#METABOLOMICS WORKBENCH ZJ309_20220719_052137 DATATRACK_ID:3356 STUDY_ID:ST002384 ANALYSIS_ID:AN003883 PROJECT_ID:PR001534 VERSION 1 CREATED_ON October 16, 2022, 5:40 am #PROJECT PR:PROJECT_TITLE Lipidomics Investigation of Curcumin and Epigallocatechin Gallate on PR:PROJECT_TITLE IgE-Mediated Degranulation of RBL-2H3 Cells PR:PROJECT_SUMMARY In this study, an IgE-mediated RBL-2H3 cell degranulation model was firstly PR:PROJECT_SUMMARY developed. Then, ultra-high-performance liquid chromatography coupled to mass PR:PROJECT_SUMMARY spectrometry (UPLC-MS) was used to perform the non-targeted lipidomics study, in PR:PROJECT_SUMMARY order to investigate the effects and mechanism of curcumin/EGCG on IgE-mediated PR:PROJECT_SUMMARY RBL-2H3 cells degranulation. PR:INSTITUTE College of Marine Food and Biological Engineering, Jimei University PR:LAST_NAME Yang PR:FIRST_NAME Zhiqiang PR:ADDRESS No. 185 Yinjiang Road, Jimei District, Xiamen City, Fujian Province, China PR:EMAIL 201911832016@jmu.edu.cn PR:PHONE 0592-6181487 #STUDY ST:STUDY_TITLE Lipidomics study of curcumin and epigallocatechin gallate on IgE-mediated ST:STUDY_TITLE degranulation of RBL-2H3 cells ST:STUDY_SUMMARY This study provides novel insights into the potential anti-allergic mechanism of ST:STUDY_SUMMARY food-derived curcumin and EGCG, and then facilitates the discovery of drug ST:STUDY_SUMMARY target and the development of diagnostic methods for allergic diseases. ST:INSTITUTE College of Marine Food and Biological Engineering, Jimei University ST:LAST_NAME Yang ST:FIRST_NAME Zhiqiang ST:ADDRESS No. 185 Yinjiang Road, Jimei District, Xiamen City, Fujian Province, China ST:EMAIL 201911832016@jmu.edu.cn ST:PHONE 0592-6181487 #SUBJECT SU:SUBJECT_TYPE Cultured cells SU:SUBJECT_SPECIES Rattus norvegicus SU:TAXONOMY_ID 10116 #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 - PBS-1A Treatment:Control Model=Positive; RAW_FILE_NAME=PBS-1A.RAW SUBJECT_SAMPLE_FACTORS - PBS-1B Treatment:Control Model=Positive; RAW_FILE_NAME=PBS-1B.RAW SUBJECT_SAMPLE_FACTORS - PBS-1C Treatment:Control Model=Positive; RAW_FILE_NAME=PBS-1C.RAW SUBJECT_SAMPLE_FACTORS - PBS-1D Treatment:Control Model=Positive; RAW_FILE_NAME=PBS-1D.RAW SUBJECT_SAMPLE_FACTORS - 0-1A Treatment:Allergy group Model=Positive; RAW_FILE_NAME=0-1A.RAW SUBJECT_SAMPLE_FACTORS - 0-1B Treatment:Allergy group Model=Positive; RAW_FILE_NAME=0-1B.RAW SUBJECT_SAMPLE_FACTORS - 0-1C Treatment:Allergy group Model=Positive; RAW_FILE_NAME=0-1C.RAW SUBJECT_SAMPLE_FACTORS - 0-1D Treatment:Allergy group Model=Positive; RAW_FILE_NAME=0-1D.RAW SUBJECT_SAMPLE_FACTORS - C10-1A Treatment:After allergy, 10uM curcumin interventionfor 1h Model=Positive; RAW_FILE_NAME=C10-1A.RAW SUBJECT_SAMPLE_FACTORS - C10-1B Treatment:After allergy, 10uM curcumin interventionfor 1h Model=Positive; RAW_FILE_NAME=C10-1B.RAW SUBJECT_SAMPLE_FACTORS - C10-1C Treatment:After allergy, 10uM curcumin interventionfor 1h Model=Positive; RAW_FILE_NAME=C10-1C.RAW SUBJECT_SAMPLE_FACTORS - C10-1D Treatment:After allergy, 10uM curcumin interventionfor 1h Model=Positive; RAW_FILE_NAME=C10-1D.RAW SUBJECT_SAMPLE_FACTORS - C30-1A Treatment:After allergy, 30uM curcumin interventionfor 1h Model=Positive; RAW_FILE_NAME=C30-1A.RAW SUBJECT_SAMPLE_FACTORS - C30-1B Treatment:After allergy, 30uM curcumin interventionfor 1h Model=Positive; RAW_FILE_NAME=C30-1B.RAW SUBJECT_SAMPLE_FACTORS - C30-1C Treatment:After allergy, 30uM curcumin interventionfor 1h Model=Positive; RAW_FILE_NAME=C30-1C.RAW SUBJECT_SAMPLE_FACTORS - C30-1D Treatment:After allergy, 30uM curcumin interventionfor 1h Model=Positive; RAW_FILE_NAME=C30-1D.RAW SUBJECT_SAMPLE_FACTORS - E200-1A Treatment:After allergy, 200uM EGCG interventionfor 1h Model=Positive; RAW_FILE_NAME=E200-1A.RAW SUBJECT_SAMPLE_FACTORS - E200-1B Treatment:After allergy, 200uM EGCG interventionfor 1h Model=Positive; RAW_FILE_NAME=E200-1B.RAW SUBJECT_SAMPLE_FACTORS - E200-1C Treatment:After allergy, 200uM EGCG interventionfor 1h Model=Positive; RAW_FILE_NAME=E200-1C.RAW SUBJECT_SAMPLE_FACTORS - E200-1D Treatment:After allergy, 200uM EGCG interventionfor 1h Model=Positive; RAW_FILE_NAME=E200-1D.RAW SUBJECT_SAMPLE_FACTORS - E500-1A Treatment:After allergy, 500uM EGCG interventionfor 1h Model=Positive; RAW_FILE_NAME=E500-1A.RAW SUBJECT_SAMPLE_FACTORS - E500-1B Treatment:After allergy, 500uM EGCG interventionfor 1h Model=Positive; RAW_FILE_NAME=E500-1B.RAW SUBJECT_SAMPLE_FACTORS - E500-1C Treatment:After allergy, 500uM EGCG interventionfor 1h Model=Positive; RAW_FILE_NAME=E500-1C.RAW SUBJECT_SAMPLE_FACTORS - E500-1D Treatment:After allergy, 500uM EGCG interventionfor 1h Model=Positive; RAW_FILE_NAME=E500-1D.RAW SUBJECT_SAMPLE_FACTORS - PBS-3A Treatment:Control Model=Positive; RAW_FILE_NAME=PBS-3A.RAW SUBJECT_SAMPLE_FACTORS - PBS-3B Treatment:Control Model=Positive; RAW_FILE_NAME=PBS-3B.RAW SUBJECT_SAMPLE_FACTORS - PBS-3C Treatment:Control Model=Positive; RAW_FILE_NAME=PBS-3C.RAW SUBJECT_SAMPLE_FACTORS - PBS-3D Treatment:Control Model=Positive; RAW_FILE_NAME=PBS-3D.RAW SUBJECT_SAMPLE_FACTORS - 0-3A Treatment:Allergy group Model=Positive; RAW_FILE_NAME=0-3A.RAW SUBJECT_SAMPLE_FACTORS - 0-3B Treatment:Allergy group Model=Positive; RAW_FILE_NAME=0-3B.RAW SUBJECT_SAMPLE_FACTORS - 0-3C Treatment:Allergy group Model=Positive; RAW_FILE_NAME=0-3C.RAW SUBJECT_SAMPLE_FACTORS - 0-3D Treatment:Allergy group Model=Positive; RAW_FILE_NAME=0-3D.RAW SUBJECT_SAMPLE_FACTORS - C10-3A Treatment:After allergy, 10uM curcumin interventionfor 3h Model=Positive; RAW_FILE_NAME=C10-3A.RAW SUBJECT_SAMPLE_FACTORS - C10-3B Treatment:After allergy, 10uM curcumin interventionfor 3h Model=Positive; RAW_FILE_NAME=C10-3B.RAW SUBJECT_SAMPLE_FACTORS - C10-3C Treatment:After allergy, 10uM curcumin interventionfor 3h Model=Positive; RAW_FILE_NAME=C10-3C.RAW SUBJECT_SAMPLE_FACTORS - C10-3D Treatment:After allergy, 10uM curcumin interventionfor 3h Model=Positive; RAW_FILE_NAME=C10-3D.RAW SUBJECT_SAMPLE_FACTORS - C30-3A Treatment:After allergy, 30uM curcumin interventionfor 3h Model=Positive; RAW_FILE_NAME=C30-3A.RAW SUBJECT_SAMPLE_FACTORS - C30-3B Treatment:After allergy, 30uM curcumin interventionfor 3h Model=Positive; RAW_FILE_NAME=C30-3B.RAW SUBJECT_SAMPLE_FACTORS - C30-3C Treatment:After allergy, 30uM curcumin interventionfor 3h Model=Positive; RAW_FILE_NAME=C30-3C.RAW SUBJECT_SAMPLE_FACTORS - C30-3D Treatment:After allergy, 30uM curcumin interventionfor 3h Model=Positive; RAW_FILE_NAME=C30-3D.RAW SUBJECT_SAMPLE_FACTORS - E200-3A Treatment:After allergy, 200uM EGCG interventionfor 3h Model=Positive; RAW_FILE_NAME=E200-3A.RAW SUBJECT_SAMPLE_FACTORS - E200-3B Treatment:After allergy, 200uM EGCG interventionfor 3h Model=Positive; RAW_FILE_NAME=E200-3B.RAW SUBJECT_SAMPLE_FACTORS - E200-3C Treatment:After allergy, 200uM EGCG interventionfor 3h Model=Positive; RAW_FILE_NAME=E200-3C.RAW SUBJECT_SAMPLE_FACTORS - E200-3D Treatment:After allergy, 200uM EGCG interventionfor 3h Model=Positive; RAW_FILE_NAME=E200-3D.RAW SUBJECT_SAMPLE_FACTORS - E500-3A Treatment:After allergy, 500uM EGCG interventionfor 3h Model=Positive; RAW_FILE_NAME=E500-3A.RAW SUBJECT_SAMPLE_FACTORS - E500-3B Treatment:After allergy, 500uM EGCG interventionfor 3h Model=Positive; RAW_FILE_NAME=E500-3B.RAW SUBJECT_SAMPLE_FACTORS - E500-3C Treatment:After allergy, 500uM EGCG interventionfor 3h Model=Positive; RAW_FILE_NAME=E500-3C.RAW SUBJECT_SAMPLE_FACTORS - E500-3D Treatment:After allergy, 500uM EGCG interventionfor 3h Model=Positive; RAW_FILE_NAME=E500-3D.RAW SUBJECT_SAMPLE_FACTORS - PBS-1A-Negatve Treatment:Control Model=Negative; RAW_FILE_NAME=PBS-1A-Negatve.RAW SUBJECT_SAMPLE_FACTORS - PBS-1B-N Treatment:Control Model=Negative; RAW_FILE_NAME=PBS-1B-N.RAW SUBJECT_SAMPLE_FACTORS - PBS-1C-N Treatment:Control Model=Negative; RAW_FILE_NAME=PBS-1C-N.RAW SUBJECT_SAMPLE_FACTORS - PBS-1D-N Treatment:Control Model=Negative; RAW_FILE_NAME=PBS-1D-N.RAW SUBJECT_SAMPLE_FACTORS - 0-1A-N Treatment:Allergy group Model=Negative; RAW_FILE_NAME=0-1A-N.RAW SUBJECT_SAMPLE_FACTORS - 0-1B-N Treatment:Allergy group Model=Negative; RAW_FILE_NAME=0-1B-N.RAW SUBJECT_SAMPLE_FACTORS - 0-1C-N Treatment:Allergy group Model=Negative; RAW_FILE_NAME=0-1C-N.RAW SUBJECT_SAMPLE_FACTORS - 0-1D-N Treatment:Allergy group Model=Negative; RAW_FILE_NAME=0-1D-N.RAW SUBJECT_SAMPLE_FACTORS - C10-1A-N Treatment:After allergy, 10uM curcumin interventionfor 1h Model=Negative; RAW_FILE_NAME=C10-1A-N.RAW SUBJECT_SAMPLE_FACTORS - C10-1B-N Treatment:After allergy, 10uM curcumin interventionfor 1h Model=Negative; RAW_FILE_NAME=C10-1B-N.RAW SUBJECT_SAMPLE_FACTORS - C10-1C-N Treatment:After allergy, 10uM curcumin interventionfor 1h Model=Negative; RAW_FILE_NAME=C10-1C-N.RAW SUBJECT_SAMPLE_FACTORS - C10-1D-N Treatment:After allergy, 10uM curcumin interventionfor 1h Model=Negative; RAW_FILE_NAME=C10-1D-N.RAW SUBJECT_SAMPLE_FACTORS - C30-1A-N Treatment:After allergy, 30uM curcumin interventionfor 1h Model=Negative; RAW_FILE_NAME=C30-1A-N.RAW SUBJECT_SAMPLE_FACTORS - C30-1B-N Treatment:After allergy, 30uM curcumin interventionfor 1h Model=Negative; RAW_FILE_NAME=C30-1B-N.RAW SUBJECT_SAMPLE_FACTORS - C30-1C-N Treatment:After allergy, 30uM curcumin interventionfor 1h Model=Negative; RAW_FILE_NAME=C30-1C-N.RAW SUBJECT_SAMPLE_FACTORS - C30-1D-N Treatment:After allergy, 30uM curcumin interventionfor 1h Model=Negative; RAW_FILE_NAME=C30-1D-N.RAW SUBJECT_SAMPLE_FACTORS - E200-1A-N Treatment:After allergy, 200uM EGCG interventionfor 1h Model=Negative; RAW_FILE_NAME=E200-1A-N.RAW SUBJECT_SAMPLE_FACTORS - E200-1B-N Treatment:After allergy, 200uM EGCG interventionfor 1h Model=Negative; RAW_FILE_NAME=E200-1B-N.RAW SUBJECT_SAMPLE_FACTORS - E200-1C-N Treatment:After allergy, 200uM EGCG interventionfor 1h Model=Negative; RAW_FILE_NAME=E200-1C-N.RAW SUBJECT_SAMPLE_FACTORS - E200-1D-N Treatment:After allergy, 200uM EGCG interventionfor 1h Model=Negative; RAW_FILE_NAME=E200-1D-N.RAW SUBJECT_SAMPLE_FACTORS - E500-1A-N Treatment:After allergy, 500uM EGCG interventionfor 1h Model=Negative; RAW_FILE_NAME=E500-1A-N.RAW SUBJECT_SAMPLE_FACTORS - E500-1B-N Treatment:After allergy, 500uM EGCG interventionfor 1h Model=Negative; RAW_FILE_NAME=E500-1B-N.RAW SUBJECT_SAMPLE_FACTORS - E500-1C-N Treatment:After allergy, 500uM EGCG interventionfor 1h Model=Negative; RAW_FILE_NAME=E500-1C-N.RAW SUBJECT_SAMPLE_FACTORS - E500-1D-N Treatment:After allergy, 500uM EGCG interventionfor 1h Model=Negative; RAW_FILE_NAME=E500-1D-N.RAW SUBJECT_SAMPLE_FACTORS - PBS-3A-N Treatment:Control Model=Negative; RAW_FILE_NAME=PBS-3A-N.RAW SUBJECT_SAMPLE_FACTORS - PBS-3B-N Treatment:Control Model=Negative; RAW_FILE_NAME=PBS-3B-N.RAW SUBJECT_SAMPLE_FACTORS - PBS-3C-N Treatment:Control Model=Negative; RAW_FILE_NAME=PBS-3C-N.RAW SUBJECT_SAMPLE_FACTORS - PBS-3D-N Treatment:Control Model=Negative; RAW_FILE_NAME=PBS-3D-N.RAW SUBJECT_SAMPLE_FACTORS - 0-3A-N Treatment:Allergy group Model=Negative; RAW_FILE_NAME=0-3A-N.RAW SUBJECT_SAMPLE_FACTORS - 0-3B-N Treatment:Allergy group Model=Negative; RAW_FILE_NAME=0-3B-N.RAW SUBJECT_SAMPLE_FACTORS - 0-3C-N Treatment:Allergy group Model=Negative; RAW_FILE_NAME=0-3C-N.RAW SUBJECT_SAMPLE_FACTORS - 0-3D-N Treatment:Allergy group Model=Negative; RAW_FILE_NAME=0-3D-N.RAW SUBJECT_SAMPLE_FACTORS - C10-3A-N Treatment:After allergy, 10uM curcumin interventionfor 3h Model=Negative; RAW_FILE_NAME=C10-3A-N.RAW SUBJECT_SAMPLE_FACTORS - C10-3B-N Treatment:After allergy, 10uM curcumin interventionfor 3h Model=Negative; RAW_FILE_NAME=C10-3B-N.RAW SUBJECT_SAMPLE_FACTORS - C10-3C-N Treatment:After allergy, 10uM curcumin interventionfor 3h Model=Negative; RAW_FILE_NAME=C10-3C-N.RAW SUBJECT_SAMPLE_FACTORS - C10-3D-N Treatment:After allergy, 10uM curcumin interventionfor 3h Model=Negative; RAW_FILE_NAME=C10-3D-N.RAW SUBJECT_SAMPLE_FACTORS - C30-3A-N Treatment:After allergy, 30uM curcumin interventionfor 3h Model=Negative; RAW_FILE_NAME=C30-3A-N.RAW SUBJECT_SAMPLE_FACTORS - C30-3B-N Treatment:After allergy, 30uM curcumin interventionfor 3h Model=Negative; RAW_FILE_NAME=C30-3B-N.RAW SUBJECT_SAMPLE_FACTORS - C30-3C-N Treatment:After allergy, 30uM curcumin interventionfor 3h Model=Negative; RAW_FILE_NAME=C30-3C-N.RAW SUBJECT_SAMPLE_FACTORS - C30-3D-N Treatment:After allergy, 30uM curcumin interventionfor 3h Model=Negative; RAW_FILE_NAME=C30-3D-N.RAW SUBJECT_SAMPLE_FACTORS - E200-3A-N Treatment:After allergy, 200uM EGCG interventionfor 3h Model=Negative; RAW_FILE_NAME=E200-3A-N.RAW SUBJECT_SAMPLE_FACTORS - E200-3B-N Treatment:After allergy, 200uM EGCG interventionfor 3h Model=Negative; RAW_FILE_NAME=E200-3B-N.RAW SUBJECT_SAMPLE_FACTORS - E200-3C-N Treatment:After allergy, 200uM EGCG interventionfor 3h Model=Negative; RAW_FILE_NAME=E200-3C-N.RAW SUBJECT_SAMPLE_FACTORS - E200-3D-N Treatment:After allergy, 200uM EGCG interventionfor 3h Model=Negative; RAW_FILE_NAME=E200-3D-N.RAW SUBJECT_SAMPLE_FACTORS - E500-3A-N Treatment:After allergy, 500uM EGCG interventionfor 3h Model=Negative; RAW_FILE_NAME=E500-3A-N.RAW SUBJECT_SAMPLE_FACTORS - E500-3B-N Treatment:After allergy, 500uM EGCG interventionfor 3h Model=Negative; RAW_FILE_NAME=E500-3B-N.RAW SUBJECT_SAMPLE_FACTORS - E500-3C-N Treatment:After allergy, 500uM EGCG interventionfor 3h Model=Negative; RAW_FILE_NAME=E500-3C-N.RAW SUBJECT_SAMPLE_FACTORS - E500-3D-N Treatment:After allergy, 500uM EGCG interventionfor 3h Model=Negative; RAW_FILE_NAME=E500-3D-N.RAW #COLLECTION CO:COLLECTION_SUMMARY RBL-2H3 cells were incubated with 200 ng/mL of IgE for 18 h, followed by CO:COLLECTION_SUMMARY co-stimulation with 500 ng/mL DNP-BSA for the Alg group, polyphenolic substances CO:COLLECTION_SUMMARY (curcumin or EGCG) and 500 ng/mL DNP-BSA for the intervention group, and the CO:COLLECTION_SUMMARY same volume of PBS solution for the PBS group. The cell samples were collected CO:COLLECTION_SUMMARY at two different time points, 1 h and 3 h after the effect, respectively. CO:SAMPLE_TYPE Cultured cells #TREATMENT TR:TREATMENT_SUMMARY RBL-2H3 cells were incubated with 200 ng/mL of IgE for 18 h, followed by TR:TREATMENT_SUMMARY co-stimulation with 500 ng/mL DNP-BSA for the Alg group, polyphenolic substances TR:TREATMENT_SUMMARY (curcumin or EGCG) and 500 ng/mL DNP-BSA for the intervention group, and the TR:TREATMENT_SUMMARY same volume of PBS solution for the PBS group. #SAMPLEPREP SP:SAMPLEPREP_SUMMARY The MeOH/MTBE/H2O system was used for the extraction of lipid metabolites from SP:SAMPLEPREP_SUMMARY cell samples. Cell culture dishes stored at -80 °C were first removed on ice SP:SAMPLEPREP_SUMMARY and 1 mL of methanolic solution containing internal standards was added, SP:SAMPLEPREP_SUMMARY consisting of 1 μg/mL FFA (C16:0)-d3 and FFA (C18:0)-d3, 0.87 μg/mL LPC (19:0) SP:SAMPLEPREP_SUMMARY and SM (d18:1/12:0), 1.12 μg/mL PC (38: 0), 0.92 μg/mL PE (30:0), 0.67 μg/mL SP:SAMPLEPREP_SUMMARY TG (45:0), and 0.47 μg/mL Cer (d18:1/17:0). The cells were transferred to 5 mL SP:SAMPLEPREP_SUMMARY Eppendorf centrifuge tubes by scraping with a cell scraper, adding 2.5 mL of SP:SAMPLEPREP_SUMMARY MTBE and vortexing for 30 s. The cells were further shaken for 30 min at 10 °C SP:SAMPLEPREP_SUMMARY and 1000 rpm using a thermostatic mixer. 750 μL of ultrapure water was added, SP:SAMPLEPREP_SUMMARY vortexed for 60 s and allowed to stand for 5 min, and then centrifuged at 12,000 SP:SAMPLEPREP_SUMMARY g for 10 min at 6 °C. After separation of the two phases, 400 μL of the SP:SAMPLEPREP_SUMMARY hydrophobic phase was collected and dried in a cryogenic vacuum centrifuge SP:SAMPLEPREP_SUMMARY concentrator, and the resulting lyophilized sample was stored at -80 °C. #CHROMATOGRAPHY CH:CHROMATOGRAPHY_TYPE Reversed phase CH:INSTRUMENT_NAME Thermo Dionex Ultimate 3000 CH:COLUMN_NAME Waters Acquity BEH C8 (100 x 2.1mm, 1.7um) #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:INSTRUMENT_NAME Thermo Q Exactive HF-X Orbitrap MS:INSTRUMENT_TYPE Orbitrap MS:MS_TYPE ESI MS:ION_MODE POSITIVE MS:MS_COMMENTS Xcalibular MS:MS_RESULTS_FILE ST002384_AN003883_Results.txt UNITS:Peak area Has m/z:Yes Has RT:Yes RT units:Minutes #END