Summary of study ST000897

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 PR000623. The data can be accessed directly via it's Project DOI: 10.21228/M84T25 This work is supported by NIH grant, U2C- DK119886.

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Study IDST000897
Study TitleUntargeted metabolomics analysis of ischemia-reperfusion injured hearts ex vivo from sedentary and exercise trained rats.
Study SummaryThe effects of exercise on the heart and its resistance to disease are well-documented. Recent studies have identified exercise-induced resistance to arrhythmia is due to the preservation of mitochondrial membrane potential. To identify novel metabolic changes that occurred parallel to these mitochondrial alterations, we performed non-targeted metabolomics analysis on hearts from sedentary (Sed) and exercise- trained (Ex) rats challenged with isolated heart ischemia-reperfusion injury (I/R). Eight weeks old Sprague- Dawley rats were treadmill trained five days/week for six weeks (exercise duration and intensity progressively increased to 1 hour at 30 m/min up to 10.5% incline, 75-80% VO2mx). The recovery of pre-ischemic function for sedentary rat hearts was 28.8+/-5.4% (N=12) compared to exercise trained hearts which recovered 51.9%+/-5.7 (N=14)(p<0.001). Non-targeted GC-MS metabolomics analysis of 1) Sedentary rat hearts; 2) Exercise-trained rat hearts; 3) Sedentary rat hearts challenged with global ischemia-reperfusion (I/R) injury; and 4) Exercise-trained rat hearts challeged with global I/R (10/group) revealed 20 statistically significant metabolites between groups by ANOVA using Metaboanalyst (p<0.001). Enrichment analysis of these metabolites for pathway-associated metabolic sets indicated a >10 fold enrichment for ammonia recycling and protein biosynthesis (L-Glutamic acid; L-Proline; L-Histidine; L-Serine; L-Aspartic acid; L-Glutamine)(p<=4.05E-05, FDR=0.0024). Subsequent comparison of the sedentary hearts post-I/R and exercise-trained hearts post-I/R further identified significant differences in metabolites related to Aminoacyl-tRNA biosynthesis and nitrogen metabolism (4) (p<=1.24E-05, FDR<=5.07E-4). These studies shed light on novel mechanisms in which exercise-induced cardioprotection occurs in I/R which complement both the mitochondrial stabilization and antioxidant mechanisms recently described. These findings also link protein synthesis and protein degradation (protein quality control mechanisms) with exercise-linked cardioprotection and mitochondrial susceptibility for the first time in cardiac I/R.
Institute
University of North Carolina at Chapel Hill
DepartmentMcAllister heart Institute, Department of Internal medicine
LaboratoryMultiple Centers
Last NameWillis
First NameMonte
Address111 Mason Farm road, Chapel Hill, North Carolina, 27599-7126, USA
Emailmonte_willis@med.unc.edu
Phone919-360-7599
Submit Date2017-05-20
Study CommentsCardiac tissue
Raw Data AvailableYes
Raw Data File Type(s).d
Analysis Type DetailGC-MS
Release Date2018-02-07
Release Version1
Monte Willis Monte Willis
https://dx.doi.org/10.21228/M84T25
ftp://www.metabolomicsworkbench.org/Studies/ application/zip

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Project:

Project ID:PR000623
Project DOI:doi: 10.21228/M84T25
Project Title:Non targeted metabolomic profiling of sedentary vs. exercise trained rat hearts +/- ischemia reperfusion injury rats
Project Type:GC-MS non targeted analysis
Project Summary:Non targeted meatbolomic profiling of sedentary vs. exercise trained rat hearts +/- ischemia reperfusion injury
Institute:University of North Carolina at Chapel Hill
Department:McAllister heart Institute, Department of Pathology & Laboratory Medicine
Laboratory:Multiple Centers
Last Name:Willis
First Name:Monte
Address:111 Mason Farm road, Chapel Hill, North Carolina, 27599-7126, USA
Email:monte_willis@med.unc.edu
Phone:919-360-7599
Funding Source:NIH, Fondation Leducq

Subject:

Subject ID:SU000934
Subject Type:Animal
Subject Species:Rattus norvegicus
Taxonomy ID:10116
Species Group:Mammal

Factors:

Subject type: Animal; Subject species: Rattus norvegicus (Factor headings shown in green)

mb_sample_id local_sample_id Animal Group
SA052718H14Exercise Control
SA052719H13Exercise Control
SA052720H5Exercise Control
SA052721H18Exercise Control
SA052722H28Exercise Control
SA052723H39Exercise Control
SA052724H36Exercise Control
SA052725H2Exercise Control
SA052726H27Exercise Control
SA052727H23Exercise Control
SA052728H22Exercise Ischemia/Reperfusion Injury
SA052729H29Exercise Ischemia/Reperfusion Injury
SA052730H31Exercise Ischemia/Reperfusion Injury
SA052731H38Exercise Ischemia/Reperfusion Injury
SA052732H20Exercise Ischemia/Reperfusion Injury
SA052733H9Exercise Ischemia/Reperfusion Injury
SA052734H6Exercise Ischemia/Reperfusion Injury
SA052735H11Exercise Ischemia/Reperfusion Injury
SA052736H17Exercise Ischemia/Reperfusion Injury
SA052737H40Exercise Ischemia/Reperfusion Injury
SA052738H1Sedentary Control
SA052739H25Sedentary Control
SA052740H30Sedentary Control
SA052741H32Sedentary Control
SA052742H21Sedentary Control
SA052743H19Sedentary Control
SA052744H34Sedentary Control
SA052745H8Sedentary Control
SA052746H15Sedentary Control
SA052747H4Sedentary Control
SA052748H3Sedentary Ischemia/Reperfusion Injury
SA052749H12Sedentary Ischemia/Reperfusion Injury
SA052750H10Sedentary Ischemia/Reperfusion Injury
SA052751H7Sedentary Ischemia/Reperfusion Injury
SA052752H16Sedentary Ischemia/Reperfusion Injury
SA052753H24Sedentary Ischemia/Reperfusion Injury
SA052754H35Sedentary Ischemia/Reperfusion Injury
SA052755H33Sedentary Ischemia/Reperfusion Injury
SA052756H26Sedentary Ischemia/Reperfusion Injury
SA052757H37Sedentary Ischemia/Reperfusion Injury
Showing results 1 to 40 of 40

Collection:

Collection ID:CO000928
Collection Summary:Cardiac tissue was harvested amd then flash frozen in a liquid nitrogen.
Sample Type:Muscle

Treatment:

Treatment ID:TR000948
Treatment Summary:Fraction of cardiac 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 and placed on dry ice/stored at - 80C Fraction of cardiac 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 and placed on dry ice/stored at - 80C

Sample Preparation:

Sampleprep ID:SP000941
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. 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 AN001459
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 Peak values (Log transformed)

Chromatography:

Chromatography ID:CH001026
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:MS001347
Analysis ID:AN001459
Instrument Name:Agilent 5975
Instrument Type:Single quadrupole
MS Type:EI
Ion Mode:POSITIVE
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