#METABOLOMICS WORKBENCH rioro_20230629_060334 DATATRACK_ID:4126 STUDY_ID:ST002765 ANALYSIS_ID:AN004500 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                      	POSITIVE
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_AN004500_Results.txt	UNITS:Peak area	Has m/z:Yes	Has RT:Yes	RT units:Minutes
#END