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.
See: https://www.metabolomicsworkbench.org/about/howtocite.php
| Study ID | ST000401 |
| Study Title | Inhibition of diamine oxidase promotes uptake of putrescine from rat small intestine |
| Study 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 |
| ofiehn@ucdavis.edu | |
| Phone | (530) 754-8258 |
| Submit Date | 2016-05-16 |
| Num Groups | 5 |
| Total Subjects | 40 |
| Raw Data Available | Yes |
| Raw Data File Type(s) | peg |
| Analysis Type Detail | GC-MS |
| Release Date | 2016-06-18 |
| Release Version | 1 |
Select appropriate tab below to view additional metadata details:
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 |
|---|---|---|---|
| SA019215 | 140603actsa30_1 | Duodenum | Aminoguanidine 10 mg/kg s.c. |
| SA019216 | 140603actsa25_1 | Duodenum | Aminoguanidine 10 mg/kg s.c. |
| SA019217 | 140603actsa27_1 | Duodenum | Aminoguanidine 10 mg/kg s.c. |
| SA019218 | 140603actsa06_1 | Duodenum | Aminoguanidine 10 mg/kg s.c. |
| SA019219 | 140603actsa17_1 | Duodenum | Metformin 100mg/kg (oral administration) |
| SA019220 | 140603actsa35_1 | Duodenum | Metformin 100mg/kg (oral administration) |
| SA019221 | 140603actsa07_1 | Duodenum | Metformin 100mg/kg (oral administration) |
| SA019222 | 140603actsa03_1 | Duodenum | Metformin 100mg/kg (oral administration) |
| SA019223 | 140603actsa20_1 | Duodenum | Metformin 25mg/kg (oral administration) |
| SA019224 | 140603actsa18_1 | Duodenum | Metformin 25mg/kg (oral administration) |
| SA019225 | 140603actsa29_1 | Duodenum | Metformin 25mg/kg (oral administration) |
| SA019226 | 140603actsa33_1 | Duodenum | Metformin 25mg/kg (oral administration) |
| SA019227 | 140603actsa13_1 | Duodenum | Metformin 50mg/kg (oral administration) |
| SA019228 | 140603actsa32_3 | Duodenum | Metformin 50mg/kg (oral administration) |
| SA019229 | 140603actsa16_1 | Duodenum | Metformin 50mg/kg (oral administration) |
| SA019230 | 140603actsa38_1 | Duodenum | Metformin 50mg/kg (oral administration) |
| SA019231 | 140603actsa40_1 | Duodenum | Saline (oral administration) |
| SA019232 | 140603actsa31_1 | Duodenum | Saline (oral administration) |
| SA019233 | 140603actsa10_1 | Duodenum | Saline (oral administration) |
| SA019234 | 140603actsa24_1 | Duodenum | Saline (oral administration) |
| SA019235 | 140603actsa11_1 | Ileum | Aminoguanidine 10 mg/kg s.c. |
| SA019236 | 140603actsa05_1 | Ileum | Aminoguanidine 10 mg/kg s.c. |
| SA019237 | 140603actsa22_1 | Ileum | Aminoguanidine 10 mg/kg s.c. |
| SA019238 | 140603actsa02_1 | Ileum | Aminoguanidine 10 mg/kg s.c. |
| SA019239 | 140603actsa26_1 | Ileum | Metformin 100mg/kg (oral administration) |
| SA019240 | 140603actsa19_1 | Ileum | Metformin 100mg/kg (oral administration) |
| SA019241 | 140603actsa37_1 | Ileum | Metformin 100mg/kg (oral administration) |
| SA019242 | 140603actsa36_1 | Ileum | Metformin 100mg/kg (oral administration) |
| SA019243 | 140603actsa01_1 | Ileum | Metformin 25mg/kg (oral administration) |
| SA019244 | 140603actsa08_1 | Ileum | Metformin 25mg/kg (oral administration) |
| SA019245 | 140603actsa14_1 | Ileum | Metformin 25mg/kg (oral administration) |
| SA019246 | 140603actsa09_1 | Ileum | Metformin 25mg/kg (oral administration) |
| SA019247 | 140603actsa23_1 | Ileum | Metformin 50mg/kg (oral administration) |
| SA019248 | 140603actsa12_1 | Ileum | Metformin 50mg/kg (oral administration) |
| SA019249 | 140603actsa34_1 | Ileum | Metformin 50mg/kg (oral administration) |
| SA019250 | 140603actsa39_1 | Ileum | Metformin 50mg/kg (oral administration) |
| SA019251 | 140603actsa28_1 | Ileum | Saline (oral administration) |
| SA019252 | 140603actsa21_1 | Ileum | Saline (oral administration) |
| SA019253 | 140603actsa04_1 | Ileum | Saline (oral administration) |
| SA019254 | 140603actsa15_1 | Ileum | 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 500l 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 |
