{
"METABOLOMICS WORKBENCH":{"STUDY_ID":"ST001999","ANALYSIS_ID":"AN003261","VERSION":"1","CREATED_ON":"November 22, 2021, 12:33 pm"},

"PROJECT":{"PROJECT_TITLE":"Polyamine import and accumulation causes immunomodulation in macrophages engulfing apoptotic cells","PROJECT_SUMMARY":"Phagocytosis of apoptotic cells, termed efferocytosis, is critical for tissue homeostasis and drives anti-inflammatory programming in engulfing macrophages. Here, we assess metabolites in naïve and inflammatory macrophages following engulfment of multiple cellular and non-cellular targets. Efferocytosis leads to unique increases in the arginine-derived polyamines, spermidine and spermine, in vitro and in vivo. Surprisingly, polyamine accumulation after efferocytosis does not arise from retention of apoptotic cell metabolites or de novo synthesis, but from enhanced polyamine import that is dependent on Rac1, actin, and PI3 kinase. Blocking polyamine import prevents efferocytosis from suppressing macrophage IL-1beta or IL-6. This identifies efferocytosis as a trigger for polyamine import and accumulation, and imported polyamines as mediators of efferocytosis-induced immune reprogramming.","INSTITUTE":"University of Colorado Denver","LAST_NAME":"Haines","FIRST_NAME":"Julie","ADDRESS":"12801 E 17th Ave, Room 1303, Aurora, Colorado, 80045, USA","EMAIL":"julie.haines@cuanschutz.edu","PHONE":"3037243339"},

"STUDY":{"STUDY_TITLE":"Polyamine import and accumulation causes immunomodulation in macrophages engulfing apoptotic cells (Part 4)","STUDY_SUMMARY":"Phagocytosis of apoptotic cells, termed efferocytosis, is critical for tissue homeostasis and drives anti-inflammatory programming in engulfing macrophages. Here, we assess metabolites in naïve and inflammatory macrophages following engulfment of multiple cellular and non-cellular targets. Efferocytosis leads to unique increases in the arginine-derived polyamines, spermidine and spermine, in vitro and in vivo. Surprisingly, polyamine accumulation after efferocytosis does not arise from retention of apoptotic cell metabolites or de novo synthesis, but from enhanced polyamine import that is dependent on Rac1, actin, and PI3 kinase. Blocking polyamine import prevents efferocytosis from suppressing macrophage IL-1beta or IL-6. This identifies efferocytosis as a trigger for polyamine import and accumulation, and imported polyamines as mediators of efferocytosis-induced immune reprogramming.","INSTITUTE":"University of Colorado Denver","LAST_NAME":"Haines","FIRST_NAME":"Julie","ADDRESS":"12801 E 17th Ave, Room 1303, Aurora, Colorado, 80045, USA","EMAIL":"julie.haines@cuanschutz.edu","PHONE":"3037243339"},

"SUBJECT":{"SUBJECT_TYPE":"Cultured cells","SUBJECT_SPECIES":"Mus musculus","TAXONOMY_ID":"10090"},
"SUBJECT_SAMPLE_FACTORS":[
{
"Subject ID":"-",
"Sample ID":"ctrl 1",
"Factors":{"Category":"unexposed macrophages"},
"Additional sample data":{"RAW_FILE_NAME":"7+"}
},
{
"Subject ID":"-",
"Sample ID":"ctrl 2",
"Factors":{"Category":"unexposed macrophages"},
"Additional sample data":{"RAW_FILE_NAME":"8+"}
},
{
"Subject ID":"-",
"Sample ID":"ctrl 3",
"Factors":{"Category":"unexposed macrophages"},
"Additional sample data":{"RAW_FILE_NAME":"9+"}
},
{
"Subject ID":"-",
"Sample ID":"1 -AC 8h",
"Factors":{"Category":"non-engulfing"},
"Additional sample data":{"RAW_FILE_NAME":"10+"}
},
{
"Subject ID":"-",
"Sample ID":"2 -AC 8h",
"Factors":{"Category":"non-engulfing"},
"Additional sample data":{"RAW_FILE_NAME":"11+"}
},
{
"Subject ID":"-",
"Sample ID":"3 -AC 8h",
"Factors":{"Category":"non-engulfing"},
"Additional sample data":{"RAW_FILE_NAME":"12+"}
},
{
"Subject ID":"-",
"Sample ID":"1 +AC 8h",
"Factors":{"Category":"engulfing"},
"Additional sample data":{"RAW_FILE_NAME":"13+"}
},
{
"Subject ID":"-",
"Sample ID":"2 +AC 8h",
"Factors":{"Category":"engulfing"},
"Additional sample data":{"RAW_FILE_NAME":"14+"}
},
{
"Subject ID":"-",
"Sample ID":"3 +AC 8h",
"Factors":{"Category":"engulfing"},
"Additional sample data":{"RAW_FILE_NAME":"15+"}
}
],
"COLLECTION":{"COLLECTION_SUMMARY":"LPS-primed murine peritoneal macrophages were fed fluorescently-tagged apoptotic Jurkat cells for 1h. Unengulfed targets were washed off and macrophages were left to degrade targets for a further 7 hours. Macrophages were then collected and sorted into non-engulfing or engulfing populations, alongside control unexposed macrophages. Sorted cells were pelleted at 400xg and dry pellets were snap frozen and stored at -80C until metabolite extraction.","SAMPLE_TYPE":"Macrophages"},

"TREATMENT":{"TREATMENT_SUMMARY":"LPS-primed murine peritoneal macrophages were fed fluorescently-tagged apoptotic Jurkat cells for 1h. Unengulfed targets were washed off and macrophages were left to degrade targets for a further 7 hours. Macrophages were then collected and sorted into non-engulfing or engulfing populations, alongside control unexposed macrophages. Sorted cells were pelleted at 400xg and dry pellets were snap frozen and stored at -80C until metabolite extraction."},

"SAMPLEPREP":{"SAMPLEPREP_SUMMARY":"To process cells for assessment of intracellular metabolites, cells were pelleted at 400xg in tubes coated with 0.06% BSA. Supernatant was aspirated and discarded; residual liquid was carefully wicked away from the pellet with a kimwipe. Dry pellets were immediately snap frozen and stored at -80C until processing. To process culture supernatants for assessment of metabolites, supernatant was centrifuged at 400xg to pellet any cells. Cell-free supernatant was then transferred to a fresh tube, snap frozen, and stored at -80C until processing. Ultra-high pressure liquid chromatography-mass spectrometry (UHPLC-MS) was performed by the University of Colorado School of Medicine Metabolomics Core. Metabolites from frozen cell pellets were extracted at 2e6 cells/mL in ice cold 5:3:2 MeOH:acetonitrile:water (v/v/v). Media was thawed on ice and a 10 L aliquot treated with 240 L of the same extraction solution. Extractions were carried out using vigorous vortexing for 30 min at 4C. Supernatants were clarified by centrifugation (10 min, 18,000 g, 4C) and analyzed using a Thermo Vanquish UHPLC coupled to a Thermo Q Exactive mass spectrometer."},

"CHROMATOGRAPHY":{"CHROMATOGRAPHY_SUMMARY":"Isocratic flow at 250 uL/min using 95% phase A (0.1% formic acid in water) and 5% phase B (0.1% formic acid in acetonitrile).","CHROMATOGRAPHY_TYPE":"Reversed phase","INSTRUMENT_NAME":"Thermo Vanquish","COLUMN_NAME":"Phenomenex Kinetex C18 (150 x 2.1mm, 2.6 um)","FLOW_GRADIENT":"isocratic","FLOW_RATE":"250 ul/min","COLUMN_TEMPERATURE":"25"},

"ANALYSIS":{"ANALYSIS_TYPE":"MS"},

"MS":{"INSTRUMENT_NAME":"Thermo Q Exactive Orbitrap","INSTRUMENT_TYPE":"Orbitrap","MS_TYPE":"ESI","ION_MODE":"POSITIVE","MS_COMMENTS":"Global metabolomics analyses were performed using a 3 min isocratic run in positive and negative ion modes (separate runs) as described previously (Nemkov et al., 2015, Nemkov et al., 2017); stable isotope tracing samples were analyzed using a 5 min C18 gradient in positive and negative ion modes (separate runs) as described (Nemkov et al., 2019, Gehrke et al., 2019). For all analyses, the MS scanned in MS1 mode across the m/z range of 65 to 950. Peaks were annotated (in conjunction with the KEGG database), integrated, and quality control performed using Maven (Princeton University) as described. Stable isotope tracing results were isotopically corrected for the natural abundance of 13C1, 13C2, 15N1, and 15N2 (Nemkov et al., 2017)."},

"MS_METABOLITE_DATA":{
"Units":"peak area",

"Data":[{"Metabolite":"L-alanine","ctrl 1":"6.70E+05","ctrl 2":"6.41E+05","ctrl 3":"6.96E+05","1 -AC 8h":"6.47E+05","2 -AC 8h":"7.43E+05","3 -AC 8h":"7.44E+05","1 +AC 8h":"6.68E+05","2 +AC 8h":"7.25E+05","3 +AC 8h":"6.57E+05"},{"Metabolite":"L-arginine","ctrl 1":"3.03E+05","ctrl 2":"2.99E+05","ctrl 3":"4.56E+05","1 -AC 8h":"1.70E+05","2 -AC 8h":"1.77E+05","3 -AC 8h":"1.49E+05","1 +AC 8h":"1.97E+06","2 +AC 8h":"1.36E+06","3 +AC 8h":"2.74E+06"},{"Metabolite":"L-asparagine","ctrl 1":"1.94E+04","ctrl 2":"3.90E+04","ctrl 3":"2.19E+04","1 -AC 8h":"0.00E+00","2 -AC 8h":"1.56E+04","3 -AC 8h":"1.68E+04","1 +AC 8h":"1.41E+05","2 +AC 8h":"8.35E+04","3 +AC 8h":"9.89E+04"},{"Metabolite":"L-aspartate","ctrl 1":"1.00E+05","ctrl 2":"8.02E+04","ctrl 3":"9.67E+04","1 -AC 8h":"1.53E+04","2 -AC 8h":"2.82E+04","3 -AC 8h":"2.65E+04","1 +AC 8h":"2.36E+05","2 +AC 8h":"1.52E+05","3 +AC 8h":"1.52E+05"},{"Metabolite":"L-cysteine","ctrl 1":"1.38E+05","ctrl 2":"1.39E+05","ctrl 3":"9.36E+04","1 -AC 8h":"1.18E+04","2 -AC 8h":"4.57E+04","3 -AC 8h":"1.33E+04","1 +AC 8h":"4.54E+05","2 +AC 8h":"4.20E+05","3 +AC 8h":"3.08E+05"},{"Metabolite":"L-glutamate","ctrl 1":"8.71E+05","ctrl 2":"7.34E+05","ctrl 3":"6.42E+05","1 -AC 8h":"2.17E+05","2 -AC 8h":"3.23E+05","3 -AC 8h":"2.45E+05","1 +AC 8h":"1.48E+06","2 +AC 8h":"9.97E+05","3 +AC 8h":"9.94E+05"},{"Metabolite":"L-glutamine","ctrl 1":"3.45E+05","ctrl 2":"3.62E+05","ctrl 3":"4.89E+05","1 -AC 8h":"1.72E+05","2 -AC 8h":"1.63E+05","3 -AC 8h":"1.23E+05","1 +AC 8h":"1.43E+06","2 +AC 8h":"1.07E+06","3 +AC 8h":"1.12E+06"},{"Metabolite":"L-histidine","ctrl 1":"7.33E+04","ctrl 2":"9.43E+04","ctrl 3":"1.33E+05","1 -AC 8h":"5.97E+04","2 -AC 8h":"6.12E+04","3 -AC 8h":"4.95E+04","1 +AC 8h":"4.49E+05","2 +AC 8h":"2.71E+05","3 +AC 8h":"3.39E+05"},{"Metabolite":"L-leucine","ctrl 1":"6.89E+05","ctrl 2":"9.60E+05","ctrl 3":"1.37E+06","1 -AC 8h":"5.49E+05","2 -AC 8h":"5.18E+05","3 -AC 8h":"6.71E+05","1 +AC 8h":"3.37E+06","2 +AC 8h":"2.68E+06","3 +AC 8h":"2.58E+06"},{"Metabolite":"L-lysine","ctrl 1":"1.86E+05","ctrl 2":"2.34E+05","ctrl 3":"4.05E+05","1 -AC 8h":"1.90E+05","2 -AC 8h":"1.95E+05","3 -AC 8h":"1.99E+05","1 +AC 8h":"3.11E+06","2 +AC 8h":"2.18E+06","3 +AC 8h":"2.52E+06"},{"Metabolite":"L-methionine","ctrl 1":"2.53E+04","ctrl 2":"2.46E+04","ctrl 3":"8.23E+03","1 -AC 8h":"0.00E+00","2 -AC 8h":"0.00E+00","3 -AC 8h":"0.00E+00","1 +AC 8h":"1.37E+05","2 +AC 8h":"8.78E+04","3 +AC 8h":"7.67E+04"},{"Metabolite":"L-phenylalanine","ctrl 1":"1.46E+05","ctrl 2":"1.91E+05","ctrl 3":"4.47E+05","1 -AC 8h":"2.67E+05","2 -AC 8h":"1.79E+05","3 -AC 8h":"2.45E+05","1 +AC 8h":"9.89E+05","2 +AC 8h":"7.37E+05","3 +AC 8h":"6.06E+05"},{"Metabolite":"L-proline","ctrl 1":"1.53E+06","ctrl 2":"2.81E+06","ctrl 3":"2.04E+06","1 -AC 8h":"1.47E+06","2 -AC 8h":"1.46E+06","3 -AC 8h":"1.70E+06","1 +AC 8h":"6.14E+06","2 +AC 8h":"4.90E+06","3 +AC 8h":"4.06E+06"},{"Metabolite":"L-serine","ctrl 1":"3.62E+04","ctrl 2":"6.12E+04","ctrl 3":"5.43E+04","1 -AC 8h":"2.54E+04","2 -AC 8h":"2.38E+04","3 -AC 8h":"1.23E+04","1 +AC 8h":"1.64E+05","2 +AC 8h":"8.43E+04","3 +AC 8h":"1.10E+05"},{"Metabolite":"L-threonine","ctrl 1":"1.03E+05","ctrl 2":"1.04E+05","ctrl 3":"9.32E+04","1 -AC 8h":"3.81E+04","2 -AC 8h":"5.73E+04","3 -AC 8h":"1.76E+04","1 +AC 8h":"4.88E+05","2 +AC 8h":"4.28E+05","3 +AC 8h":"3.41E+05"},{"Metabolite":"L-tryptophan","ctrl 1":"4.92E+05","ctrl 2":"5.83E+05","ctrl 3":"6.33E+05","1 -AC 8h":"7.15E+05","2 -AC 8h":"4.52E+05","3 -AC 8h":"7.64E+05","1 +AC 8h":"4.58E+06","2 +AC 8h":"3.86E+06","3 +AC 8h":"3.46E+06"},{"Metabolite":"L-tyrosine","ctrl 1":"2.05E+05","ctrl 2":"2.08E+05","ctrl 3":"2.61E+05","1 -AC 8h":"1.32E+05","2 -AC 8h":"1.84E+05","3 -AC 8h":"1.99E+05","1 +AC 8h":"4.75E+05","2 +AC 8h":"4.63E+05","3 +AC 8h":"3.60E+05"},{"Metabolite":"L-valine","ctrl 1":"2.94E+06","ctrl 2":"3.56E+06","ctrl 3":"3.03E+06","1 -AC 8h":"2.27E+06","2 -AC 8h":"2.61E+06","3 -AC 8h":"2.53E+06","1 +AC 8h":"4.00E+06","2 +AC 8h":"3.73E+06","3 +AC 8h":"2.74E+06"},{"Metabolite":"AMP","ctrl 1":"1.62E+05","ctrl 2":"1.90E+05","ctrl 3":"1.07E+05","1 -AC 8h":"5.40E+04","2 -AC 8h":"8.75E+04","3 -AC 8h":"6.60E+04","1 +AC 8h":"3.45E+05","2 +AC 8h":"1.98E+05","3 +AC 8h":"2.17E+05"},{"Metabolite":"Adenosine","ctrl 1":"3.45E+05","ctrl 2":"6.07E+05","ctrl 3":"3.83E+05","1 -AC 8h":"1.39E+05","2 -AC 8h":"1.51E+05","3 -AC 8h":"1.23E+05","1 +AC 8h":"3.15E+05","2 +AC 8h":"2.38E+05","3 +AC 8h":"7.77E+04"},{"Metabolite":"Cytosine","ctrl 1":"4.75E+04","ctrl 2":"1.18E+05","ctrl 3":"4.03E+04","1 -AC 8h":"1.96E+05","2 -AC 8h":"2.61E+05","3 -AC 8h":"3.34E+05","1 +AC 8h":"5.44E+06","2 +AC 8h":"5.53E+06","3 +AC 8h":"3.21E+06"},{"Metabolite":"IMP","ctrl 1":"1.13E+05","ctrl 2":"1.13E+05","ctrl 3":"5.93E+04","1 -AC 8h":"1.72E+04","2 -AC 8h":"2.06E+04","3 -AC 8h":"1.37E+04","1 +AC 8h":"1.12E+05","2 +AC 8h":"3.54E+04","3 +AC 8h":"5.01E+04"},{"Metabolite":"Hypoxanthine","ctrl 1":"3.12E+05","ctrl 2":"6.46E+05","ctrl 3":"2.76E+05","1 -AC 8h":"3.30E+05","2 -AC 8h":"3.12E+05","3 -AC 8h":"3.44E+05","1 +AC 8h":"3.39E+06","2 +AC 8h":"2.57E+06","3 +AC 8h":"1.40E+06"},{"Metabolite":"Urate","ctrl 1":"3.62E+04","ctrl 2":"3.72E+04","ctrl 3":"2.15E+04","1 -AC 8h":"9.78E+03","2 -AC 8h":"9.58E+03","3 -AC 8h":"1.55E+04","1 +AC 8h":"6.90E+04","2 +AC 8h":"3.91E+04","3 +AC 8h":"3.51E+04"},{"Metabolite":"Nicotinamide","ctrl 1":"3.18E+06","ctrl 2":"8.07E+06","ctrl 3":"3.38E+06","1 -AC 8h":"1.77E+06","2 -AC 8h":"2.50E+06","3 -AC 8h":"2.41E+06","1 +AC 8h":"7.33E+06","2 +AC 8h":"5.96E+06","3 +AC 8h":"1.86E+06"},{"Metabolite":"Thymine","ctrl 1":"5.87E+04","ctrl 2":"7.70E+04","ctrl 3":"6.32E+04","1 -AC 8h":"6.24E+04","2 -AC 8h":"6.03E+04","3 -AC 8h":"7.24E+04","1 +AC 8h":"1.30E+05","2 +AC 8h":"1.23E+05","3 +AC 8h":"1.56E+05"},{"Metabolite":"Glutathione","ctrl 1":"5.49E+05","ctrl 2":"6.56E+05","ctrl 3":"4.33E+05","1 -AC 8h":"3.57E+05","2 -AC 8h":"2.73E+05","3 -AC 8h":"2.02E+05","1 +AC 8h":"1.20E+06","2 +AC 8h":"9.13E+05","3 +AC 8h":"3.41E+05"},{"Metabolite":"S-Adenosyl-L-methionine","ctrl 1":"2.07E+04","ctrl 2":"4.78E+04","ctrl 3":"3.14E+04","1 -AC 8h":"0.00E+00","2 -AC 8h":"2.55E+04","3 -AC 8h":"2.65E+04","1 +AC 8h":"1.61E+05","2 +AC 8h":"9.08E+04","3 +AC 8h":"5.93E+04"},{"Metabolite":"Ornithine","ctrl 1":"8.35E+04","ctrl 2":"1.06E+05","ctrl 3":"1.32E+05","1 -AC 8h":"7.87E+04","2 -AC 8h":"8.52E+04","3 -AC 8h":"7.19E+04","1 +AC 8h":"5.57E+05","2 +AC 8h":"3.42E+05","3 +AC 8h":"3.88E+05"},{"Metabolite":"L-Citrulline","ctrl 1":"4.25E+04","ctrl 2":"7.92E+04","ctrl 3":"5.26E+04","1 -AC 8h":"4.63E+04","2 -AC 8h":"3.54E+04","3 -AC 8h":"1.98E+04","1 +AC 8h":"8.00E+04","2 +AC 8h":"5.57E+04","3 +AC 8h":"5.42E+04"},{"Metabolite":"Cadaverine","ctrl 1":"1.84E+04","ctrl 2":"1.63E+05","ctrl 3":"5.57E+04","1 -AC 8h":"3.47E+04","2 -AC 8h":"1.50E+05","3 -AC 8h":"5.10E+04","1 +AC 8h":"6.80E+04","2 +AC 8h":"4.08E+05","3 +AC 8h":"2.60E+04"},{"Metabolite":"Spermidine","ctrl 1":"4.14E+06","ctrl 2":"8.39E+06","ctrl 3":"4.10E+06","1 -AC 8h":"1.31E+07","2 -AC 8h":"8.38E+06","3 -AC 8h":"1.07E+07","1 +AC 8h":"2.83E+07","2 +AC 8h":"1.95E+07","3 +AC 8h":"1.02E+07"},{"Metabolite":"Spermine","ctrl 1":"2.47E+05","ctrl 2":"4.74E+05","ctrl 3":"2.27E+05","1 -AC 8h":"1.51E+06","2 -AC 8h":"7.41E+05","3 -AC 8h":"9.70E+05","1 +AC 8h":"6.90E+06","2 +AC 8h":"5.06E+06","3 +AC 8h":"1.64E+06"},{"Metabolite":"Putrescine","ctrl 1":"1.52E+05","ctrl 2":"2.03E+05","ctrl 3":"1.56E+05","1 -AC 8h":"1.04E+05","2 -AC 8h":"1.22E+05","3 -AC 8h":"1.43E+05","1 +AC 8h":"3.31E+05","2 +AC 8h":"2.56E+05","3 +AC 8h":"2.54E+05"},{"Metabolite":"D-Glucosamine","ctrl 1":"7.00E+04","ctrl 2":"2.13E+05","ctrl 3":"8.22E+04","1 -AC 8h":"1.26E+05","2 -AC 8h":"1.65E+05","3 -AC 8h":"1.76E+05","1 +AC 8h":"1.37E+05","2 +AC 8h":"1.96E+05","3 +AC 8h":"1.05E+05"},{"Metabolite":"Creatine","ctrl 1":"1.19E+07","ctrl 2":"2.22E+07","ctrl 3":"9.17E+06","1 -AC 8h":"6.81E+06","2 -AC 8h":"7.36E+06","3 -AC 8h":"7.10E+06","1 +AC 8h":"1.52E+07","2 +AC 8h":"1.08E+07","3 +AC 8h":"7.36E+06"},{"Metabolite":"Creatinine","ctrl 1":"1.18E+05","ctrl 2":"1.88E+05","ctrl 3":"2.55E+05","1 -AC 8h":"8.45E+04","2 -AC 8h":"1.08E+05","3 -AC 8h":"1.41E+05","1 +AC 8h":"2.68E+05","2 +AC 8h":"2.15E+05","3 +AC 8h":"2.23E+05"},{"Metabolite":"N-Acetylornithine","ctrl 1":"1.43E+04","ctrl 2":"4.42E+04","ctrl 3":"2.40E+04","1 -AC 8h":"1.27E+04","2 -AC 8h":"1.20E+04","3 -AC 8h":"2.80E+04","1 +AC 8h":"1.08E+05","2 +AC 8h":"9.26E+04","3 +AC 8h":"5.94E+04"},{"Metabolite":"Taurine","ctrl 1":"2.33E+05","ctrl 2":"2.17E+05","ctrl 3":"1.39E+05","1 -AC 8h":"6.31E+04","2 -AC 8h":"5.60E+04","3 -AC 8h":"6.80E+04","1 +AC 8h":"1.22E+05","2 +AC 8h":"5.85E+04","3 +AC 8h":"8.50E+04"},{"Metabolite":"L-Methionine S-oxide","ctrl 1":"4.07E+04","ctrl 2":"4.49E+04","ctrl 3":"5.42E+04","1 -AC 8h":"5.99E+04","2 -AC 8h":"4.16E+04","3 -AC 8h":"5.13E+04","1 +AC 8h":"6.02E+05","2 +AC 8h":"4.97E+05","3 +AC 8h":"2.84E+05"},{"Metabolite":"Indole","ctrl 1":"4.57E+05","ctrl 2":"3.97E+05","ctrl 3":"5.57E+05","1 -AC 8h":"5.59E+05","2 -AC 8h":"4.30E+05","3 -AC 8h":"3.99E+05","1 +AC 8h":"5.81E+05","2 +AC 8h":"5.28E+05","3 +AC 8h":"4.97E+05"},{"Metabolite":"Indole-3-acetate","ctrl 1":"1.64E+04","ctrl 2":"1.88E+04","ctrl 3":"5.45E+04","1 -AC 8h":"1.62E+04","2 -AC 8h":"1.52E+04","3 -AC 8h":"6.04E+03","1 +AC 8h":"9.60E+04","2 +AC 8h":"4.44E+04","3 +AC 8h":"2.92E+04"},{"Metabolite":"Sphingosine","ctrl 1":"1.98E+05","ctrl 2":"7.88E+04","ctrl 3":"2.49E+05","1 -AC 8h":"2.79E+05","2 -AC 8h":"2.16E+05","3 -AC 8h":"9.89E+04","1 +AC 8h":"2.17E+05","2 +AC 8h":"9.60E+04","3 +AC 8h":"2.16E+05"},{"Metabolite":"Acetylcholine","ctrl 1":"8.06E+05","ctrl 2":"1.33E+06","ctrl 3":"8.26E+05","1 -AC 8h":"4.36E+05","2 -AC 8h":"5.08E+05","3 -AC 8h":"5.25E+05","1 +AC 8h":"1.57E+06","2 +AC 8h":"1.43E+06","3 +AC 8h":"9.80E+05"},{"Metabolite":"Choline","ctrl 1":"2.10E+07","ctrl 2":"3.24E+07","ctrl 3":"2.13E+07","1 -AC 8h":"1.56E+07","2 -AC 8h":"1.30E+07","3 -AC 8h":"1.09E+07","1 +AC 8h":"5.07E+07","2 +AC 8h":"3.32E+07","3 +AC 8h":"2.66E+07"},{"Metabolite":"L-Carnitine","ctrl 1":"1.34E+06","ctrl 2":"1.73E+06","ctrl 3":"1.01E+06","1 -AC 8h":"1.29E+06","2 -AC 8h":"1.41E+06","3 -AC 8h":"1.28E+06","1 +AC 8h":"2.78E+06","2 +AC 8h":"2.47E+06","3 +AC 8h":"1.80E+06"},{"Metabolite":"acetyl-carnitine (acyl-C2)","ctrl 1":"5.16E+06","ctrl 2":"8.76E+06","ctrl 3":"4.84E+06","1 -AC 8h":"2.49E+06","2 -AC 8h":"2.51E+06","3 -AC 8h":"2.34E+06","1 +AC 8h":"5.04E+06","2 +AC 8h":"4.82E+06","3 +AC 8h":"2.16E+06"},{"Metabolite":"propionyl-carnitine (acyl-C3)","ctrl 1":"5.31E+04","ctrl 2":"1.01E+05","ctrl 3":"4.22E+04","1 -AC 8h":"5.35E+04","2 -AC 8h":"4.83E+04","3 -AC 8h":"5.64E+04","1 +AC 8h":"1.90E+05","2 +AC 8h":"1.32E+05","3 +AC 8h":"4.92E+04"},{"Metabolite":"butanoyl-l-carnitine (acyl-C4)","ctrl 1":"2.22E+04","ctrl 2":"1.78E+04","ctrl 3":"1.74E+04","1 -AC 8h":"6.15E+04","2 -AC 8h":"2.37E+04","3 -AC 8h":"2.55E+04","1 +AC 8h":"2.57E+04","2 +AC 8h":"2.68E+04","3 +AC 8h":"2.31E+04"},{"Metabolite":"acyl-C4-OH (Hydroxybutyrylcarnitine)","ctrl 1":"1.60E+05","ctrl 2":"2.68E+05","ctrl 3":"1.40E+05","1 -AC 8h":"4.96E+04","2 -AC 8h":"6.42E+04","3 -AC 8h":"8.26E+04","1 +AC 8h":"1.10E+05","2 +AC 8h":"1.24E+05","3 +AC 8h":"8.33E+04"},{"Metabolite":"Propionic/Propanoic acid","ctrl 1":"3.46E+05","ctrl 2":"2.69E+05","ctrl 3":"2.78E+05","1 -AC 8h":"3.17E+05","2 -AC 8h":"3.00E+05","3 -AC 8h":"2.57E+05","1 +AC 8h":"2.56E+05","2 +AC 8h":"2.60E+05","3 +AC 8h":"3.22E+05"},{"Metabolite":"Methylenediurea","ctrl 1":"8.09E+04","ctrl 2":"1.98E+05","ctrl 3":"6.80E+04","1 -AC 8h":"6.26E+04","2 -AC 8h":"5.54E+04","3 -AC 8h":"5.65E+04","1 +AC 8h":"1.19E+05","2 +AC 8h":"9.84E+04","3 +AC 8h":"5.29E+04"},{"Metabolite":"2-Deoxy-alpha-D-glucoside","ctrl 1":"6.66E+05","ctrl 2":"5.39E+05","ctrl 3":"5.93E+05","1 -AC 8h":"5.40E+05","2 -AC 8h":"4.77E+05","3 -AC 8h":"4.01E+05","1 +AC 8h":"9.63E+05","2 +AC 8h":"5.41E+05","3 +AC 8h":"6.23E+05"},{"Metabolite":"N-Acyl-D-mannosaminolactone","ctrl 1":"9.41E+05","ctrl 2":"1.16E+06","ctrl 3":"1.08E+06","1 -AC 8h":"1.65E+06","2 -AC 8h":"1.68E+06","3 -AC 8h":"1.84E+06","1 +AC 8h":"1.37E+06","2 +AC 8h":"1.63E+06","3 +AC 8h":"1.10E+06"},{"Metabolite":"9-Oxononanoic acid","ctrl 1":"5.03E+05","ctrl 2":"6.29E+05","ctrl 3":"5.13E+05","1 -AC 8h":"4.68E+05","2 -AC 8h":"4.60E+05","3 -AC 8h":"4.84E+05","1 +AC 8h":"5.22E+05","2 +AC 8h":"6.10E+05","3 +AC 8h":"5.49E+05"},{"Metabolite":"2-Oxo-7-methylthioheptanoic acid","ctrl 1":"2.47E+04","ctrl 2":"2.46E+04","ctrl 3":"5.81E+04","1 -AC 8h":"3.34E+04","2 -AC 8h":"1.80E+04","3 -AC 8h":"1.98E+04","1 +AC 8h":"7.68E+05","2 +AC 8h":"5.78E+05","3 +AC 8h":"5.59E+05"}],

"Metabolites":[{"Metabolite":"L-alanine","KEGG ID":"C00041","m/z":"90.0555","r.t.":"0.93"},{"Metabolite":"L-arginine","KEGG ID":"C00062","m/z":"175.1188","r.t.":"0.63"},{"Metabolite":"L-asparagine","KEGG ID":"C00152","m/z":"133.0607","r.t.":"0.66"},{"Metabolite":"L-aspartate","KEGG ID":"C00049","m/z":"134.0448","r.t.":"0.66"},{"Metabolite":"L-cysteine","KEGG ID":"C00097","m/z":"122.0272","r.t.":"0.67"},{"Metabolite":"L-glutamate","KEGG ID":"C00025","m/z":"148.0603","r.t.":"0.65"},{"Metabolite":"L-glutamine","KEGG ID":"C00064","m/z":"147.0763","r.t.":"0.65"},{"Metabolite":"L-histidine","KEGG ID":"C00135","m/z":"156.0766","r.t.":"0.62"},{"Metabolite":"L-leucine","KEGG ID":"C00123","m/z":"132.1019","r.t.":"0.68"},{"Metabolite":"L-lysine","KEGG ID":"C00047","m/z":"147.1126","r.t.":"0.60"},{"Metabolite":"L-methionine","KEGG ID":"C00073","m/z":"150.0583","r.t.":"0.67"},{"Metabolite":"L-phenylalanine","KEGG ID":"C00079","m/z":"166.0861","r.t.":"1.44"},{"Metabolite":"L-proline","KEGG ID":"C00148","m/z":"116.0708","r.t.":"0.67"},{"Metabolite":"L-serine","KEGG ID":"C00065","m/z":"106.0502","r.t.":"0.68"},{"Metabolite":"L-threonine","KEGG ID":"C00188","m/z":"120.0657","r.t.":"0.65"},{"Metabolite":"L-tryptophan","KEGG ID":"C00078","m/z":"205.0969","r.t.":"1.72"},{"Metabolite":"L-tyrosine","KEGG ID":"C00082","m/z":"182.0797","r.t.":"0.65"},{"Metabolite":"L-valine","KEGG ID":"C00183","m/z":"118.0864","r.t.":"0.66"},{"Metabolite":"AMP","KEGG ID":"C00020","m/z":"348.0693","r.t.":"0.68"},{"Metabolite":"Adenosine","KEGG ID":"C00212","m/z":"268.1031","r.t.":"0.72"},{"Metabolite":"Cytosine","KEGG ID":"C00380","m/z":"112.0508","r.t.":"0.78"},{"Metabolite":"IMP","KEGG ID":"C00130","m/z":"349.0533","r.t.":"0.78"},{"Metabolite":"Hypoxanthine","KEGG ID":"C00262","m/z":"137.0457","r.t.":"0.73"},{"Metabolite":"Urate","KEGG ID":"C00366","m/z":"169.0354","r.t.":"0.74"},{"Metabolite":"Nicotinamide","KEGG ID":"C00153","m/z":"123.0554","r.t.":"0.76"},{"Metabolite":"Thymine","KEGG ID":"C00178","m/z":"127.0503","r.t.":"0.67"},{"Metabolite":"Glutathione","KEGG ID":"C00051","m/z":"308.0902","r.t.":"0.83"},{"Metabolite":"S-Adenosyl-L-methionine","KEGG ID":"C00019","m/z":"399.1433","r.t.":"0.65"},{"Metabolite":"Ornithine","KEGG ID":"C01602","m/z":"133.0971","r.t.":"0.60"},{"Metabolite":"L-Citrulline","KEGG ID":"C00327","m/z":"176.1027","r.t.":"0.65"},{"Metabolite":"Cadaverine","KEGG ID":"C01672","m/z":"103.1233","r.t.":"0.60"},{"Metabolite":"Spermidine","KEGG ID":"C00315","m/z":"146.1650","r.t.":"0.61"},{"Metabolite":"Spermine","KEGG ID":"C00750","m/z":"203.2227","r.t.":"0.60"},{"Metabolite":"Putrescine","KEGG ID":"C00134","m/z":"89.1079","r.t.":"0.61"},{"Metabolite":"D-Glucosamine","KEGG ID":"C00329","m/z":"180.0864","r.t.":"0.72"},{"Metabolite":"Creatine","KEGG ID":"C00300","m/z":"132.0767","r.t.":"0.68"},{"Metabolite":"Creatinine","KEGG ID":"C00791","m/z":"114.0664","r.t.":"0.66"},{"Metabolite":"N-Acetylornithine","KEGG ID":"C00437","m/z":"175.1075","r.t.":"0.67"},{"Metabolite":"Taurine","KEGG ID":"C00245","m/z":"126.0220","r.t.":"0.70"},{"Metabolite":"L-Methionine S-oxide","KEGG ID":"C02989","m/z":"166.0531","r.t.":"0.85"},{"Metabolite":"Indole","KEGG ID":"C00463","m/z":"118.0653","r.t.":"1.89"},{"Metabolite":"Indole-3-acetate","KEGG ID":"C00954","m/z":"176.0703","r.t.":"1.90"},{"Metabolite":"Sphingosine","KEGG ID":"C00319","m/z":"300.2889","r.t.":"1.92"},{"Metabolite":"Acetylcholine","KEGG ID":"C01996","m/z":"146.1174","r.t.":"0.68"},{"Metabolite":"Choline","KEGG ID":"C00114","m/z":"104.1074","r.t.":"0.65"},{"Metabolite":"L-Carnitine","KEGG ID":"C00318","m/z":"162.1123","r.t.":"0.65"},{"Metabolite":"acetyl-carnitine (acyl-C2)","KEGG ID":"C02571","m/z":"204.1227","r.t.":"0.67"},{"Metabolite":"propionyl-carnitine (acyl-C3)","KEGG ID":"C03017","m/z":"218.1383","r.t.":"0.68"},{"Metabolite":"butanoyl-l-carnitine (acyl-C4)","KEGG ID":"C02862","m/z":"232.1538","r.t.":"1.60"},{"Metabolite":"acyl-C4-OH (Hydroxybutyrylcarnitine)","KEGG ID":"HMDB13127","m/z":"248.1486","r.t.":"0.67"},{"Metabolite":"Propionic/Propanoic acid","KEGG ID":"C00163","m/z":"75.0448","r.t.":"1.69"},{"Metabolite":"Methylenediurea","KEGG ID":"C06381","m/z":"133.0735","r.t.":"0.66"},{"Metabolite":"2-Deoxy-alpha-D-glucoside","KEGG ID":"C03573","m/z":"164.0679","r.t.":"0.58"},{"Metabolite":"N-Acyl-D-mannosaminolactone","KEGG ID":"C03792","m/z":"205.0564","r.t.":"0.65"},{"Metabolite":"9-Oxononanoic acid","KEGG ID":"C16322","m/z":"173.1170","r.t.":"1.81"},{"Metabolite":"2-Oxo-7-methylthioheptanoic acid","KEGG ID":"C17220","m/z":"191.0760","r.t.":"0.62"}]
}

}