#METABOLOMICS WORKBENCH tara600_20231221_193318 DATATRACK_ID:4548 STUDY_ID:ST003042 ANALYSIS_ID:AN004990 VERSION 1 CREATED_ON 01-16-2024 #PROJECT PR:PROJECT_TITLE The metabolomic resetting effect of DY131 in cisplatin-induced AKI PR:PROJECT_TYPE MS quantitative analysis PR:PROJECT_SUMMARY Acute kidney injury (AKI) remains a challenge in clinical practice, and PR:PROJECT_SUMMARY mitochondrial injury is a hallmark of AKI independent of the exact aetiology. PR:PROJECT_SUMMARY ERRγ is a member of orphan nuclear receptors which plays a regulatory role in PR:PROJECT_SUMMARY mitochondrial biosynthesis, energy metabolism, oxidative stress, cell apoptosis, PR:PROJECT_SUMMARY inflammation, and especially metabolic pathways. Here we investigate the role of PR:PROJECT_SUMMARY pharmacological agonist of ERRγ, DY131, in AKI mice induced by cisplatin, IR PR:PROJECT_SUMMARY and LPS. DY131 ameliorated renal function, tubular injury, cell apoptosis and PR:PROJECT_SUMMARY inflammation in AKI mice with multiple causes. Furthermore, we performed PR:PROJECT_SUMMARY LC-MS/MS analyses using renal tissues from cisplatin-induced AKI mice with or PR:PROJECT_SUMMARY without DY131 treatment. Strikingly, the data revealed that DY131 alleviated PR:PROJECT_SUMMARY cisplatin-induced mitochondrial dysfunction and energy metabolism disorder, as PR:PROJECT_SUMMARY well as multiple metabolic disorders. Taken together, the findings highlighted PR:PROJECT_SUMMARY the protective effect of DY131 on AKI probably via improving mitochondrial PR:PROJECT_SUMMARY function and energy metabolism. PR:INSTITUTE Children's Hospital of Nanjing Medical University PR:DEPARTMENT Department of Nephrology, State Key Laboratory of Reproductive Medicine PR:LABORATORY Nanjing Key Lab of Pediatrics, Jiangsu Key Laboratory of Pediatrics PR:LAST_NAME Lu PR:FIRST_NAME Lingling PR:ADDRESS Guangzhou Road 72, Nanjing, Jiangsu, 210000, China PR:EMAIL lulingling89tara@163.com PR:PHONE 0086-25-8311-7435 PR:DOI http://dx.doi.org/10.21228/M81F0P #STUDY ST:STUDY_TITLE The metabolomic resetting effect of DY131 in cisplatin-induced AKI ST:STUDY_TYPE MS ST:STUDY_SUMMARY LC-MS/MS analyses were performed using renal tissues from cisplatin-induced AKI ST:STUDY_SUMMARY mice with or without DY131 treatment. The data revealed that DY131 alleviated ST:STUDY_SUMMARY cisplatin-induced mitochondrial dysfunction and energy metabolism disorder, as ST:STUDY_SUMMARY well as multiple metabolic disorders. ST:INSTITUTE Children's Hospital of Nanjing Medical University ST:DEPARTMENT Department of Nephrology, State Key Laboratory of Reproductive Medicine ST:LABORATORY Nanjing Key Lab of Pediatrics, Jiangsu Key Laboratory of Pediatrics ST:LAST_NAME Lu ST:FIRST_NAME Lingling ST:ADDRESS Guangzhou Road 72, Nanjing, Jiangsu, 210000, China ST:EMAIL lulingling89tara@163.com ST:PHONE 0086-25-8311-7435 ST:SUBMIT_DATE 2023-12-21 #SUBJECT SU:SUBJECT_TYPE Mammal SU:SUBJECT_SPECIES Mus musculus SU:TAXONOMY_ID 10090 SU:GENOTYPE_STRAIN C57BL/6J SU:AGE_OR_AGE_RANGE 7-8 weeks SU:WEIGHT_OR_WEIGHT_RANGE 20-25g SU:GENDER Male SU:ANIMAL_ANIMAL_SUPPLIER Nanjing Medical University SU:ANIMAL_HOUSING Nanjing Medical University SU:ANIMAL_FEED Free to food SU:ANIMAL_WATER Free to clean water #SUBJECT_SAMPLE_FACTORS: SUBJECT(optional)[tab]SAMPLE[tab]FACTORS(NAME:VALUE pairs separated by |)[tab]Additional sample data SUBJECT_SAMPLE_FACTORS - C_1 Treatment:control RAW_FILE_NAME=pos_C_1.mzXML; RAW_FILE_NAME=neg_C_1.mzXML SUBJECT_SAMPLE_FACTORS - C_2 Treatment:control RAW_FILE_NAME=pos_C_2.mzXML; RAW_FILE_NAME=neg_C_2.mzXML SUBJECT_SAMPLE_FACTORS - C_3 Treatment:control RAW_FILE_NAME=pos_C_3.mzXML; RAW_FILE_NAME=neg_C_3.mzXML SUBJECT_SAMPLE_FACTORS - C_4 Treatment:control RAW_FILE_NAME=pos_C_4.mzXML; RAW_FILE_NAME=neg_C_4.mzXML SUBJECT_SAMPLE_FACTORS - C_5 Treatment:control RAW_FILE_NAME=pos_C_5.mzXML; RAW_FILE_NAME=neg_C_5.mzXML SUBJECT_SAMPLE_FACTORS - C_6 Treatment:control RAW_FILE_NAME=pos_C_6.mzXML; RAW_FILE_NAME=neg_C_6.mzXML SUBJECT_SAMPLE_FACTORS - C_7 Treatment:control RAW_FILE_NAME=pos_C_7.mzXML; RAW_FILE_NAME=neg_C_7.mzXML SUBJECT_SAMPLE_FACTORS - DY_P_1 Treatment:DY131+cisplatin RAW_FILE_NAME=pos_DP_1.mzXML; RAW_FILE_NAME=neg_DP_1.mzXML SUBJECT_SAMPLE_FACTORS - DY_P_2 Treatment:DY131+cisplatin RAW_FILE_NAME=pos_DP_2.mzXML; RAW_FILE_NAME=neg_DP_2.mzXML SUBJECT_SAMPLE_FACTORS - DY_P_3 Treatment:DY131+cisplatin RAW_FILE_NAME=pos_DP_3.mzXML; RAW_FILE_NAME=neg_DP_3.mzXML SUBJECT_SAMPLE_FACTORS - DY_P_4 Treatment:DY131+cisplatin RAW_FILE_NAME=pos_DP_4.mzXML; RAW_FILE_NAME=neg_DP_4.mzXML SUBJECT_SAMPLE_FACTORS - DY_P_5 Treatment:DY131+cisplatin RAW_FILE_NAME=pos_DP_5.mzXML; RAW_FILE_NAME=neg_DP_5.mzXML SUBJECT_SAMPLE_FACTORS - DY_P_6 Treatment:DY131+cisplatin RAW_FILE_NAME=pos_DP_6.mzXML; RAW_FILE_NAME=neg_DP_6.mzXML SUBJECT_SAMPLE_FACTORS - DY_P_7 Treatment:DY131+cisplatin RAW_FILE_NAME=pos_DP_7.mzXML; RAW_FILE_NAME=neg_DP_7.mzXML SUBJECT_SAMPLE_FACTORS - P_1 Treatment:vehicle+cisplatin RAW_FILE_NAME=pos_P_1.mzXML; RAW_FILE_NAME=neg_P_1.mzXML SUBJECT_SAMPLE_FACTORS - P_2 Treatment:vehicle+cisplatin RAW_FILE_NAME=pos_P_2.mzXML; RAW_FILE_NAME=neg_P_2.mzXML SUBJECT_SAMPLE_FACTORS - P_3 Treatment:vehicle+cisplatin RAW_FILE_NAME=pos_P_3.mzXML; RAW_FILE_NAME=neg_P_3.mzXML SUBJECT_SAMPLE_FACTORS - P_4 Treatment:vehicle+cisplatin RAW_FILE_NAME=pos_P_4.mzXML; RAW_FILE_NAME=neg_P_4.mzXML SUBJECT_SAMPLE_FACTORS - P_5 Treatment:vehicle+cisplatin RAW_FILE_NAME=pos_P_5.mzXML; RAW_FILE_NAME=neg_P_5.mzXML SUBJECT_SAMPLE_FACTORS - P_6 Treatment:vehicle+cisplatin RAW_FILE_NAME=pos_P_6.mzXML; RAW_FILE_NAME=neg_P_6.mzXML SUBJECT_SAMPLE_FACTORS - P_7 Treatment:vehicle+cisplatin RAW_FILE_NAME=pos_P_7.mzXML; RAW_FILE_NAME=neg_P_7.mzXML #COLLECTION CO:COLLECTION_SUMMARY The mice were euthanized 72 hours after cisplatin injection, and renal tissues CO:COLLECTION_SUMMARY were collected and frozen at -80℃. CO:SAMPLE_TYPE Kidney CO:STORAGE_CONDITIONS -80℃ #TREATMENT TR:TREATMENT_SUMMARY Wild type C57BL/6J mice were maintained under standard environmental conditions. TR:TREATMENT_SUMMARY A single injection of cisplatin (25 mg/kg, i.p.) was used to induce AKI. DY131 TR:TREATMENT_SUMMARY was dissolved in DMSO and further diluted with 5% Tween-80. To assess the effect TR:TREATMENT_SUMMARY of DY131, the mice were injected with DY131 (5 mg/kg, i.p.) or vehicle one hour TR:TREATMENT_SUMMARY before cisplatin injection, repeated every 24 hours. The mice were euthanized 72 TR:TREATMENT_SUMMARY hours after cisplatin injection. TR:TREATMENT_COMPOUND cisplatin or saline, DY131 or vehicle TR:TREATMENT_ROUTE intraperitoneal injection ​ TR:TREATMENT_DOSE cisplatin at 25 mg/kg, once. DY131 at 5 mg/kg, once a day for 3 days TR:TREATMENT_DOSEVOLUME Animal Vet Treatments: Animal Anesthesia: Animal Acclimation Duration: Animal TR:TREATMENT_DOSEVOLUME Fasting: Animal Endpoint Euthanasia: Animal Endpoint Tissue Coll. List: TR:TREATMENT_DOSEVOLUME Animal Endpoint Tissue Proc. Method: Animal Endpoint Clinical Signs: TR:TREATMENT_VEHICLE cisplatin dissolved in saline, DY131 dissolved in DMSO and further diluted with TR:TREATMENT_VEHICLE 5% Tween-80 #SAMPLEPREP SP:SAMPLEPREP_SUMMARY 25 mg of sample was weighted to an EP tube, and 500 μL extract solution SP:SAMPLEPREP_SUMMARY (acetonitrile: methanol: water = 2: 2: 1, with isotopically-labelled internal SP:SAMPLEPREP_SUMMARY standard mixture) was added. After 30 s vortex, the samples were homogenized at SP:SAMPLEPREP_SUMMARY 35 Hz for 4 min and sonicated for 5 min in ice-water bath. The homogenization SP:SAMPLEPREP_SUMMARY and sonication cycle was repeated for 3 times. Then the samples were incubated SP:SAMPLEPREP_SUMMARY for 1 h at -40 ℃ and centrifuged at 12000 rpm for 15 min at 4 ℃. The SP:SAMPLEPREP_SUMMARY resulting supernatant was transferred to a fresh glass vial for analysis. #CHROMATOGRAPHY CH:METHODS_FILENAME DY131_Chromatography.txt CH:INSTRUMENT_NAME Thermo Fisher Scientific CH:COLUMN_NAME Waters ACQUITY UPLC BEH Amide (100 x 2.1mm,1.7um) CH:COLUMN_TEMPERATURE 30 ℃ CH:FLOW_GRADIENT 0-0.5min, 95% B; 0.5-7min, 95%-65%B; 7-8min, 65%B-40%B; 8-9min, 40%B; 9-9.1min, CH:FLOW_GRADIENT 40%-95%B; 9.1-12min, 95%B CH:FLOW_RATE 0.5ml/min CH:SOLVENT_A water（pH = 9.75) CH:SOLVENT_B acetonitrile CH:CHROMATOGRAPHY_TYPE HILIC #ANALYSIS AN:ANALYSIS_TYPE MS AN:ANALYSIS_PROTOCOL_FILE DY131_MS.txt #MS MS:INSTRUMENT_NAME Thermo Q Exactive HF-X Orbitrap MS:INSTRUMENT_TYPE Orbitrap MS:MS_TYPE ESI MS:MS_COMMENTS sheath gas flow rate as 30 Arb, Aux gas flow rate as 25 Arb, capillary MS:MS_COMMENTS temperature 350 ℃, full MS resolution as 60000, MS/MS resolution as 7500, MS:MS_COMMENTS collision energy as 10/30/60 in NCE mode, spray Voltage as 3.6 kV (positive). MS:MS_COMMENTS Metabolites were quantified by relative quantification and expressed by peak MS:MS_COMMENTS area. MS:ION_MODE POSITIVE MS:MS_RESULTS_FILE ST003042_AN004990_Results.txt UNITS:Peak area Has m/z:Yes Has RT:Yes RT units:Minutes #END