Summary of study ST000426

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 IDST000426
Study TitleNon targeted metabolomics of gastrocnemius tissue samples obtained from 6 month old (adult) mice- Both Sham and after inducing lung injury (part II)
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 Date2016-06-30
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:SU000447
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
SA021594M39ALI 6mth
SA021595M38ALI 6mth
SA021596M37ALI 6mth
SA021597M62ALI 6mth
SA021598M64ALI 6mth
SA021599M66ALI 6mth
SA021600M65ALI 6mth
SA021601M36ALI 6mth
SA021602M63ALI 6mth
SA021603M35ALI 6mth
SA021604M29Sham 6mth
SA021605M28Sham 6mth
SA021606M12Sham 6mth
SA021607M11Sham 6mth
SA021608M30Sham 6mth
SA021609M53Sham 6mth
SA021610M9Sham 6mth
SA021611M55Sham 6mth
SA021612M54Sham 6mth
SA021613M10Sham 6mth
Showing results 1 to 20 of 20

Collection:

Collection ID:CO000441
Collection Summary:Gastrocnemius tissue was harvested and then flash frozen in a liquid nitrogen cooled biopress
Sample Type:Muscle

Treatment:

Treatment ID:TR000461
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:SP000454
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 AN000676
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:CH000488
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:MS000602
Analysis ID:AN000676
Instrument Name:Agilent 5975
Instrument Type:Single quadrupole
MS Type:EI
Ion Mode:POSITIVE
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