#METABOLOMICS WORKBENCH rioro_20230629_060334 DATATRACK_ID:4126 STUDY_ID:ST002765 ANALYSIS_ID:AN004501 PROJECT_ID:PR001723 VERSION 1 CREATED_ON June 30, 2023, 9:42 am #PROJECT PR:PROJECT_TITLE Lipid metabolism affects fetal development PR:PROJECT_SUMMARY Gestational asthma interferes with lipidomic metabolism of amniotic fluid and PR:PROJECT_SUMMARY fetal alveolar lavage fluid, thus inhibiting fetal development PR:INSTITUTE Nanjing University of Chinese Medicine PR:LAST_NAME Fang PR:FIRST_NAME Huafeng PR:ADDRESS No.138 xianlin road, nanjing city, Nanjing, China, 210046, China PR:EMAIL Riorofhf@outlook.com PR:PHONE +86 18852416998 #STUDY ST:STUDY_TITLE Disorder of Lipids Induced by Gestational Asthma and its Effect on the ST:STUDY_TITLE Development of Fetal Lung Function and Fetal Health ST:STUDY_SUMMARY Maternal asthma during pregnancy is highly correlated with fetal growth and ST:STUDY_SUMMARY development, and can cause damage to both the mother and fetus, but the ST:STUDY_SUMMARY underlying mechanisms are not yet clear. Amniotic fluid, as the environment for ST:STUDY_SUMMARY fetal growth and development, may be affected by lipid metabolism disorders, ST:STUDY_SUMMARY which can impact fetal lung function development. A rat model of asthma during ST:STUDY_SUMMARY pregnancy induced by common allergen house dust mite (HDM) was used to ST:STUDY_SUMMARY investigate changes in lipid composition in amniotic fluid and bronchoalveolar ST:STUDY_SUMMARY lavage fluid (BALF) by ultra-high performance liquid chromatography/tandem mass ST:STUDY_SUMMARY spectrometry (UPLC-MS/MS), revealing the impact of maternal asthma during ST:STUDY_SUMMARY pregnancy on fetal lipid metabolism. In this study, maternal asthma aggravated ST:STUDY_SUMMARY inflammatory indicators and pathological manifestations after fetal allergen ST:STUDY_SUMMARY exposure, creating a high oxidative stress growth environment for the fetus, and ST:STUDY_SUMMARY causing metabolic differences in various lipid groups, including ST:STUDY_SUMMARY phosphatidylethanolamine (PE), phosphatidylglycerol (PG), and fatty acids (FA), ST:STUDY_SUMMARY indicating significant lipid metabolism disorders. Improving lipid metabolism ST:STUDY_SUMMARY may help asthmatic pregnant women maintain healthy fetal development. ST:INSTITUTE Nanjing University of Chinese Medicine ST:LAST_NAME Fang ST:FIRST_NAME Huafeng ST:ADDRESS No.138 xianlin road, nanjing city, Nanjing, China, 210046, China ST:EMAIL Riorofhf@outlook.com ST:PHONE +86 18852416998 #SUBJECT SU:SUBJECT_TYPE Mammal SU:SUBJECT_SPECIES Rattus norvegicus SU:TAXONOMY_ID 10116 #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 P1 PBS1 Factor:Control type=Amniotic fluid; Genotype=Wild-type; RAW_FILE_NAME=P1.RAW SUBJECT_SAMPLE_FACTORS P2 PBS2 Factor:Control type=Amniotic fluid; Genotype=Wild-type; RAW_FILE_NAME=P2.RAW SUBJECT_SAMPLE_FACTORS P3 PBS3 Factor:Control type=Amniotic fluid; Genotype=Wild-type; RAW_FILE_NAME=P3.RAW SUBJECT_SAMPLE_FACTORS P4 PBS4 Factor:Control type=Amniotic fluid; Genotype=Wild-type; RAW_FILE_NAME=P4.RAW SUBJECT_SAMPLE_FACTORS P6 PBS6 Factor:Control type=Amniotic fluid; Genotype=Wild-type; RAW_FILE_NAME=P6.RAW SUBJECT_SAMPLE_FACTORS P7 PBS7 Factor:Control type=Amniotic fluid; Genotype=Wild-type; RAW_FILE_NAME=P7.RAW SUBJECT_SAMPLE_FACTORS M3 HDM3 Factor:HDM type=Amniotic fluid; Genotype=Wild-type; RAW_FILE_NAME=M3.RAW SUBJECT_SAMPLE_FACTORS M4 HDM4 Factor:HDM type=Amniotic fluid; Genotype=Wild-type; RAW_FILE_NAME=M4.RAW SUBJECT_SAMPLE_FACTORS M5 HDM5 Factor:HDM type=Amniotic fluid; Genotype=Wild-type; RAW_FILE_NAME=M5.RAW SUBJECT_SAMPLE_FACTORS M6 HDM6 Factor:HDM type=Amniotic fluid; Genotype=Wild-type; RAW_FILE_NAME=M6.RAW SUBJECT_SAMPLE_FACTORS M7 HDM7 Factor:HDM type=Amniotic fluid; Genotype=Wild-type; RAW_FILE_NAME=M7.RAW SUBJECT_SAMPLE_FACTORS M8 HDM8 Factor:HDM type=Amniotic fluid; Genotype=Wild-type; RAW_FILE_NAME=M8.RAW #COLLECTION CO:COLLECTION_SUMMARY Amniotic fluid was extracted from the amniotic membrane of GD18 gestational rats CO:COLLECTION_SUMMARY and stored at -80℃ CO:SAMPLE_TYPE Amniotic fluid CO:STORAGE_CONDITIONS -80℃ #TREATMENT TR:TREATMENT_SUMMARY In the sensitization group, the sensitization solution was injected TR:TREATMENT_SUMMARY intritoneally every 2 days before pregnancy, with 100ul of sensitization TR:TREATMENT_SUMMARY solution injected each time, equivalent to 20ug of HDM injection, for a total of TR:TREATMENT_SUMMARY 3 times. The control group was given the same amount of PBS injection. Use of TR:TREATMENT_SUMMARY HDM nasal drops during pregnancy #SAMPLEPREP SP:SAMPLEPREP_SUMMARY 500 μL of amniotic fluid was lyophilized and redissolved in 200 μL of SP:SAMPLEPREP_SUMMARY deionized water. Then, 80 μL of redissolved amniotic fluid was pipetted off SP:SAMPLEPREP_SUMMARY into a 1.5 mL centrifuge tube containing 225 μL of ice-cooled methanol (Merck, SP:SAMPLEPREP_SUMMARY Germany) pre-mixed with lyso PE (17:1; LM171LPE-11), SM (17:0; 170SM-13), and PE SP:SAMPLEPREP_SUMMARY (17:0/17:0; LM170PE-19) internal standards (5 μg·mL-1; Avanti Polar Lipids, SP:SAMPLEPREP_SUMMARY USA). The solution was vortexed for 10 s and added with 750 μL of ice-cooled SP:SAMPLEPREP_SUMMARY MTBE. The mixture was shaken for 10 min at 4 °C, added with 188 μL of SP:SAMPLEPREP_SUMMARY deionized water, vortexed for 20 s, and centrifuged at 18000 rpm for 2 min at 4 SP:SAMPLEPREP_SUMMARY °C. 350 μL of the upper layer (the organic phase, mainly including lipids) and SP:SAMPLEPREP_SUMMARY 110 μL of the bottom layer (the aqueous phase, mainly including polar SP:SAMPLEPREP_SUMMARY substances) were separately transferred to a new centrifuge tube (1.5 mL). The SP:SAMPLEPREP_SUMMARY samples were dried using the Savant SPD1010 vacuum centrifugal concentrator SP:SAMPLEPREP_SUMMARY (Thermo Fisher Scientific, USA) and stored at -20 °C before testing. Lipids in SP:SAMPLEPREP_SUMMARY the upper layers were lysed with 110 μL of methanol-toluene (9:1) solution. #CHROMATOGRAPHY CH:CHROMATOGRAPHY_TYPE Reversed phase CH:INSTRUMENT_NAME Thermo UltiMate 3000 RS CH:COLUMN_NAME Waters Acquity UPLC CSH C18 column (100 mm×2.1mm, 1.7 m) CH:SOLVENT_A 60％acetonitrile；40％water CH:SOLVENT_B 90％isopropanol；10％ acetonitrile；0.1% formic acid；10mM ammonium formate CH:FLOW_GRADIENT 15% B at 0 min, 30% B at 0–2 min, 48% B at 2–2.5 min, 82% B at 2.5–11 min, CH:FLOW_GRADIENT 99% B at 11–12 min and 15% B at 12–15 min. CH:FLOW_RATE 0.6 mL/min CH:COLUMN_TEMPERATURE 65℃ #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:INSTRUMENT_NAME Thermo Q Exactive Orbitrap MS:INSTRUMENT_TYPE Other MS:MS_TYPE ESI MS:ION_MODE NEGATIVE MS:MS_COMMENTS Q Exactive Hybrid Quadrupole-Orbitrap Mass Spectrometer was used for both MS:MS_COMMENTS positive and negative ion modes. Parameters of mass spectrometry: spray voltage MS:MS_COMMENTS was 3.5 kV (positive) and 3.0 kV (negative); for both ionization modes, sheath MS:MS_COMMENTS gas, aux gas, capillary temperature, and heater temperature were maintained at MS:MS_COMMENTS 35 arb, 15 arb, 325 °C and 300 °C, respectively; scan range was m/z MS:MS_COMMENTS 215–1800. MS:MS_RESULTS_FILE ST002765_AN004501_Results.txt UNITS:Peak area Has m/z:Yes Has RT:Yes RT units:Minutes #END