#METABOLOMICS WORKBENCH rioro_20230630_054058 DATATRACK_ID:4131 STUDY_ID:ST002799 ANALYSIS_ID:AN004555 PROJECT_ID:PR001746
VERSION             	1
CREATED_ON             	July 28, 2023, 8:55 am
#PROJECT
PR:PROJECT_TITLE                 	Gestational asthma causes lipid disturbance in fetal alveolar lavage fluid
PR:PROJECT_SUMMARY               	Gestational asthma leads to lipid disturbance of fetal alveolar lavage fluid and
PR:PROJECT_SUMMARY               	lung development impairment
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                   	Gestational asthma causes lipid disturbance in fetal alveolar lavage fluid
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           	PP1	PBS+PBS1	Control:Control	Alveolar lavage fluid=Alveolar lavage fluid; Wild-type=Wild-type; RAW_FILE_NAME=PP1.RAW
SUBJECT_SAMPLE_FACTORS           	PP3	PBS+PBS3	Control:Control	Alveolar lavage fluid=Alveolar lavage fluid; Wild-type=Wild-type; RAW_FILE_NAME=PP3.RAW
SUBJECT_SAMPLE_FACTORS           	PP4	PBS+PBS4	Control:Control	Alveolar lavage fluid=Alveolar lavage fluid; Wild-type=Wild-type; RAW_FILE_NAME=PP4.RAW
SUBJECT_SAMPLE_FACTORS           	PP5	PBS+PBS5	Control:Control	Alveolar lavage fluid=Alveolar lavage fluid; Wild-type=Wild-type; RAW_FILE_NAME=PP5.RAW
SUBJECT_SAMPLE_FACTORS           	PP6	PBS+PBS6	Control:Control	Alveolar lavage fluid=Alveolar lavage fluid; Wild-type=Wild-type; RAW_FILE_NAME=PP6.RAW
SUBJECT_SAMPLE_FACTORS           	PP7	PBS+PBS7	Control:Control	Alveolar lavage fluid=Alveolar lavage fluid; Wild-type=Wild-type; RAW_FILE_NAME=PP7.RAW
SUBJECT_SAMPLE_FACTORS           	PP8	PBS+PBS8	Control:Control	Alveolar lavage fluid=Alveolar lavage fluid; Wild-type=Wild-type; RAW_FILE_NAME=PP8.RAW
SUBJECT_SAMPLE_FACTORS           	PP10	PBS+PBS10	Control:Control	Alveolar lavage fluid=Alveolar lavage fluid; Wild-type=Wild-type; RAW_FILE_NAME=PP10.RAW
SUBJECT_SAMPLE_FACTORS           	ZP1	HDM+PBS1	Control:HDM	Alveolar lavage fluid=Alveolar lavage fluid; Wild-type=Wild-type; RAW_FILE_NAME=ZP1.RAW
SUBJECT_SAMPLE_FACTORS           	ZP3	HDM+PBS3	Control:HDM	Alveolar lavage fluid=Alveolar lavage fluid; Wild-type=Wild-type; RAW_FILE_NAME=ZP3.RAW
SUBJECT_SAMPLE_FACTORS           	ZP5	HDM+PBS5	Control:HDM	Alveolar lavage fluid=Alveolar lavage fluid; Wild-type=Wild-type; RAW_FILE_NAME=ZP5.RAW
SUBJECT_SAMPLE_FACTORS           	ZP6	HDM+PBS6	Control:HDM	Alveolar lavage fluid=Alveolar lavage fluid; Wild-type=Wild-type; RAW_FILE_NAME=ZP6.RAW
SUBJECT_SAMPLE_FACTORS           	ZP7	HDM+PBS7	Control:HDM	Alveolar lavage fluid=Alveolar lavage fluid; Wild-type=Wild-type; RAW_FILE_NAME=ZP7.RAW
SUBJECT_SAMPLE_FACTORS           	ZP8	HDM+PBS8	Control:HDM	Alveolar lavage fluid=Alveolar lavage fluid; Wild-type=Wild-type; RAW_FILE_NAME=ZP8.RAW
SUBJECT_SAMPLE_FACTORS           	ZP9	HDM+PBS9	Control:HDM	Alveolar lavage fluid=Alveolar lavage fluid; Wild-type=Wild-type; RAW_FILE_NAME=ZP9.RAW
SUBJECT_SAMPLE_FACTORS           	ZP10	HDM+PBS10	Control:HDM	Alveolar lavage fluid=Alveolar lavage fluid; Wild-type=Wild-type; RAW_FILE_NAME=ZP10.RAW
SUBJECT_SAMPLE_FACTORS           	ZH1	HDM+HDM1	Control:HDM	Alveolar lavage fluid=Alveolar lavage fluid; Wild-type=Wild-type; RAW_FILE_NAME=ZH1.RAW
SUBJECT_SAMPLE_FACTORS           	ZH2	HDM+HDM2	Control:HDM	Alveolar lavage fluid=Alveolar lavage fluid; Wild-type=Wild-type; RAW_FILE_NAME=ZH2.RAW
SUBJECT_SAMPLE_FACTORS           	ZH3	HDM+HDM3	Control:HDM	Alveolar lavage fluid=Alveolar lavage fluid; Wild-type=Wild-type; RAW_FILE_NAME=ZH3.RAW
SUBJECT_SAMPLE_FACTORS           	ZH4	HDM+HDM4	Control:HDM	Alveolar lavage fluid=Alveolar lavage fluid; Wild-type=Wild-type; RAW_FILE_NAME=ZH4.RAW
SUBJECT_SAMPLE_FACTORS           	ZH6	HDM+HDM6	Control:HDM	Alveolar lavage fluid=Alveolar lavage fluid; Wild-type=Wild-type; RAW_FILE_NAME=ZH6.RAW
SUBJECT_SAMPLE_FACTORS           	ZH7	HDM+HDM7	Control:HDM	Alveolar lavage fluid=Alveolar lavage fluid; Wild-type=Wild-type; RAW_FILE_NAME=ZH7.RAW
SUBJECT_SAMPLE_FACTORS           	ZH8	HDM+HDM8	Control:HDM	Alveolar lavage fluid=Alveolar lavage fluid; Wild-type=Wild-type; RAW_FILE_NAME=ZH8.RAW
SUBJECT_SAMPLE_FACTORS           	ZH10	HDM+HDM10	Control:HDM	Alveolar lavage fluid=Alveolar lavage fluid; Wild-type=Wild-type; RAW_FILE_NAME=ZH10.RAW
#COLLECTION
CO:COLLECTION_SUMMARY            	The mother rats with gestational asthma had a natural pregnancy, and the fetus
CO:COLLECTION_SUMMARY            	was given 1.5mlPBS to extract alveolar lavage fluid on day 28
CO:SAMPLE_TYPE                   	Bronchoalveolar lavage
CO:STORAGE_CONDITIONS            	-80℃
#TREATMENT
TR:TREATMENT_SUMMARY             	The fetuses born to rats with gestational asthma were given HDM nose drops from
TR:TREATMENT_SUMMARY             	day 21 and continued for 5 days, and the same amount of PBS nose drops were used
TR:TREATMENT_SUMMARY             	in blank control
#SAMPLEPREP
SP:SAMPLEPREP_SUMMARY            	500 μL of alveolar lavage fluid was lyophilized and redissolved in 200 μL of
SP:SAMPLEPREP_SUMMARY            	deionized water. Then, 80 μL of redissolved alveolar lavage fluid was pipetted
SP:SAMPLEPREP_SUMMARY            	off into a 1.5 mL centrifuge tube containing 225 μL of ice-cooled methanol
SP:SAMPLEPREP_SUMMARY            	(Merck, Germany) pre-mixed with lyso PE (17:1; LM171LPE-11), SM (17:0;
SP:SAMPLEPREP_SUMMARY            	170SM-13), and PE (17:0/17:0; LM170PE-19) internal standards (5 μg·mL-1;
SP:SAMPLEPREP_SUMMARY            	Avanti Polar Lipids, USA). The solution was vortexed for 10 s and added with 750
SP:SAMPLEPREP_SUMMARY            	μL of ice-cooled MTBE. The mixture was shaken for 10 min at 4 °C, added with
SP:SAMPLEPREP_SUMMARY            	188 μL of deionized water, vortexed for 20 s, and centrifuged at 18000 rpm for
SP:SAMPLEPREP_SUMMARY            	2 min at 4 °C. 350 μL of the upper layer (the organic phase, mainly including
SP:SAMPLEPREP_SUMMARY            	lipids) and 110 μL of the bottom layer (the aqueous phase, mainly including
SP:SAMPLEPREP_SUMMARY            	polar substances) were separately transferred to a new centrifuge tube (1.5 mL).
SP:SAMPLEPREP_SUMMARY            	The 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 Dionex Ultimate 3000 RS
CH:COLUMN_NAME                   	Waters ACQUITY UPLC BEH C18 (100 x 2.1mm,1.7um)
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               	ST002799_AN004555_Results.txt	UNITS:Peak area	Has m/z:Yes	Has RT:Yes	RT units:Minutes
#END