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.

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Study IDST000425
Study TitleNon targeted metabolomics of gastrocnemius tissue samples obtained from 20 month old (old) mice- Both Sham and after inducing lung injury (part I)
Study TypeNon targeted metabolomic analysis
Study SummaryIntroduction: 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
DepartmentUNC McAllister Heart Institute;Duke Molecular Physiology Institute
LaboratoryMultiple Centers
Last NameIlaiwy;WIllis
First NameAmro;Monte
Address111 Mason Farm road, Chapel Hill, North Carolina, 27599-7126, USA
Emailmonte_willis@med.unc.edu, amroilaiwy@gmail.com
Phone210-596-0171
Submit Date2016-06-21
Analysis Type DetailGC-MS
Release Date2016-09-23
Release Version1
Amro Ilaiwy Amro Ilaiwy
Monte WIllis Monte WIllis
https://dx.doi.org/10.21228/M84G7M
ftp://www.metabolomicsworkbench.org/Studies/ application/zip

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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
SA021574A110ALI 20mth
SA021575A109ALI 20mth
SA021576A108ALI 20mth
SA021577A111ALI 20mth
SA021578A46ALI 20mth
SA021579A48ALI 20mth
SA021580A47ALI 20mth
SA021581A106ALI 20mth
SA021582A45ALI 20mth
SA021583A05ALI 20mth
SA021584A12Sham 20mth
SA021585A11Sham 20mth
SA021586A10Sham 20mth
SA021587A09Sham 20mth
SA021588A32Sham 20mth
SA021589A33Sham 20mth
SA021590A52Sham 20mth
SA021591A50Sham 20mth
SA021592A34Sham 20mth
SA021593A08Sham 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 × 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 × 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
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