#METABOLOMICS WORKBENCH Ming2022_20220424_090446 DATATRACK_ID:3216 STUDY_ID:ST002156 ANALYSIS_ID:AN003531 PROJECT_ID:PR001369 VERSION 1 CREATED_ON May 3, 2022, 11:39 am #PROJECT PR:PROJECT_TITLE Contribution of western diet to NASH pathogenesis by defining liver metabolites PR:PROJECT_SUMMARY Western diet (WD) and gut microbiota interplay drives the development of PR:PROJECT_SUMMARY non-alcoholic fatty liver disease (NAFLD) and its progression to non-alcoholic PR:PROJECT_SUMMARY steatohepatitis (NASH). However, the metabolic mediators contributing to NASH PR:PROJECT_SUMMARY remain to be identified. In this study, we explored the change of liver PR:PROJECT_SUMMARY metabolites in a diet-induced mouse NASH model. Identification of these PR:PROJECT_SUMMARY metabolites will help to uncover the pathogenesis of NASH and explore the new PR:PROJECT_SUMMARY therapeutic treatment. PR:INSTITUTE University of Missouri-Columbia PR:LAST_NAME Yang PR:FIRST_NAME Ming PR:ADDRESS University of Missouri, NextGen Prevision Health Building, Room 2203 PR:EMAIL yangmin@health.missouri.edu PR:PHONE 573-882-7141 #STUDY ST:STUDY_TITLE Mouse liver metabolites ST:STUDY_SUMMARY Western diet (WD) and gut microbiota interplay drives the development of ST:STUDY_SUMMARY non-alcoholic fatty liver disease (NAFLD) and its progression to non-alcoholic ST:STUDY_SUMMARY steatohepatitis (NASH). However, the metabolic mediators contributing to NASH ST:STUDY_SUMMARY remain to be identified. In this study, we explored the change of liver ST:STUDY_SUMMARY metabolites in a diet-induced mouse NASH model. Identification of these ST:STUDY_SUMMARY metabolites will help to uncover the pathogenesis of NASH and explore the new ST:STUDY_SUMMARY therapeutic treatment. ST:INSTITUTE University of Missouri-Columbia ST:LAST_NAME Yang ST:FIRST_NAME Ming ST:ADDRESS University of Missouri ST:EMAIL yangmin@health.missouri.edu ST:PHONE 5738827141 #SUBJECT SU:SUBJECT_TYPE Mammal SU:SUBJECT_SPECIES Mus musculus SU:TAXONOMY_ID 10090 SU:GENDER Male #FACTORS #SUBJECT_SAMPLE_FACTORS: SUBJECT(optional)[tab]SAMPLE[tab]FACTORS(NAME:VALUE pairs separated by |)[tab]Raw file names and additional sample data SUBJECT_SAMPLE_FACTORS - C1 raw_data_time_format:High-fat diet SUBJECT_SAMPLE_FACTORS - C2 raw_data_time_format:High-fat diet SUBJECT_SAMPLE_FACTORS - C3 raw_data_time_format:High-fat diet SUBJECT_SAMPLE_FACTORS - C4 raw_data_time_format:High-fat diet SUBJECT_SAMPLE_FACTORS - C5 raw_data_time_format:High-fat diet SUBJECT_SAMPLE_FACTORS - C6 raw_data_time_format:High-fat diet SUBJECT_SAMPLE_FACTORS - C7 raw_data_time_format:High-fat diet SUBJECT_SAMPLE_FACTORS - C8 raw_data_time_format:High-fat diet SUBJECT_SAMPLE_FACTORS - C9 raw_data_time_format:High-fat diet SUBJECT_SAMPLE_FACTORS - C10 raw_data_time_format:High-fat diet SUBJECT_SAMPLE_FACTORS - A1 raw_data_time_format:High-fat diet + Antibiotic cocktail SUBJECT_SAMPLE_FACTORS - A2 raw_data_time_format:High-fat diet + Antibiotic cocktail SUBJECT_SAMPLE_FACTORS - A3 raw_data_time_format:High-fat diet + Antibiotic cocktail SUBJECT_SAMPLE_FACTORS - A4 raw_data_time_format:High-fat diet + Antibiotic cocktail SUBJECT_SAMPLE_FACTORS - A5 raw_data_time_format:High-fat diet + Antibiotic cocktail SUBJECT_SAMPLE_FACTORS - A6 raw_data_time_format:High-fat diet + Antibiotic cocktail SUBJECT_SAMPLE_FACTORS - A7 raw_data_time_format:High-fat diet + Antibiotic cocktail SUBJECT_SAMPLE_FACTORS - A8 raw_data_time_format:High-fat diet + Antibiotic cocktail SUBJECT_SAMPLE_FACTORS - A9 raw_data_time_format:High-fat diet + Antibiotic cocktail SUBJECT_SAMPLE_FACTORS - A10 raw_data_time_format:High-fat diet + Antibiotic cocktail SUBJECT_SAMPLE_FACTORS - N1 raw_data_time_format:ND SUBJECT_SAMPLE_FACTORS - N2 raw_data_time_format:ND SUBJECT_SAMPLE_FACTORS - N3 raw_data_time_format:ND SUBJECT_SAMPLE_FACTORS - N4 raw_data_time_format:ND SUBJECT_SAMPLE_FACTORS - N5 raw_data_time_format:ND SUBJECT_SAMPLE_FACTORS - N6 raw_data_time_format:ND SUBJECT_SAMPLE_FACTORS - N7 raw_data_time_format:ND SUBJECT_SAMPLE_FACTORS - N8 raw_data_time_format:ND SUBJECT_SAMPLE_FACTORS - N9 raw_data_time_format:ND SUBJECT_SAMPLE_FACTORS - N10 raw_data_time_format:ND #COLLECTION CO:COLLECTION_SUMMARY Liver biopsies from experimental mice were collected and immediately frozen in CO:COLLECTION_SUMMARY liquid nitrogen. CO:SAMPLE_TYPE Liver CO:STORAGE_CONDITIONS -80℃ #TREATMENT TR:TREATMENT_SUMMARY Mice were treated with a normal diet, high-fat high-sugar diet, or treated with TR:TREATMENT_SUMMARY a high-fat high-sugar diet and an antibiotic cocktail (ABX). #SAMPLEPREP SP:SAMPLEPREP_SUMMARY Briefly, liver tissues (50-100 mg) are typically homogenized in the extraction SP:SAMPLEPREP_SUMMARY solvent using a bead beater. These samples are then centrifuged at 13,000 g for SP:SAMPLEPREP_SUMMARY 15 min to pellet tissue and proteins. The supernatant is combined with water and SP:SAMPLEPREP_SUMMARY chloroform, including internal standards, to separate into the polar and SP:SAMPLEPREP_SUMMARY non-polar phases. SP:PROCESSING_STORAGE_CONDITIONS -80℃ SP:EXTRACT_STORAGE -80℃ #CHROMATOGRAPHY CH:CHROMATOGRAPHY_SUMMARY GC conditions included GC System (Agilent 6890) with column (DB-5MS, 60 m, 0.25 CH:CHROMATOGRAPHY_SUMMARY mm ID, 0.25 µm film thickness, J&W Scientific), and gas type helium. MS CH:CHROMATOGRAPHY_SUMMARY conditions included MS system (Agilent 5973 MSD) with electron ionization (EI CH:CHROMATOGRAPHY_SUMMARY ionization) with scan range from m/z 50 to m/z 650. CH:CHROMATOGRAPHY_TYPE GC CH:INSTRUMENT_NAME Agilent 6890N CH:COLUMN_NAME Agilent DB5-MS (30m x 0.25mm, 0.25um) #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:INSTRUMENT_NAME Agilent 5973 MS:INSTRUMENT_TYPE Ion trap MS:MS_TYPE EI MS:ION_MODE POSITIVE MS:MS_COMMENTS The data are deconvoluted using AMDIS and annotated using an in-house MS:MS_COMMENTS constructed spectral library MS:MS_RESULTS_FILE ST002156_AN003531_Results.txt UNITS:None Has m/z:Yes Has RT:Yes RT units:Minutes #END