range: | 28-46 |
| Gender: | Female |
| Animal Animal Supplier: | Jackson Laboratory |
| Animal Housing: | Single House |
| Animal Light Cycle: | 12:12 |
| Animal Feed: | CHOW diet (Envigo Teklad 6% fat 7002, Madison, WI) and Western (HFD; high-fat, high-sugar; 21% milk fat and 34% sucrose (Envigo TD.08811) |
Factors:
Subject type: Mammal; Subject species: Mus musculus (Factor headings shown in green)
| mb_sample_id | local_sample_id | Diet | Time |
|---|---|---|---|
| SA079564 | 19 | CHOW | 13week |
| SA079565 | 9 | CHOW | 13week |
| SA079566 | 16 | CHOW | 13week |
| SA079567 | 12 | CHOW | 13week |
| SA079568 | 8 | CHOW | 13week |
| SA079569 | 21 | CHOW | 13week |
| SA079570 | 2 | CHOW | 13week |
| SA079571 | 3 | CHOW | 13week |
| SA079572 | 5 | CHOW | 13week |
| SA079573 | 15 | CHOW | 13week |
| SA079574 | 17 | HFD | 5week |
| SA079575 | 20 | HFD | 5week |
| SA079576 | 18 | HFD | 5week |
| SA079577 | 11 | HFD | 5week |
| SA079578 | 6 | HFD | 5week |
| SA079579 | 4 | HFD | 5week |
| SA079580 | 7 | HFD | 5week |
| SA079581 | 10 | HFD | 5week |
| SA079582 | 13 | HFD | 5week |
| SA079583 | 1 | HFD | 5week |
| SA079584 | 14 | HFD | 5week |
| Showing results 1 to 21 of 21 |
Collection:
| Collection ID: | CO001204 |
| Collection Summary: | Approximately 20 mg of muscle tissue was homogenized in a glass homogenizer with 1.5 ml of 2:1 chloroform:methanol and then brought to 4 ml using the same ratio. The mixture was poured through a 2V grade qualitative 12.5 cm Whatman filter into a clean 10 ml glass tube. The volume in the tube was again brought up to 4 ml with the same 2:1 solution as above. One ml of water was added to the tube, vortexed for 20 seconds, and then centrifuged for 10 minutes at 2500 rpm. The top non-lipid portion was removed and the lower lipid-containing layer was dried under nitrogen. |
| Sample Type: | Muscle |
| Collection Method: | excision |
| Collection Location: | femoral muscle |
| Storage Conditions: | -80℃ |
Treatment:
| Treatment ID: | TR001225 |
| Treatment Summary: | Male C57BL/6 mice, 3 months of age, (Jackson Laboratory, Bar Harbor, Maine) were allowed to acclimate for one week before experiment start. Mice were individually housed under controlled conditions (12:12 light-dark cycle, 50–60% humidity, and 25° C) and had ad libitum access to standard CHOW diet (Envigo Teklad 6% fat 7002, Madison, WI). Lipids in the CHOW diet consisted of an assortment of fatty acids where linoleic > oleic > palmitic > linolenic > stearic. Following a baseline glucose tolerance test (GTT), mice were grouped according to mean GTT and body mass into a standard 5 week CHOW (n = 10) or Western (HFD; high-fat, high-sugar; 21% milk fat and 34% sucrose (Envigo TD.08811); 5 (n = 5) and 13 week (n = 6)) diet group. The saturated fatty acids in HFD ranged from 4:0 to 18:0, however, palmitate (16:0) followed by steric (18:0) and myristic (14;0) where highest in quantity. |
| Treatment: | Diet |
| Treatment Compound: | Envigo TD.08811 |
| Treatment Route: | oral |
Sample Preparation:
| Sampleprep ID: | SP001218 |
| Sampleprep Summary: | Approximately 20 mg of muscle tissue was homogenized in a glass homogenizer with 1.5 ml of 2:1 chloroform:methanol and then brought to 4 ml using the same ratio. The mixture was poured through a 2V grade qualitative 12.5 cm Whatman filter into a clean 10 ml glass tube. The volume in the tube was again brought up to 4 ml with the same 2:1 solution as above. One ml of water was added to the tube, vortexed for 20 seconds, and then centrifuged for 10 minutes at 2500 rpm. The top non-lipid portion was removed and the lower lipid-containing layer was dried under nitrogen.Lipid extracts were suspended in 100 uL of 2:1 Chloroform:Methanol. Injections were normalized such that equal amounts of lipid were analyzed for each sample, regardless of total lipid content of diet. 3 μL of extract was injected twice (n=2 replicates) onto a Waters Acquity UPLC system in discrete, randomized blocks. Next samples were separated using a Waters Acquity UPLC CSH Phenyl Hexyl column (1.7 µM, 1.0 x 100 mm), using a gradient from solvent A (water, 0.1% formic acid) to solvent B (Acetonitrile, 0.1% formic acid). Injections were made in 100% A, held at 100% A for 1 min, ramped to 98% B over 12 minutes, held at 98% B for 3 minutes, and then returned to starting conditions over 0.05 minutes and allowed to re-equilibrate for 3.95 minutes, with a 200 µL/min constant flow rate. The column and samples were held at 65 °C and 6 °C, respectively. The column eluent was infused into a Waters Xevo G2 TOF-MS with an electrospray source in positive mode, scanning 50-2000 m/z at 0.2 seconds per scan, alternating between MS (6 V collision energy) and MSE mode (15-30 V ramp). Calibration was performed using sodium iodide with 1 ppm mass accuracy. The capillary voltage was held at 2200 V, source temp at 150 °C, and nitrogen desolvation temp at 350 °C with a flow rate of 800 L/hr. |
| Processing Storage Conditions: | -80℃ |
| Extract Storage: | On ice |
Combined analysis:
| Analysis ID | AN001890 |
|---|---|
| Analysis type | MS |
| Chromatography type | Reversed phase |
| Chromatography system | Waters Acquity UPLC |
| Column | Acquity CSH PhenylHexyl |
| MS Type | ESI |
| MS instrument type | QTOF |
| MS instrument name | Waters Synapt G2 XS QTOF |
| Ion Mode | POSITIVE |
| Units | Relative Abundance |
Chromatography:
| Chromatography ID: | CH001367 |
| Instrument Name: | Waters Acquity UPLC |
| Column Name: | Acquity CSH PhenylHexyl |
| Flow Gradient: | water + 0.1% Formic + 2 mM AmOH / Acetonitrile |
| Flow Rate: | 200 uL/min |
| Solvent A: | 100% water; 0.1% formic acid; 2 mM ammonium hydroxide |
| Solvent B: | 100% acetonitrile |
| Chromatography Type: | Reversed phase |
MS:
| MS ID: | MS001746 |
| Analysis ID: | AN001890 |
| Instrument Name: | Waters Synapt G2 XS QTOF |
| Instrument Type: | QTOF |
| MS Type: | ESI |
| MS Comments: | Binary Data Format: .cdf |
| Ion Mode: | POSITIVE |
