Summary of Study ST000425
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 PR000332. The data can be accessed directly via it's Project DOI: 10.21228/M84G7M This work is supported by NIH grant, U2C- DK119886.
See: https://www.metabolomicsworkbench.org/about/howtocite.php
| Study ID | ST000425 |
| Study Title | Non targeted metabolomics of gastrocnemius tissue samples obtained from 20 month old (old) mice- Both Sham and after inducing lung injury (part I) |
| Study Type | Non targeted metabolomic analysis |
| Study Summary | Introduction: Older patients are more likely to acquire and die from acute respiratory distress syndrome (ARDS) and muscle weakness may be more significant in older survivors. Recent data implicate muscle ring finger protein 1 (MuRF1) in lung injury-induced skeletal muscle atrophy in young mice and identify an alternative role for MuRF1 in cardiac metabolism regulation through inhibition of fatty acid oxidation. Objectives: To develop a model of lung injury-induced muscle wasting in old mice and to evaluate the skeletal muscle metabolomic profile of adult and old acute lung injury (ALI) mice. Methods: Young (2 month), adult (6 month) and old (20 month) male C57Bl6J mice underwent Sham (intratracheal H2O) or ALI [intratracheal E. coli lipopolysaccharide (i.t. LPS)] conditions and muscle functional testing. Metabolomic analysis on gastrocnemius muscle was performed using gas chromatography-mass spectrometry (GC-MS). Results: Old ALI mice had increased mortality and failed to recover skeletal muscle function compared to adult ALI mice. Muscle MuRF1 expression was increased in old ALI mice at day 3. Non-targeted muscle metabolomics revealed alterations in amino acid biosynthesis and fatty acid metabolism in old ALI mice. Targeted metabolomics of fatty acid intermediates (acyl-carnitines) and amino acids revealed a reduction in long chain acyl-carnitines in old ALI mice. Conclusion: This study demonstrates age-associated susceptibility to ALI-induced muscle wasting which parallels a metabolomic profile suggestive of altered muscle fatty acid metabolism. MuRF1 activation may contribute to both atrophy and impaired fatty acid oxidation, which may synergistically impair muscle function in old ALI mice. |
| Institute | University of North Carolina;Duke University |
| Department | UNC McAllister Heart Institute;Duke Molecular Physiology Institute |
| Laboratory | Multiple Centers |
| Last Name | Ilaiwy;WIllis |
| First Name | Amro;Monte |
| Address | 111 Mason Farm road, Chapel Hill, North Carolina, 27599-7126, USA |
| monte_willis@med.unc.edu, amroilaiwy@gmail.com | |
| Phone | 210-596-0171 |
| Submit Date | 2016-06-21 |
| Analysis Type Detail | GC-MS |
| Release Date | 2016-09-23 |
| Release Version | 1 |
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Project:
| Project ID: | PR000332 |
| Project DOI: | doi: 10.21228/M84G7M |
| Project Title: | Lung injury-induced skeletal muscle wasting in aged mice is linked to alterations in long chain fatty acid metabolism |
| Project Type: | Metabolomics |
| Project Summary: | Non targeted and targeted metabolomic analysis on gastrocnemius tissue samples obtained from skeletal muscle of adult and old mice after inducing lung injury |
| Institute: | University of North Carolina at Chapel Hill |
| Department: | McAllister Heart Institute, Department of Internal Medicine |
| Laboratory: | Multiple Centers |
| Last Name: | Ilaiwy; Willis |
| First Name: | Amro; Monte |
| Address: | 111 Mason Farm road, Chapel Hill, North Carolina, 27599-7126, USA |
| Email: | monte_willis@med.unc.edu, amroilaiwy@gmail.com |
| Phone: | 210-596-0171 |
| Funding Source: | NIH, Fondation Leducq, Claude D. Pepper Older Americans Independence Center, the American Thoracic Society Foundation |
Subject:
| Subject ID: | SU000446 |
| Subject Type: | Animal |
| Subject Species: | Mus musculus |
| Taxonomy ID: | 10090 |
| Species Group: | Mammal |
Factors:
Subject type: Animal; Subject species: Mus musculus (Factor headings shown in green)
| mb_sample_id | local_sample_id | Group |
|---|---|---|
| SA021574 | A110 | ALI 20mth |
| SA021575 | A109 | ALI 20mth |
| SA021576 | A108 | ALI 20mth |
| SA021577 | A111 | ALI 20mth |
| SA021578 | A46 | ALI 20mth |
| SA021579 | A48 | ALI 20mth |
| SA021580 | A47 | ALI 20mth |
| SA021581 | A106 | ALI 20mth |
| SA021582 | A45 | ALI 20mth |
| SA021583 | A05 | ALI 20mth |
| SA021584 | A12 | Sham 20mth |
| SA021585 | A11 | Sham 20mth |
| SA021586 | A10 | Sham 20mth |
| SA021587 | A09 | Sham 20mth |
| SA021588 | A32 | Sham 20mth |
| SA021589 | A33 | Sham 20mth |
| SA021590 | A52 | Sham 20mth |
| SA021591 | A50 | Sham 20mth |
| SA021592 | A34 | Sham 20mth |
| SA021593 | A08 | Sham 20mth |
| Showing results 1 to 20 of 20 |
Collection:
| Collection ID: | CO000440 |
| Collection Summary: | Gastrocnemius tissue was harvested and then flash frozen in a liquid nitrogen cooled biopress |
| Sample Type: | Muscle |
Treatment:
| Treatment ID: | TR000460 |
| Treatment Summary: | Treatment Summary Fraction of gastrocnemius tissue weighed (25–50 mg wet weight), then the finely cut up tissue quickly added to fresh pre-made buffer (50 % acetyl-nitrile, 50 % water, 0.3% formic acid) at a standard concentration of 25 mg/475 mcl buffer then fully homogenized on ice for 10–25 s and placed on dry ice/stored at - 80C |
Sample Preparation:
| Sampleprep ID: | SP000453 |
| Sampleprep Summary: | The samples were crash deprotonized by methanol precipitation and spiked with D27-deuterated myristic acid (D27-C14:0) as an internal standard for retention-time locking and dried. The trimethylsilyl-D27-C14:0 standard retention time (RT) was set at 16.727 min. Reactive carbonyls were stabilized at 50C with methoxyamine hydrochloride in dry pyridine. Metabolites were made volatile with TMS groups using N-methyl-N-(trimethylsilyl) trifluoroacetamide or MSTFA with catalytic trimethylchlorosilane at 50C. |
Combined analysis:
| Analysis ID | AN000675 |
|---|---|
| Analysis type | MS |
| Chromatography type | GC |
| Chromatography system | Agilent 6890N |
| Column | Agilent DB5-MS (15m x 0.25mm,0.25um) |
| MS Type | EI |
| MS instrument type | Single quadrupole |
| MS instrument name | Agilent 5975 |
| Ion Mode | POSITIVE |
| Units | Log transformed peak values |
Chromatography:
| Chromatography ID: | CH000487 |
| Chromatography Summary: | GC/MS methods follow previous studies using a 6890 N GC connected to a 5975 Inert single quadrupole MS (Agilent Technologies, Santa Clara, CA) (Bonikos et al. 1975; Fiehn 2008; Kind et al. 2009). The two wall-coated, open-tubular GC columns connected in series are both from J&W/Agilent (part 122–5512), DB5-MS, 15 meters in length, 0.25 mm in diameter, with an 0.25-l m luminal film. Positive ions generated with conventional electron-ionization at 70 eV are scanned broadly from 600 to 50 m/z in the detector throughout the 45 min cycle time. |
| Instrument Name: | Agilent 6890N |
| Column Name: | Agilent DB5-MS (15m x 0.25mm,0.25um) |
| Chromatography Type: | GC |
MS:
| MS ID: | MS000601 |
| Analysis ID: | AN000675 |
| Instrument Name: | Agilent 5975 |
| Instrument Type: | Single quadrupole |
| MS Type: | EI |
| Ion Mode: | POSITIVE |
