{
"METABOLOMICS WORKBENCH":{"STUDY_ID":"ST002729","ANALYSIS_ID":"AN004424","VERSION":"1","CREATED_ON":"11-09-2023"},

"PROJECT":{"PROJECT_TITLE":"Role of AICARP, an endogenous AMPK activator, in improved endurance capacity in diabetic mice during SGLT2 inhibition.","PROJECT_TYPE":"Wide targeted metabolomics & lipidomics","PROJECT_SUMMARY":"Diabetes is often associated with increased risk of deleterious muscle mass and function or sarcopenia, thus leading to physical inactivity and metabolic disorders. Sodium-glucose cotransporter 2 (SGLT2) inhibitors are an oral antidiabetic drug that promotes urinary excretion of glucose in the renal proximal tubules. However, how SGLT2 inhibition affects the skeletal muscle function in patients with diabetes remains elusive. Here, we examined the differential effect of CANA on the oxidative soleus and glycolytic EDL muscles from genetically obese diabetic db/db mice.","INSTITUTE":"Medical Institute of Bioregulation, Kyushu University","DEPARTMENT":"Metabolomics","LABORATORY":"Bamba lab.","LAST_NAME":"Takahashi","FIRST_NAME":"Masatomo","ADDRESS":"Maidashi 3-1-1, Higashi-ku, Fukuoka, Fukuoka, 8128582, Japan","EMAIL":"m-takahashi@bioreg.kyushu-u.ac.jp","PHONE":"0926426171","PUBLICATIONS":"https://onlinelibrary.wiley.com/doi/full/10.1002/jcsm.13350","DOI":"http://dx.doi.org/10.21228/M8VT66"},

"STUDY":{"STUDY_TITLE":"Improved Endurance Capacity of Diabetic Mice during SGLT2 Inhibition: Potential Role of AICARP, an Endogenous AMPK Activator.","STUDY_SUMMARY":"Diabetes is associated with an increased risk of deleterious changes in muscle mass and function or sarcopenia, leading to physical inactivity and worsening glycemic control. Given the negative energy balance during sodium-glucose cotransporter 2 (SGLT2) inhibition, whether SGLT2 inhibitors affect skeletal muscle mass and function is a matter of concern. However, how SGLT2 inhibition affects the skeletal muscle function in patients with diabetes remains insufficiently explored. We aimed to explore the effects of canagliflozin (CANA), an SGLT2 inhibitor, on skeletal muscles in genetically diabetic db/db mice.","INSTITUTE":"Medical Institute of Bioregulation, Kyushu University","LAST_NAME":"Takahashi","FIRST_NAME":"Masatomo","ADDRESS":"Maidashi 3-1-1, Higashi-ku, Fukuoka, Fukuoka, 8128582, Japan","EMAIL":"m-takahashi@bioreg.kyushu-u.ac.jp","PHONE":"0926426171","SUBMIT_DATE":"2023-05-25"},

"SUBJECT":{"SUBJECT_TYPE":"Mammal","SUBJECT_SPECIES":"Mus musculus","TAXONOMY_ID":"10090"},
"SUBJECT_SAMPLE_FACTORS":[
{
"Subject ID":"-",
"Sample ID":"13",
"Factors":{"Animal":"db/db","Treatment":"Cana"},
"Additional sample data":{"RAW_FILE_NAME":"13"}
},
{
"Subject ID":"-",
"Sample ID":"14",
"Factors":{"Animal":"db/db","Treatment":"Cana"},
"Additional sample data":{"RAW_FILE_NAME":"14"}
},
{
"Subject ID":"-",
"Sample ID":"15",
"Factors":{"Animal":"db/db","Treatment":"Cana"},
"Additional sample data":{"RAW_FILE_NAME":"15"}
},
{
"Subject ID":"-",
"Sample ID":"16",
"Factors":{"Animal":"db/db","Treatment":"Cana"},
"Additional sample data":{"RAW_FILE_NAME":"16"}
},
{
"Subject ID":"-",
"Sample ID":"61",
"Factors":{"Animal":"db/db","Treatment":"Cana"},
"Additional sample data":{"RAW_FILE_NAME":"61"}
},
{
"Subject ID":"-",
"Sample ID":"62",
"Factors":{"Animal":"db/db","Treatment":"Cana"},
"Additional sample data":{"RAW_FILE_NAME":"62"}
},
{
"Subject ID":"-",
"Sample ID":"63",
"Factors":{"Animal":"db/db","Treatment":"Cana"},
"Additional sample data":{"RAW_FILE_NAME":"63"}
},
{
"Subject ID":"-",
"Sample ID":"64",
"Factors":{"Animal":"db/db","Treatment":"Cana"},
"Additional sample data":{"RAW_FILE_NAME":"64"}
},
{
"Subject ID":"-",
"Sample ID":"10",
"Factors":{"Animal":"db/db","Treatment":"vehicle"},
"Additional sample data":{"RAW_FILE_NAME":"10"}
},
{
"Subject ID":"-",
"Sample ID":"11",
"Factors":{"Animal":"db/db","Treatment":"vehicle"},
"Additional sample data":{"RAW_FILE_NAME":"11"}
},
{
"Subject ID":"-",
"Sample ID":"12",
"Factors":{"Animal":"db/db","Treatment":"vehicle"},
"Additional sample data":{"RAW_FILE_NAME":"12"}
},
{
"Subject ID":"-",
"Sample ID":"57",
"Factors":{"Animal":"db/db","Treatment":"vehicle"},
"Additional sample data":{"RAW_FILE_NAME":"57"}
},
{
"Subject ID":"-",
"Sample ID":"58",
"Factors":{"Animal":"db/db","Treatment":"vehicle"},
"Additional sample data":{"RAW_FILE_NAME":"58"}
},
{
"Subject ID":"-",
"Sample ID":"59",
"Factors":{"Animal":"db/db","Treatment":"vehicle"},
"Additional sample data":{"RAW_FILE_NAME":"59"}
},
{
"Subject ID":"-",
"Sample ID":"60",
"Factors":{"Animal":"db/db","Treatment":"vehicle"},
"Additional sample data":{"RAW_FILE_NAME":"60"}
},
{
"Subject ID":"-",
"Sample ID":"9",
"Factors":{"Animal":"db/db","Treatment":"vehicle"},
"Additional sample data":{"RAW_FILE_NAME":"9"}
},
{
"Subject ID":"-",
"Sample ID":"21",
"Factors":{"Animal":"db/+","Treatment":"Cana"},
"Additional sample data":{"RAW_FILE_NAME":"21"}
},
{
"Subject ID":"-",
"Sample ID":"22",
"Factors":{"Animal":"db/+","Treatment":"Cana"},
"Additional sample data":{"RAW_FILE_NAME":"22"}
},
{
"Subject ID":"-",
"Sample ID":"23",
"Factors":{"Animal":"db/+","Treatment":"Cana"},
"Additional sample data":{"RAW_FILE_NAME":"23"}
},
{
"Subject ID":"-",
"Sample ID":"24",
"Factors":{"Animal":"db/+","Treatment":"Cana"},
"Additional sample data":{"RAW_FILE_NAME":"24"}
},
{
"Subject ID":"-",
"Sample ID":"5",
"Factors":{"Animal":"db/+","Treatment":"Cana"},
"Additional sample data":{"RAW_FILE_NAME":"5"}
},
{
"Subject ID":"-",
"Sample ID":"6",
"Factors":{"Animal":"db/+","Treatment":"Cana"},
"Additional sample data":{"RAW_FILE_NAME":"6"}
},
{
"Subject ID":"-",
"Sample ID":"7",
"Factors":{"Animal":"db/+","Treatment":"Cana"},
"Additional sample data":{"RAW_FILE_NAME":"7"}
},
{
"Subject ID":"-",
"Sample ID":"8",
"Factors":{"Animal":"db/+","Treatment":"Cana"},
"Additional sample data":{"RAW_FILE_NAME":"8"}
},
{
"Subject ID":"-",
"Sample ID":"1",
"Factors":{"Animal":"db/+","Treatment":"vehicle"},
"Additional sample data":{"RAW_FILE_NAME":"1"}
},
{
"Subject ID":"-",
"Sample ID":"17",
"Factors":{"Animal":"db/+","Treatment":"vehicle"},
"Additional sample data":{"RAW_FILE_NAME":"17"}
},
{
"Subject ID":"-",
"Sample ID":"18",
"Factors":{"Animal":"db/+","Treatment":"vehicle"},
"Additional sample data":{"RAW_FILE_NAME":"18"}
},
{
"Subject ID":"-",
"Sample ID":"19",
"Factors":{"Animal":"db/+","Treatment":"vehicle"},
"Additional sample data":{"RAW_FILE_NAME":"19"}
},
{
"Subject ID":"-",
"Sample ID":"2",
"Factors":{"Animal":"db/+","Treatment":"vehicle"},
"Additional sample data":{"RAW_FILE_NAME":"2"}
},
{
"Subject ID":"-",
"Sample ID":"20",
"Factors":{"Animal":"db/+","Treatment":"vehicle"},
"Additional sample data":{"RAW_FILE_NAME":"20"}
},
{
"Subject ID":"-",
"Sample ID":"3",
"Factors":{"Animal":"db/+","Treatment":"vehicle"},
"Additional sample data":{"RAW_FILE_NAME":"3"}
},
{
"Subject ID":"-",
"Sample ID":"4",
"Factors":{"Animal":"db/+","Treatment":"vehicle"},
"Additional sample data":{"RAW_FILE_NAME":"4"}
},
{
"Subject ID":"-",
"Sample ID":"49",
"Factors":{"Animal":"db/+","Treatment":"vehicle"},
"Additional sample data":{"RAW_FILE_NAME":"49"}
},
{
"Subject ID":"-",
"Sample ID":"50",
"Factors":{"Animal":"db/+","Treatment":"vehicle"},
"Additional sample data":{"RAW_FILE_NAME":"50"}
},
{
"Subject ID":"-",
"Sample ID":"51",
"Factors":{"Animal":"db/+","Treatment":"vehicle"},
"Additional sample data":{"RAW_FILE_NAME":"51"}
},
{
"Subject ID":"-",
"Sample ID":"52",
"Factors":{"Animal":"db/+","Treatment":"vehicle"},
"Additional sample data":{"RAW_FILE_NAME":"52"}
}
],
"COLLECTION":{"COLLECTION_SUMMARY":"All mice were housed in a pathogen-free facility and maintained on a 12-h light and 12-h dark cycle with free access to normal chow diet and water. CANA was mixed at 0.03% (w/w) with CRF-1 and administered to 8-week-old db/+ and db/db mice for 4 weeks. Food intake was calculated for 3 consecutive days each week using the mouse feeder MF-4S. At the end of the experiments, 6 h-fasted mice were anesthetized with isoflurane and blood samples were obtained from the inferior vena cava. Skeletal muscles including soleus and extensor digitorum longus were immediately frozen and stored at -80°C for metabolome and lipidome analysis.","SAMPLE_TYPE":"Skeletal muscles"},

"TREATMENT":{"TREATMENT_SUMMARY":"All mice were free access to normal chow diet and water. CANA was mixed at 0.03% (w/w) with CRF-1 and administered to 8-week-old db/+ and db/db mice for 4 weeks."},

"SAMPLEPREP":{"SAMPLEPREP_SUMMARY":"Metabolites were extracted from frozen and crushed skeletal muscle (~10 mg) or serum (20 uL) with 1 mL of solvent mixture (methanol:chloroform:water = 10:4:4, v/v/v) containing piperazine-1,4-bis (2-ethanesulfonic acid) (PIPES) (0.63 umol/L), 2-bromohypoxanthine (0.13 umol/L), free fatty acid (FA) 16:0 (13C16) (0.13 umol/L), diacylglycerol (DG) 15:0-18:1 (d7) (0.15 umol/L), and triacylglycerol (TG) 15:0-18:1 (d7) -15:0 (0.35 umol/L) as internal standards. The samples were vigorously mixed for 1 min and sonicated for 5 min. The extracts were then centrifuged at 16,000 × g for 5 min at 4°C, and the resultant supernatant was collected. Protein concentrations in the pellets were determined using a Pierce BCA Protein Assay Kit (Thermo Fisher Scientific, Massachusetts, USA). The collected supernatant (700 uL) was mixed with 235 uL of chloroform, and 155 uL of water, and then centrifuged at 16,000 × g for 5 min at 4°C. The aqueous (upper) layer (400 µL) was evaporated under vacuum, dried extracts were stored at −80°C until targeted polar metablome analysis. Prior to analysis, the dried aqueous layer was reconstituted in 50 uL of water. The organic (lower) layer (200 µL) was dried under a nitrogen stream, dried extracts were stored at −80°C until targeted lipidome analysis. Prior to analysis, the organic aqueous layer was reconstituted in 80 µL of methanol/chloroform (1/1, v/v)"},

"CHROMATOGRAPHY":{"CHROMATOGRAPHY_SUMMARY":"Cationic polar metabolites (i.e., amino acids, bases, nucleosides, etc) were analyzed via PFPP-LC/HRMS/MS.","INSTRUMENT_NAME":"Shimadzu Nexera X2","COLUMN_NAME":"Merck Discovery HS F5 (PFP) (150 x 2.1mm,3um)","COLUMN_TEMPERATURE":"40℃","FLOW_GRADIENT":"0%B, 0-5.0 min, 0-40%B, 5.0-15.0 min, 40-100%B, 15.0-15.1 min, 100%B, 15.1-18.0 min, 100-0%B, 18.0-18.1 min, 0%B, 18.1-25.0 min","FLOW_RATE":"0.25 mL/min","SOLVENT_A":"0.1% formic acid in H2O","SOLVENT_B":"ACN","CHROMATOGRAPHY_TYPE":"Reversed phase"},

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

"MS":{"INSTRUMENT_NAME":"Thermo Q Exactive Orbitrap","INSTRUMENT_TYPE":"Orbitrap","MS_TYPE":"ESI","MS_COMMENTS":"MS acquisition: FullMS (posi/nega) Data processing/software: Reifycs Cascade ver. 1.0","ION_MODE":"UNSPECIFIED"},

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

"Data":[{"Metabolite":"1-Methylhistidine","13":"0.4598","14":"0.3124","15":"0.6221","16":"0.3531","61":"1.1420","62":"1.4707","63":"1.7232","64":"2.7632","10":"0.2358","11":"0.2151","12":"0.2604","57":"1.3198","58":"2.0945","59":"2.8547","60":"2.0221","9":"0.2475","21":"0.2152","22":"0.2221","23":"0.1858","24":"0.3035","5":"0.2810","6":"0.3616","7":"0.3774","8":"0.2217","1":"0.6002","17":"0.1921","18":"0.3005","19":"0.2543","2":"0.5828","20":"0.2451","3":"1.1578","4":"0.5523","49":"3.1253","50":"4.2600","51":"2.0802","52":"2.1136"},{"Metabolite":"3-Cyano-L-alanine","13":"0.0119","14":"0.0111","15":"0.0085","16":"0.0114","61":"","62":"","63":"","64":"","10":"0.0232","11":"0.0232","12":"0.0182","57":"","58":"","59":"","60":"","9":"0.0244","21":"0.0093","22":"0.0107","23":"0.0099","24":"0.0140","5":"0.0192","6":"0.0241","7":"0.0173","8":"0.0198","1":"0.0258","17":"0.0076","18":"0.0086","19":"0.0036","2":"0.0184","20":"0.0081","3":"0.0234","4":"0.0206","49":"0.0052","50":"0.0202","51":"0.0097","52":"0.0121"},{"Metabolite":"4-Guanidinobutanoate","13":"0.0232","14":"0.0130","15":"0.0460","16":"0.0124","61":"","62":"","63":"0.0230","64":"","10":"0.1170","11":"0.0737","12":"0.0738","57":"","58":"0.1198","59":"0.2027","60":"","9":"0.0968","21":"0.0453","22":"0.0386","23":"0.0340","24":"0.0352","5":"0.1114","6":"0.1409","7":"0.1332","8":"0.0963","1":"0.0662","17":"0.0513","18":"0.0643","19":"0.0404","2":"0.0325","20":"0.0423","3":"0.1320","4":"0.0328","49":"","50":"","51":"0.0413","52":"0.0208"},{"Metabolite":"4-Hydroxy-L-proline","13":"2.7947","14":"2.3869","15":"1.3291","16":"2.6353","61":"0.1962","62":"0.2298","63":"0.2406","64":"0.2606","10":"0.9474","11":"0.6891","12":"0.7513","57":"0.4153","58":"0.6633","59":"1.0285","60":"0.4380","9":"0.7320","21":"0.6518","22":"0.7047","23":"0.9234","24":"0.7487","5":"0.7290","6":"1.4067","7":"1.0314","8":"0.8599","1":"1.0917","17":"1.2594","18":"1.2427","19":"1.3652","2":"1.1603","20":"1.3633","3":"0.7855","4":"0.9280","49":"2.3635","50":"2.8885","51":"0.2691","52":"0.6462"},{"Metabolite":"5^-Methylthioadenosine","13":"0.1397","14":"0.1178","15":"0.1470","16":"0.1358","61":"0.0371","62":"0.0372","63":"0.0429","64":"0.0667","10":"0.1173","11":"0.1220","12":"0.1255","57":"0.0237","58":"0.0376","59":"0.0378","60":"0.0337","9":"0.1297","21":"0.0979","22":"0.0890","23":"0.1143","24":"0.1275","5":"0.1334","6":"0.1390","7":"0.1470","8":"0.1058","1":"0.1404","17":"0.0783","18":"0.1393","19":"0.1376","2":"0.1666","20":"0.1306","3":"0.1285","4":"0.1185","49":"0.0652","50":"0.0458","51":"0.0430","52":"0.0392"},{"Metabolite":"Acetylglycine","13":"0.0933","14":"0.0493","15":"0.1789","16":"0.1176","61":"2.2796","62":"3.1419","63":"3.1405","64":"3.7250","10":"0.1457","11":"0.0892","12":"0.0718","57":"1.0771","58":"1.2843","59":"1.4211","60":"1.5782","9":"0.0728","21":"0.0527","22":"0.0834","23":"0.0523","24":"0.0691","5":"0.0590","6":"0.0474","7":"0.0770","8":"0.0407","1":"0.1123","17":"0.0503","18":"0.0387","19":"0.0257","2":"0.0664","20":"0.0295","3":"0.2318","4":"0.1272","49":"3.3144","50":"2.8443","51":"1.6076","52":"1.5789"},{"Metabolite":"Adenine","13":"0.1039","14":"0.0965","15":"0.1174","16":"0.1252","61":"","62":"","63":"","64":"","10":"0.0629","11":"0.0634","12":"0.0690","57":"","58":"","59":"","60":"","9":"0.0584","21":"0.0569","22":"0.0703","23":"0.0755","24":"0.0961","5":"0.0746","6":"0.1026","7":"0.0983","8":"0.0487","1":"0.0971","17":"0.0921","18":"0.0958","19":"0.1000","2":"0.0790","20":"0.1152","3":"0.0893","4":"0.0570","49":"0.1013","50":"","51":"","52":""},{"Metabolite":"Adenosine","13":"0.9537","14":"2.0738","15":"1.8614","16":"1.4367","61":"0.0677","62":"0.0921","63":"0.1288","64":"0.2912","10":"5.2064","11":"3.3085","12":"4.4708","57":"0.0939","58":"0.2089","59":"0.1895","60":"0.4928","9":"4.3047","21":"4.4588","22":"1.3075","23":"2.8322","24":"3.5041","5":"4.5615","6":"4.3422","7":"3.3068","8":"3.9858","1":"1.7992","17":"1.5165","18":"3.7030","19":"1.6998","2":"2.4836","20":"2.0615","3":"2.7642","4":"2.3136","49":"13.6396","50":"0.9901","51":"0.0900","52":"0.1096"},{"Metabolite":"Adenylosuccinic 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