#METABOLOMICS WORKBENCH Kathang_20250319_202355 DATATRACK_ID:5749 STUDY_ID:ST003839 ANALYSIS_ID:AN006308 PROJECT_ID:PR002398
VERSION             	1
CREATED_ON             	March 31, 2025, 2:33 pm
#PROJECT
PR:PROJECT_TITLE                 	Taurochenodeoxycholic acid levels are associated with scar hypertrophy in the
PR:PROJECT_TITLE                 	rabbit ear hypertrophic scar model.
PR:PROJECT_SUMMARY               	The aim of this study was to investigate the relationship between gut
PR:PROJECT_SUMMARY               	microbiota, metabolites and scar hypertrophy using the rabbit ear hypertrophic
PR:PROJECT_SUMMARY               	scar model. On postoperative day 28, the rabbits were divided into
PR:PROJECT_SUMMARY               	scar-susceptible (group H) and scar-insusceptible (group L) groups according to
PR:PROJECT_SUMMARY               	macroscopic scar thickness. Using fecal microbiota transplantation (FMT), we
PR:PROJECT_SUMMARY               	exchanged the gut microbiota of group H and group L. Untargeted metabolomics was
PR:PROJECT_SUMMARY               	performed before and after FMT. We found that in the rabbit ear hypertrophic
PR:PROJECT_SUMMARY               	scar model, scar hypertrophy was more severe in group H and was associated with
PR:PROJECT_SUMMARY               	fecal microbiota and their metabolites. Gut microbiota components and key genera
PR:PROJECT_SUMMARY               	were transferred between group H and group L by FMT, accompanied by altered
PR:PROJECT_SUMMARY               	levels of key metabolites such as TCDCA (bile acid taurochenodeoxycholic acid).
PR:PROJECT_SUMMARY               	FMT relieved scar hypertrophy in group H and vice versa. TCDCA were decreased in
PR:PROJECT_SUMMARY               	group H rabbit and its relative abundance was successfully reversed by
PR:PROJECT_SUMMARY               	heterochronic FMT.
PR:INSTITUTE                     	Shenzhen Center for Chronic Disease Control
PR:DEPARTMENT                    	Plastic surgery, Shenzhen Institute of Dermatology
PR:LAST_NAME                     	Liang
PR:FIRST_NAME                    	Zhengyun
PR:ADDRESS                       	No. 2021, Buxin Road, Luohu District, Shenzhen, Guangdong, 51802, China
PR:EMAIL                         	18819481519@163.com
PR:PHONE                         	18819481519
#STUDY
ST:STUDY_TITLE                   	Taurochenodeoxycholic acid levels are associated with scar hypertrophy in the
ST:STUDY_TITLE                   	rabbit ear hypertrophic scar model.
ST:STUDY_SUMMARY                 	New Zealand white rabbits were used to establish the rabbit ear hypertrophic
ST:STUDY_SUMMARY                 	scar model. On postoperative day 28, the rabbits were divided into
ST:STUDY_SUMMARY                 	scar-susceptible (group H) and scar-insusceptible (group L) groups according to
ST:STUDY_SUMMARY                 	macroscopic scar thickness. Using fecal microbiota transplantation (FMT), we
ST:STUDY_SUMMARY                 	exchanged the gut microbiota of group H and group L. Untargeted metabolomics was
ST:STUDY_SUMMARY                 	performed before and after FMT. We found that in the rabbit ear hypertrophic
ST:STUDY_SUMMARY                 	scar model, scar hypertrophy was more severe in group H and was associated with
ST:STUDY_SUMMARY                 	fecal microbiota and their metabolites. Gut microbiota components and key genera
ST:STUDY_SUMMARY                 	were transferred between group H and group L by FMT, accompanied by altered
ST:STUDY_SUMMARY                 	levels of key metabolites such as TCDCA (bile acid taurochenodeoxycholic acid).
ST:STUDY_SUMMARY                 	FMT relieved scar hypertrophy in group H and vice versa. TCDCA were decreased in
ST:STUDY_SUMMARY                 	group H rabbit and its relative abundance was successfully reversed by
ST:STUDY_SUMMARY                 	heterochronic FMT.
ST:INSTITUTE                     	Shenzhen Center for Chronic Disease Control,
ST:DEPARTMENT                    	Plastic surgery, Shenzhen Institute of Dermatology
ST:LAST_NAME                     	Liang
ST:FIRST_NAME                    	Zhengyun
ST:ADDRESS                       	No. 2021, Buxin Road, Luohu District, Shenzhen 518020, China
ST:EMAIL                         	18819481519@163.com
ST:PHONE                         	18819481519
#SUBJECT
SU:SUBJECT_TYPE                  	Mammal
SU:SUBJECT_SPECIES               	Oryctolagus cuniculus
SU:TAXONOMY_ID                   	9986
SU:GENOTYPE_STRAIN               	Wildtype
SU:GENDER                        	Male and female
#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           	A05_T1	A05_T1	Treatment:H | Time point:T1 | Sample source:feces	RAW_FILE_NAME=HFX12_CN1_FZTM230233451-1A.raw
SUBJECT_SAMPLE_FACTORS           	A29_T1	A29_T1	Treatment:H | Time point:T1 | Sample source:feces	RAW_FILE_NAME=HFX12_CN1_FZTM230233452-1A.raw
SUBJECT_SAMPLE_FACTORS           	A25_T1	A25_T1	Treatment:H | Time point:T1 | Sample source:feces	RAW_FILE_NAME=HFX12_CN1_FZTM230233453-1A.raw
SUBJECT_SAMPLE_FACTORS           	A46_T1	A46_T1	Treatment:H | Time point:T1 | Sample source:feces	RAW_FILE_NAME=HFX12_CN1_FZTM230233454-1A.raw
SUBJECT_SAMPLE_FACTORS           	A33_T1	A33_T1	Treatment:H | Time point:T1 | Sample source:feces	RAW_FILE_NAME=HFX12_CN1_FZTM230233455-1A.raw
SUBJECT_SAMPLE_FACTORS           	A38_T1	A38_T1	Treatment:H | Time point:T1 | Sample source:feces	RAW_FILE_NAME=HFX12_CN1_FZTM230233457-1A.raw
SUBJECT_SAMPLE_FACTORS           	A07_T1	A07_T1	Treatment:H | Time point:T1 | Sample source:feces	RAW_FILE_NAME=HFX12_CN1_FZTM230233458-1A.raw
SUBJECT_SAMPLE_FACTORS           	A30_T1	A30_T1	Treatment:H | Time point:T1 | Sample source:feces	RAW_FILE_NAME=HFX12_CN1_FZTM230233460-1A.raw
SUBJECT_SAMPLE_FACTORS           	A09_T1	A09_T1	Treatment:H | Time point:T1 | Sample source:feces	RAW_FILE_NAME=HFX12_CN1_FZTM230233462-1A.raw
SUBJECT_SAMPLE_FACTORS           	A48_T1	A48_T1	Treatment:H | Time point:T1 | Sample source:feces	RAW_FILE_NAME=HFX12_CN1_FZTM230233463-1A.raw
SUBJECT_SAMPLE_FACTORS           	A33_T4	A33_T4	Treatment:H-FMT | Time point:T4 | Sample source:feces	RAW_FILE_NAME=HFX12_CN1_FZTM230233469-1A.raw
SUBJECT_SAMPLE_FACTORS           	A38_T4	A38_T4	Treatment:H-FMT | Time point:T4 | Sample source:feces	RAW_FILE_NAME=HFX12_CN1_FZTM230233470-1A.raw
SUBJECT_SAMPLE_FACTORS           	A07_T4	A07_T4	Treatment:H-FMT | Time point:T4 | Sample source:feces	RAW_FILE_NAME=HFX12_CN1_FZTM230233471-1A.raw
SUBJECT_SAMPLE_FACTORS           	A30_T4	A30_T4	Treatment:H-FMT | Time point:T4 | Sample source:feces	RAW_FILE_NAME=HFX12_CN1_FZTM230233472-1A.raw
SUBJECT_SAMPLE_FACTORS           	A09_T4	A09_T4	Treatment:H-FMT | Time point:T4 | Sample source:feces	RAW_FILE_NAME=HFX12_CN1_FZTM230233473-1A.raw
SUBJECT_SAMPLE_FACTORS           	A48_T4	A48_T4	Treatment:H-FMT | Time point:T4 | Sample source:feces	RAW_FILE_NAME=HFX12_CN1_FZTM230233474-1A.raw
SUBJECT_SAMPLE_FACTORS           	A06_T1	A06_T1	Treatment:L | Time point:T1 | Sample source:feces	RAW_FILE_NAME=HFX12_CN1_FZTM230233548-1A.raw
SUBJECT_SAMPLE_FACTORS           	A08_T1	A08_T1	Treatment:L | Time point:T1 | Sample source:feces	RAW_FILE_NAME=HFX12_CN1_FZTM230233549-1A.raw
SUBJECT_SAMPLE_FACTORS           	A42_T1	A42_T1	Treatment:L | Time point:T1 | Sample source:feces	RAW_FILE_NAME=HFX12_CN1_FZTM230233550-1A.raw
SUBJECT_SAMPLE_FACTORS           	A44_T1	A44_T1	Treatment:L | Time point:T1 | Sample source:feces	RAW_FILE_NAME=HFX12_CN1_FZTM230233551-1A.raw
SUBJECT_SAMPLE_FACTORS           	A34_T1	A34_T1	Treatment:L | Time point:T1 | Sample source:feces	RAW_FILE_NAME=HFX12_CN1_FZTM230233556-1A.raw
SUBJECT_SAMPLE_FACTORS           	A39_T1	A39_T1	Treatment:L | Time point:T1 | Sample source:feces	RAW_FILE_NAME=HFX12_CN1_FZTM230233557-1A.raw
SUBJECT_SAMPLE_FACTORS           	A40_T1	A40_T1	Treatment:L | Time point:T1 | Sample source:feces	RAW_FILE_NAME=HFX12_CN1_FZTM230233558-1A.raw
SUBJECT_SAMPLE_FACTORS           	A03_T1	A03_T1	Treatment:L | Time point:T1 | Sample source:feces	RAW_FILE_NAME=HFX12_CN1_FZTM230233559-1A.raw
SUBJECT_SAMPLE_FACTORS           	A31_T1	A31_T1	Treatment:L | Time point:T1 | Sample source:feces	RAW_FILE_NAME=HFX12_CN1_FZTM230233560-1A.raw
SUBJECT_SAMPLE_FACTORS           	A36_T1	A36_T1	Treatment:L | Time point:T1 | Sample source:feces	RAW_FILE_NAME=HFX12_CN1_FZTM230233561-1A.raw
SUBJECT_SAMPLE_FACTORS           	A34_T4	A34_T4	Treatment:L-FMT | Time point:T4 | Sample source:feces	RAW_FILE_NAME=HFX12_CN1_FZTM230233570-1A.raw
SUBJECT_SAMPLE_FACTORS           	A39_T4	A39_T4	Treatment:L-FMT | Time point:T4 | Sample source:feces	RAW_FILE_NAME=HFX12_CN1_FZTM230233572-1A.raw
SUBJECT_SAMPLE_FACTORS           	A40_T4	A40_T4	Treatment:L-FMT | Time point:T4 | Sample source:feces	RAW_FILE_NAME=HFX12_CN1_FZTM230233573-1A.raw
SUBJECT_SAMPLE_FACTORS           	A03_T4	A03_T4	Treatment:L-FMT | Time point:T4 | Sample source:feces	RAW_FILE_NAME=HFX12_CN1_FZTM230233575-1A.raw
SUBJECT_SAMPLE_FACTORS           	A31_T4	A31_T4	Treatment:L-FMT | Time point:T4 | Sample source:feces	RAW_FILE_NAME=HFX12_CN1_FZTM230233577-1A.raw
SUBJECT_SAMPLE_FACTORS           	A36_T4	A36_T4	Treatment:L-FMT | Time point:T4 | Sample source:feces	RAW_FILE_NAME=HFX12_CN1_FZTM230233578-1A.raw
#COLLECTION
CO:COLLECTION_SUMMARY            	Four full-thickness 1.5 cm in diameter round wounds were created on the ventral
CO:COLLECTION_SUMMARY            	side of each rabbit ear using sterile surgical blades following iodophor
CO:COLLECTION_SUMMARY            	sterilization. Sterile boxes, pads, tweezers and fecal containers were utilized
CO:COLLECTION_SUMMARY            	for rabbit fecal pellets collection. Fecal pellets were collected at
CO:COLLECTION_SUMMARY            	“grouping” (postoperative day 28, T1) and “end” (postoperative day 63,
CO:COLLECTION_SUMMARY            	T4) time points.
CO:SAMPLE_TYPE                   	Feces
#TREATMENT
TR:TREATMENT_SUMMARY             	Macroscopic scar thickness (mm) was measured weekly from postoperative day 21
TR:TREATMENT_SUMMARY             	using a vernier caliper. On postoperative day 28, rabbits were grouped by
TR:TREATMENT_SUMMARY             	macroscopic scar thickness. Rabbits with MST values greater than 1.9 mm were
TR:TREATMENT_SUMMARY             	assigned to the high-risk/susceptible group for hypertrophic scars (group H),
TR:TREATMENT_SUMMARY             	otherwise, they were assigned to the low-risk/insusceptible group (group L).
TR:TREATMENT_SUMMARY             	Using fecal microbiota transplantation (FMT), we exchanged the gut microbiota of
TR:TREATMENT_SUMMARY             	group H and group L.
#SAMPLEPREP
SP:SAMPLEPREP_SUMMARY            	After being grounded with liquid nitrogen, 100 mg rabbit fecal samples were
SP:SAMPLEPREP_SUMMARY            	added to 500 μL 80% methanol-water solution. After dilution and centrifugation,
SP:SAMPLEPREP_SUMMARY            	the supernatant with 53% methanol was collected for the liquid
SP:SAMPLEPREP_SUMMARY            	chromatography–mass spectrometry (LC-MS) analysis.
#CHROMATOGRAPHY
CH:CHROMATOGRAPHY_SUMMARY        	Samples were injected onto a Hypesil Gold column (100×2.1 mm, 1.9μm) using a
CH:CHROMATOGRAPHY_SUMMARY        	12-min linear gradient at a flow rate of 0.2mL/min.Solvent A was 5 mM ammonium
CH:CHROMATOGRAPHY_SUMMARY        	acetate and olvent B was Methanol.Solvent A was maintained at pH 9.0.The
CH:CHROMATOGRAPHY_SUMMARY        	gradient was set as follows:2% B,0-1.5 min; 2-85% B,1.5-3 min; 85-100% B,3-10
CH:CHROMATOGRAPHY_SUMMARY        	min；100-2% B, 10-10.1min ；2% B,10.1-12 min.
CH:CHROMATOGRAPHY_TYPE           	HILIC
CH:INSTRUMENT_NAME               	Thermo Vanquish
CH:COLUMN_NAME                   	Thermo Hypersil GOLD aQ (100 x 2.1mm,1.9um)
CH:SOLVENT_A                     	100% water; 5 mM ammonium acetate
CH:SOLVENT_B                     	100% Methanol
CH:FLOW_GRADIENT                 	2% B,0-1.5 min; 2-85% B,1.5-3 min; 85-100% B,3-10 min；100-2% B, 10-10.1min
CH:FLOW_GRADIENT                 	；2% B,10.1-12 min.
CH:FLOW_RATE                     	0.2 mL/min
CH:COLUMN_TEMPERATURE            	40
#ANALYSIS
AN:ANALYSIS_TYPE                 	MS
#MS
MS:INSTRUMENT_NAME               	Thermo Q Exactive HF-X Orbitrap
MS:INSTRUMENT_TYPE               	Orbitrap
MS:MS_TYPE                       	Other
MS:ION_MODE                      	NEGATIVE
MS:MS_COMMENTS                   	The Compound Discoverer 3.1 (CD3.1, ThermoFisher) was used for peak alignment,
MS:MS_COMMENTS                   	selection and normalization of raw data based on retention time, signal/noise
MS:MS_COMMENTS                   	ratio and other parameters.
MS:MS_RESULTS_FILE               	ST003839_AN006308_Results.txt	UNITS:peak area	Has m/z:Yes	Has RT:Yes	RT units:Minutes
#END