Summary of study ST000401

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 PR000313. The data can be accessed directly via it's Project DOI: 10.21228/M8QG7W This work is supported by NIH grant, U2C- DK119886.

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Study IDST000401
Study TitleInhibition of diamine oxidase promotes uptake of putrescine from rat small intestine
Study SummaryMetformin, a biguanide molecule, which is used as first line therapy for type 2 diabetes. In this study, we would like to investigate the inhibition of an enzyme called diamine oxidase (DAO) (also known as ABP1), by metformin. Based on our preliminary in vitro study using diamine oxidase enzyme, we saw increased level of putrescine with increasing metformin concentrations (see reference PMID: 26335661). This proposed in vivo study was to determine whether metformin could increase putrescine levels and other metabolites in mice. Aminoguanidine, a known inhibitor of DAO, in this study as positive control, following similar study design described in this paper (PMID: 8912017).
Institute
University of California, Davis
DepartmentGenome and Biomedical Sciences Facility
LaboratoryWCMC Metabolomics Core
Last NameFiehn
First NameOliver
Address1315 Genome and Biomedical Sciences Facility, 451 Health Sciences Drive, Davis, CA 95616
Emailofiehn@ucdavis.edu
Phone(530) 754-8258
Submit Date2016-05-16
Num Groups5
Total Subjects40
Raw Data AvailableYes
Raw Data File Type(s).peg
Analysis Type DetailGC-MS
Release Date2016-06-18
Release Version1
Oliver Fiehn Oliver Fiehn
https://dx.doi.org/10.21228/M8QG7W
ftp://www.metabolomicsworkbench.org/Studies/ application/zip

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Project:

Project ID:PR000313
Project DOI:doi: 10.21228/M8QG7W
Project Title:Inhibition of diamine oxidase promotes uptake of putrescine from rat small intestine
Project Summary:Metformin, a biguanide molecule, which is used as first line therapy for type 2 diabetes. In this study, we would like to investigate the inhibition of an enzyme called diamine oxidase (DAO) (also known as ABP1), by metformin. Based on our preliminary in vitro study using diamine oxidase enzyme, we saw increased level of putrescine with increasing metformin concentrations (see reference PMID: 26335661). This proposed in vivo study was to determine whether metformin could increase putrescine levels and other metabolites in mice. Aminoguanidine, a known inhibitor of DAO, in this study as positive control, following similar study design described in this paper (PMID: 8912017).
Institute:University of California, Davis
Department:Genome and Biomedical Sciences Facility
Laboratory:WCMC Metabolomics Core
Last Name:Fiehn
First Name:Oliver
Address:1315 Genome and Biomedical Sciences Facility, 451 Health Sciences Drive, Davis, CA 95616
Email:ofiehn@ucdavis.edu
Phone:(530) 754-8258
Funding Source:NIH U24DK097154

Subject:

Subject ID:SU000422
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 Organ Treatment
SA019215140603actsa30_1Duodenum Aminoguanidine 10 mg/kg s.c.
SA019216140603actsa25_1Duodenum Aminoguanidine 10 mg/kg s.c.
SA019217140603actsa27_1Duodenum Aminoguanidine 10 mg/kg s.c.
SA019218140603actsa06_1Duodenum Aminoguanidine 10 mg/kg s.c.
SA019219140603actsa17_1Duodenum Metformin 100mg/kg (oral administration)
SA019220140603actsa35_1Duodenum Metformin 100mg/kg (oral administration)
SA019221140603actsa07_1Duodenum Metformin 100mg/kg (oral administration)
SA019222140603actsa03_1Duodenum Metformin 100mg/kg (oral administration)
SA019223140603actsa20_1Duodenum Metformin 25mg/kg (oral administration)
SA019224140603actsa18_1Duodenum Metformin 25mg/kg (oral administration)
SA019225140603actsa29_1Duodenum Metformin 25mg/kg (oral administration)
SA019226140603actsa33_1Duodenum Metformin 25mg/kg (oral administration)
SA019227140603actsa13_1Duodenum Metformin 50mg/kg (oral administration)
SA019228140603actsa32_3Duodenum Metformin 50mg/kg (oral administration)
SA019229140603actsa16_1Duodenum Metformin 50mg/kg (oral administration)
SA019230140603actsa38_1Duodenum Metformin 50mg/kg (oral administration)
SA019231140603actsa40_1Duodenum Saline (oral administration)
SA019232140603actsa31_1Duodenum Saline (oral administration)
SA019233140603actsa10_1Duodenum Saline (oral administration)
SA019234140603actsa24_1Duodenum Saline (oral administration)
SA019235140603actsa11_1Ileum Aminoguanidine 10 mg/kg s.c.
SA019236140603actsa05_1Ileum Aminoguanidine 10 mg/kg s.c.
SA019237140603actsa22_1Ileum Aminoguanidine 10 mg/kg s.c.
SA019238140603actsa02_1Ileum Aminoguanidine 10 mg/kg s.c.
SA019239140603actsa26_1Ileum Metformin 100mg/kg (oral administration)
SA019240140603actsa19_1Ileum Metformin 100mg/kg (oral administration)
SA019241140603actsa37_1Ileum Metformin 100mg/kg (oral administration)
SA019242140603actsa36_1Ileum Metformin 100mg/kg (oral administration)
SA019243140603actsa01_1Ileum Metformin 25mg/kg (oral administration)
SA019244140603actsa08_1Ileum Metformin 25mg/kg (oral administration)
SA019245140603actsa14_1Ileum Metformin 25mg/kg (oral administration)
SA019246140603actsa09_1Ileum Metformin 25mg/kg (oral administration)
SA019247140603actsa23_1Ileum Metformin 50mg/kg (oral administration)
SA019248140603actsa12_1Ileum Metformin 50mg/kg (oral administration)
SA019249140603actsa34_1Ileum Metformin 50mg/kg (oral administration)
SA019250140603actsa39_1Ileum Metformin 50mg/kg (oral administration)
SA019251140603actsa28_1Ileum Saline (oral administration)
SA019252140603actsa21_1Ileum Saline (oral administration)
SA019253140603actsa04_1Ileum Saline (oral administration)
SA019254140603actsa15_1Ileum Saline (oral administration)
Showing results 1 to 40 of 40

Collection:

Collection ID:CO000416
Collection Summary:After 1 hour of treatment, blood was collected from the portal vein. Blood from the portal vein was cannulated with a fine syringe and blood was aspirated. Blood was spun down and plasma was collected and stored in -80. Urine was collected from bladder after 1 hour. Tissues (liver, kidney, intestine) were collected at the end of 1 hour. Only duodenum and ileum were used for metabolomics analysis. This study was conducted under the auspices of approved IACUC procedures in MuriGenics, Inc (Vallejo, CA).
Sample Type:Tissue
Collection Location:MuriGenics, Inc (Vallejo, CA)
Collection Time:1 hour after treatment

Treatment:

Treatment ID:TR000436
Treatment Summary:Twenty (20) adult rats (male, weighing approximately 200g) were randomized by body weights and assigned to five treatment groups (saline, aminoguanidine (positive control), 3 different oral doses of metformin). Before treatment, rats were starved for 24 hours. On treatment day, saline or aminoguanidine or metformin will be given to the rat.
Animal Fasting:24 hours

Sample Preparation:

Sampleprep ID:SP000429
Sampleprep Summary:1. Weigh 4 mg tissue sample ( muscle 20 mg) in to a 2.0 ml eppendorf tube. Add 1.0 mL extraction solvent to the tissue sample and using GenoGrinder homogenize samples for 45 seconds ensuring that sample resembles a powder. 2. Centrifuge the samples at 2500 rpm. for 5 minutes. Aliquot 2 X 500µl supernatant, one for analysis and one for a backup sample. Store backup aliquot in the -20°C freezer. 3. Evaporate one 500µl aliquot of the sample in the Labconco Centrivap cold trap concentrator to complete dryness 4. The dried aliquot is then re-suspended with 500l 50% acetonitrile (degassed as given) (only for liver and brain samples). 5. Centrifuge for 2 min at 14000 rcf using the centrifuge Eppendorf 5415. 6. Remove supernatant to a new Eppendorff tube. 7. Evaporate the supernatant to dryness in the the Labconco Centrivap cold trap concentrator. 8. Submit to derivatization.
Sampleprep Protocol Filename:SOP_Extraction_of_Mammalian_Tissue_Samples.pdf

Combined analysis:

Analysis ID AN000640
Analysis type MS
Chromatography type GC
Chromatography system Agilent 6890N
Column Restek Corporation Rtx-5Sil MS
MS Type EI
MS instrument type GC-TOF
MS instrument name Leco Pegasus III GC TOF
Ion Mode POSITIVE
Units counts

Chromatography:

Chromatography ID:CH000465
Methods Filename:Data_Dictionary_Fiehn_laboratory_GCTOF_MS_primary_metabolism_10-15-2013_general.pdf
Instrument Name:Agilent 6890N
Column Name:Restek Corporation Rtx-5Sil MS
Column Pressure:7.7 PSI
Column Temperature:50-330C
Flow Rate:1 ml/min
Injection Temperature:50 C ramped to 250 C by 12 C/s
Sample Injection:0.5 uL
Oven Temperature:50°C for 1 min, then ramped at 20°C/min to 330°C, held constant for 5 min
Transferline Temperature:230C
Washing Buffer:Ethyl Acetate
Sample Loop Size:30 m length x 0.25 mm internal diameter
Randomization Order:Excel generated
Chromatography Type:GC

MS:

MS ID:MS000572
Analysis ID:AN000640
Instrument Name:Leco Pegasus III GC TOF
Instrument Type:GC-TOF
MS Type:EI
Ion Mode:POSITIVE
Ion Source Temperature:250 C
Ionization Energy:70 eV
Mass Accuracy:Nominal
Source Temperature:250 C
Scan Range Moverz:85-500 Da
Scanning Cycle:17 Hz
Scanning Range:85-500 Da
Skimmer Voltage:1850 V
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