#METABOLOMICS WORKBENCH zhouyiyang_20250221_183530 DATATRACK_ID:5665 STUDY_ID:ST003767 ANALYSIS_ID:AN006184 PROJECT_ID:PR002350
VERSION             	1
CREATED_ON             	February 24, 2025, 4:44 pm
#PROJECT
PR:PROJECT_TITLE                 	N-acetylneuraminic Acid may play Links Secondary Neurodegeneration and Cognitive
PR:PROJECT_TITLE                 	Impairment in Cortical Photothrombotic Stroke Mouse Model
PR:PROJECT_SUMMARY               	ABSTRACT Background: Post-stroke cognitive impairment (PSCI) is a major sequelae
PR:PROJECT_SUMMARY               	of ischemic stroke (IS), but the associated mechanisms remain unclear. Recent
PR:PROJECT_SUMMARY               	studies have shown that cortical stroke can cause distal brain disturbances,
PR:PROJECT_SUMMARY               	leading to cognitive decline. This study aims to investigate the
PR:PROJECT_SUMMARY               	neuropathological and metabolic changes over time in the remote brain region
PR:PROJECT_SUMMARY               	associated with cognitive impairment after cortical stroke. Methods: We assessed
PR:PROJECT_SUMMARY               	cognitive function in photothrombotic (PT) mice for 84 days and identified
PR:PROJECT_SUMMARY               	distal brain regions with secondary neurodegeneration (SND) using voxel-based
PR:PROJECT_SUMMARY               	morphometry (VBM). Integrated untargeted and targeted metabolomics method was
PR:PROJECT_SUMMARY               	established to comprehensively analyze the molecular mechanism of SND in this
PR:PROJECT_SUMMARY               	brain region and screen the potential predictive biomarker of PSCI. Furthermore,
PR:PROJECT_SUMMARY               	the mechanism of the biomarker with PSCI and SND was studied in vitro. Results:
PR:PROJECT_SUMMARY               	The results show that recent memory impairment, remote dysfunction, and anxiety
PR:PROJECT_SUMMARY               	persisted for 84 days in PT mice. The hippocampus of cognitively impaired PT
PR:PROJECT_SUMMARY               	mice developed SND and metabolic disorders, particularly oxidative stress, lipid
PR:PROJECT_SUMMARY               	peroxidation, and inflammation. N-acetylneuramic acid (Neu5Ac) was screened in
PR:PROJECT_SUMMARY               	the hippocampus of mice, serum of mice and 154 IS patients. Neu5Ac promotes
PR:PROJECT_SUMMARY               	oxidative stress and inflammation in microglia. Conclusions: Our results suggest
PR:PROJECT_SUMMARY               	that hippocampal SND is closely related to cognitive impairment in PT mice,
PR:PROJECT_SUMMARY               	while oxidative stress and inflammation are important factors in hippocampal
PR:PROJECT_SUMMARY               	SND. Neu5Ac may aggravate PSCI by causing hippocampal SND through inducting of
PR:PROJECT_SUMMARY               	microglia-driven oxidative stress and inflammation, and it may thus be a
PR:PROJECT_SUMMARY               	potential predictive biomarker of PSCI.
PR:INSTITUTE                     	Wenzhou Medical University
PR:LAST_NAME                     	Zhou
PR:FIRST_NAME                    	Yiyang
PR:ADDRESS                       	University Town, Chashan, Wenzhou, Zhejiang, 325035 P.R China
PR:EMAIL                         	zhouyiyang@wmu.edu.cn
PR:PHONE                         	+8613806831161
#STUDY
ST:STUDY_TITLE                   	N-acetylneuraminic Acid may play Links Secondary Neurodegeneration and Cognitive
ST:STUDY_TITLE                   	Impairment in Cortical Photothrombotic Stroke Mouse Model
ST:STUDY_SUMMARY                 	ABSTRACT Background: Post-stroke cognitive impairment (PSCI) is a major sequelae
ST:STUDY_SUMMARY                 	of ischemic stroke (IS), but the associated mechanisms remain unclear. Recent
ST:STUDY_SUMMARY                 	studies have shown that cortical stroke can cause distal brain disturbances,
ST:STUDY_SUMMARY                 	leading to cognitive decline. This study aims to investigate the
ST:STUDY_SUMMARY                 	neuropathological and metabolic changes over time in the remote brain region
ST:STUDY_SUMMARY                 	associated with cognitive impairment after cortical stroke. Methods: We assessed
ST:STUDY_SUMMARY                 	cognitive function in photothrombotic (PT) mice for 84 days and identified
ST:STUDY_SUMMARY                 	distal brain regions with secondary neurodegeneration (SND) using voxel-based
ST:STUDY_SUMMARY                 	morphometry (VBM). Integrated untargeted and targeted metabolomics method was
ST:STUDY_SUMMARY                 	established to comprehensively analyze the molecular mechanism of SND in this
ST:STUDY_SUMMARY                 	brain region and screen the potential predictive biomarker of PSCI. Furthermore,
ST:STUDY_SUMMARY                 	the mechanism of the biomarker with PSCI and SND was studied in vitro. Results:
ST:STUDY_SUMMARY                 	The results show that recent memory impairment, remote dysfunction, and anxiety
ST:STUDY_SUMMARY                 	persisted for 84 days in PT mice. The hippocampus of cognitively impaired PT
ST:STUDY_SUMMARY                 	mice developed SND and metabolic disorders, particularly oxidative stress, lipid
ST:STUDY_SUMMARY                 	peroxidation, and inflammation. N-acetylneuramic acid (Neu5Ac) was screened in
ST:STUDY_SUMMARY                 	the hippocampus of mice, serum of mice and 154 IS patients. Neu5Ac promotes
ST:STUDY_SUMMARY                 	oxidative stress and inflammation in microglia. Conclusions: Our results suggest
ST:STUDY_SUMMARY                 	that hippocampal SND is closely related to cognitive impairment in PT mice,
ST:STUDY_SUMMARY                 	while oxidative stress and inflammation are important factors in hippocampal
ST:STUDY_SUMMARY                 	SND. Neu5Ac may aggravate PSCI by causing hippocampal SND through inducting of
ST:STUDY_SUMMARY                 	microglia-driven oxidative stress and inflammation, and it may thus be a
ST:STUDY_SUMMARY                 	potential predictive biomarker of PSCI
ST:INSTITUTE                     	
ST:LAST_NAME                     	Zhou
ST:FIRST_NAME                    	Yiyang
ST:ADDRESS                       	University Town, Chashan, Wenzhou, Zhejiang, 325035 P.R China
ST:EMAIL                         	zhouyiyang@wmu.edu.cn
ST:PHONE                         	13806831161
#SUBJECT
SU:SUBJECT_TYPE                  	Mammal
SU:SUBJECT_SPECIES               	Mus musculus
SU:TAXONOMY_ID                   	10090
#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           	-	C11	Sample source:hippocampus | Sample_type:C1	RAW_FILE_NAME(Positive_mzXML_FILE_NAME)=C11-LHPOS.mzXML; RAW_FILE_NAME(Negative_mzXML_FILE_NAME)=C11-LHneg.mzXML
SUBJECT_SAMPLE_FACTORS           	-	C12	Sample source:hippocampus | Sample_type:C1	RAW_FILE_NAME(Positive_mzXML_FILE_NAME)=C12-LHPOS.mzXML; RAW_FILE_NAME(Negative_mzXML_FILE_NAME)=C12-LHneg.mzXML
SUBJECT_SAMPLE_FACTORS           	-	C13	Sample source:hippocampus | Sample_type:C1	RAW_FILE_NAME(Positive_mzXML_FILE_NAME)=C13-LHPOS.mzXML; RAW_FILE_NAME(Negative_mzXML_FILE_NAME)=C13-LHneg.mzXML
SUBJECT_SAMPLE_FACTORS           	-	C14	Sample source:hippocampus | Sample_type:C1	RAW_FILE_NAME(Positive_mzXML_FILE_NAME)=C14-LHPOS.mzXML; RAW_FILE_NAME(Negative_mzXML_FILE_NAME)=C14-LHneg.mzXML
SUBJECT_SAMPLE_FACTORS           	-	C15	Sample source:hippocampus | Sample_type:C1	RAW_FILE_NAME(Positive_mzXML_FILE_NAME)=C15-LHPOS.mzXML; RAW_FILE_NAME(Negative_mzXML_FILE_NAME)=C15-LHneg.mzXML
SUBJECT_SAMPLE_FACTORS           	-	C16	Sample source:hippocampus | Sample_type:C1	RAW_FILE_NAME(Positive_mzXML_FILE_NAME)=C16-LHPOS.mzXML; RAW_FILE_NAME(Negative_mzXML_FILE_NAME)=C16-LHneg.mzXML
SUBJECT_SAMPLE_FACTORS           	-	C17	Sample source:hippocampus | Sample_type:C1	RAW_FILE_NAME(Positive_mzXML_FILE_NAME)=C17-LHPOS.mzXML; RAW_FILE_NAME(Negative_mzXML_FILE_NAME)=C17-LHneg.mzXML
SUBJECT_SAMPLE_FACTORS           	-	C19	Sample source:hippocampus | Sample_type:C1	RAW_FILE_NAME(Positive_mzXML_FILE_NAME)=C19-LHPOS.mzXML; RAW_FILE_NAME(Negative_mzXML_FILE_NAME)=C19-LHneg.mzXML
SUBJECT_SAMPLE_FACTORS           	-	C21	Sample source:hippocampus | Sample_type:C2	RAW_FILE_NAME(Positive_mzXML_FILE_NAME)=C21-LHPOS.mzXML; RAW_FILE_NAME(Negative_mzXML_FILE_NAME)=C21-LHneg.mzXML
SUBJECT_SAMPLE_FACTORS           	-	C22	Sample source:hippocampus | Sample_type:C2	RAW_FILE_NAME(Positive_mzXML_FILE_NAME)=C22-LHPOS.mzXML; RAW_FILE_NAME(Negative_mzXML_FILE_NAME)=C22-LHneg.mzXML
SUBJECT_SAMPLE_FACTORS           	-	C23	Sample source:hippocampus | Sample_type:C2	RAW_FILE_NAME(Positive_mzXML_FILE_NAME)=C23-LHPOS.mzXML; RAW_FILE_NAME(Negative_mzXML_FILE_NAME)=C23-LHneg.mzXML
SUBJECT_SAMPLE_FACTORS           	-	C24	Sample source:hippocampus | Sample_type:C2	RAW_FILE_NAME(Positive_mzXML_FILE_NAME)=C24-LHPOS.mzXML; RAW_FILE_NAME(Negative_mzXML_FILE_NAME)=C24-LHneg.mzXML
SUBJECT_SAMPLE_FACTORS           	-	C25	Sample source:hippocampus | Sample_type:C2	RAW_FILE_NAME(Positive_mzXML_FILE_NAME)=C25-LHPOS.mzXML; RAW_FILE_NAME(Negative_mzXML_FILE_NAME)=C25-LHneg.mzXML
SUBJECT_SAMPLE_FACTORS           	-	C26	Sample source:hippocampus | Sample_type:C2	RAW_FILE_NAME(Positive_mzXML_FILE_NAME)=C26-LHPOS.mzXML; RAW_FILE_NAME(Negative_mzXML_FILE_NAME)=C26-LHneg.mzXML
SUBJECT_SAMPLE_FACTORS           	-	C27	Sample source:hippocampus | Sample_type:C2	RAW_FILE_NAME(Positive_mzXML_FILE_NAME)=C27-LHPOS.mzXML; RAW_FILE_NAME(Negative_mzXML_FILE_NAME)=C27-LHneg.mzXML
SUBJECT_SAMPLE_FACTORS           	-	C28	Sample source:hippocampus | Sample_type:C2	RAW_FILE_NAME(Positive_mzXML_FILE_NAME)=C28-LHPOS.mzXML; RAW_FILE_NAME(Negative_mzXML_FILE_NAME)=C28-LHneg.mzXML
SUBJECT_SAMPLE_FACTORS           	-	C31	Sample source:hippocampus | Sample_type:C3	RAW_FILE_NAME(Positive_mzXML_FILE_NAME)=C31-LHPOS.mzXML; RAW_FILE_NAME(Negative_mzXML_FILE_NAME)=C31-LHneg.mzXML
SUBJECT_SAMPLE_FACTORS           	-	C32	Sample source:hippocampus | Sample_type:C3	RAW_FILE_NAME(Positive_mzXML_FILE_NAME)=C32-LHPOS.mzXML; RAW_FILE_NAME(Negative_mzXML_FILE_NAME)=C32-LHneg.mzXML
SUBJECT_SAMPLE_FACTORS           	-	C33	Sample source:hippocampus | Sample_type:C3	RAW_FILE_NAME(Positive_mzXML_FILE_NAME)=C33-LHPOS.mzXML; RAW_FILE_NAME(Negative_mzXML_FILE_NAME)=C33-LHneg.mzXML
SUBJECT_SAMPLE_FACTORS           	-	C35	Sample source:hippocampus | Sample_type:C3	RAW_FILE_NAME(Positive_mzXML_FILE_NAME)=C35-LHPOS.mzXML; RAW_FILE_NAME(Negative_mzXML_FILE_NAME)=C35-LHneg.mzXML
SUBJECT_SAMPLE_FACTORS           	-	C36	Sample source:hippocampus | Sample_type:C3	RAW_FILE_NAME(Positive_mzXML_FILE_NAME)=C36-LHPOS.mzXML; RAW_FILE_NAME(Negative_mzXML_FILE_NAME)=C36-LHneg.mzXML
SUBJECT_SAMPLE_FACTORS           	-	C37	Sample source:hippocampus | Sample_type:C3	RAW_FILE_NAME(Positive_mzXML_FILE_NAME)=C37-LHPOS.mzXML; RAW_FILE_NAME(Negative_mzXML_FILE_NAME)=C37-LHneg.mzXML
SUBJECT_SAMPLE_FACTORS           	-	C38	Sample source:hippocampus | Sample_type:C3	RAW_FILE_NAME(Positive_mzXML_FILE_NAME)=C38-LHPOS.mzXML; RAW_FILE_NAME(Negative_mzXML_FILE_NAME)=C38-LHneg.mzXML
SUBJECT_SAMPLE_FACTORS           	-	C39	Sample source:hippocampus | Sample_type:C3	RAW_FILE_NAME(Positive_mzXML_FILE_NAME)=C39-LHPOS.mzXML; RAW_FILE_NAME(Negative_mzXML_FILE_NAME)=C39-LHneg.mzXML
SUBJECT_SAMPLE_FACTORS           	-	M11	Sample source:hippocampus | Sample_type:M1	RAW_FILE_NAME(Positive_mzXML_FILE_NAME)=M11-LHPOS.mzXML; RAW_FILE_NAME(Negative_mzXML_FILE_NAME)=M11-LHneg.mzXML
SUBJECT_SAMPLE_FACTORS           	-	M12	Sample source:hippocampus | Sample_type:M1	RAW_FILE_NAME(Positive_mzXML_FILE_NAME)=M12-LHPOS.mzXML; RAW_FILE_NAME(Negative_mzXML_FILE_NAME)=M12-LHneg.mzXML
SUBJECT_SAMPLE_FACTORS           	-	M13	Sample source:hippocampus | Sample_type:M1	RAW_FILE_NAME(Positive_mzXML_FILE_NAME)=M13-LHPOS.mzXML; RAW_FILE_NAME(Negative_mzXML_FILE_NAME)=M13-LHneg.mzXML
SUBJECT_SAMPLE_FACTORS           	-	M14	Sample source:hippocampus | Sample_type:M1	RAW_FILE_NAME(Positive_mzXML_FILE_NAME)=M14-LHPOS.mzXML; RAW_FILE_NAME(Negative_mzXML_FILE_NAME)=M14-LHneg.mzXML
SUBJECT_SAMPLE_FACTORS           	-	M15	Sample source:hippocampus | Sample_type:M1	RAW_FILE_NAME(Positive_mzXML_FILE_NAME)=M15-LHPOS.mzXML; RAW_FILE_NAME(Negative_mzXML_FILE_NAME)=M15-LHneg.mzXML
SUBJECT_SAMPLE_FACTORS           	-	M16	Sample source:hippocampus | Sample_type:M1	RAW_FILE_NAME(Positive_mzXML_FILE_NAME)=M16-LHPOS.mzXML; RAW_FILE_NAME(Negative_mzXML_FILE_NAME)=M16-LHneg.mzXML
SUBJECT_SAMPLE_FACTORS           	-	M17	Sample source:hippocampus | Sample_type:M1	RAW_FILE_NAME(Positive_mzXML_FILE_NAME)=M17-LHPOS.mzXML; RAW_FILE_NAME(Negative_mzXML_FILE_NAME)=M17-LHneg.mzXML
SUBJECT_SAMPLE_FACTORS           	-	M18	Sample source:hippocampus | Sample_type:M1	RAW_FILE_NAME(Positive_mzXML_FILE_NAME)=M18-LHPOS.mzXML; RAW_FILE_NAME(Negative_mzXML_FILE_NAME)=M18-LHneg.mzXML
SUBJECT_SAMPLE_FACTORS           	-	M21	Sample source:hippocampus | Sample_type:M2	RAW_FILE_NAME(Positive_mzXML_FILE_NAME)=M21-LHPOS.mzXML; RAW_FILE_NAME(Negative_mzXML_FILE_NAME)=M21-LHneg.mzXML
SUBJECT_SAMPLE_FACTORS           	-	M22	Sample source:hippocampus | Sample_type:M2	RAW_FILE_NAME(Positive_mzXML_FILE_NAME)=M22-LHPOS.mzXML; RAW_FILE_NAME(Negative_mzXML_FILE_NAME)=M22-LHneg.mzXML
SUBJECT_SAMPLE_FACTORS           	-	M23	Sample source:hippocampus | Sample_type:M2	RAW_FILE_NAME(Positive_mzXML_FILE_NAME)=M23-LHPOS.mzXML; RAW_FILE_NAME(Negative_mzXML_FILE_NAME)=M23-LHneg.mzXML
SUBJECT_SAMPLE_FACTORS           	-	M24	Sample source:hippocampus | Sample_type:M2	RAW_FILE_NAME(Positive_mzXML_FILE_NAME)=M24-LHPOS.mzXML; RAW_FILE_NAME(Negative_mzXML_FILE_NAME)=M24-LHneg.mzXML
SUBJECT_SAMPLE_FACTORS           	-	M25	Sample source:hippocampus | Sample_type:M2	RAW_FILE_NAME(Positive_mzXML_FILE_NAME)=M25-LHPOS.mzXML; RAW_FILE_NAME(Negative_mzXML_FILE_NAME)=M25-LHneg.mzXML
SUBJECT_SAMPLE_FACTORS           	-	M26	Sample source:hippocampus | Sample_type:M2	RAW_FILE_NAME(Positive_mzXML_FILE_NAME)=M26-LHPOS.mzXML; RAW_FILE_NAME(Negative_mzXML_FILE_NAME)=M26-LHneg.mzXML
SUBJECT_SAMPLE_FACTORS           	-	M28	Sample source:hippocampus | Sample_type:M2	RAW_FILE_NAME(Positive_mzXML_FILE_NAME)=M28-LHPOS.mzXML; RAW_FILE_NAME(Negative_mzXML_FILE_NAME)=M28-LHneg.mzXML
SUBJECT_SAMPLE_FACTORS           	-	M29	Sample source:hippocampus | Sample_type:M2	RAW_FILE_NAME(Positive_mzXML_FILE_NAME)=M29-LHPOS.mzXML; RAW_FILE_NAME(Negative_mzXML_FILE_NAME)=M29-LHneg.mzXML
SUBJECT_SAMPLE_FACTORS           	-	M31	Sample source:hippocampus | Sample_type:M3	RAW_FILE_NAME(Positive_mzXML_FILE_NAME)=M31-LHPOS.mzXML; RAW_FILE_NAME(Negative_mzXML_FILE_NAME)=M31-LHneg.mzXML
SUBJECT_SAMPLE_FACTORS           	-	M32	Sample source:hippocampus | Sample_type:M3	RAW_FILE_NAME(Positive_mzXML_FILE_NAME)=M32-LHPOS.mzXML; RAW_FILE_NAME(Negative_mzXML_FILE_NAME)=M32-LHneg.mzXML
SUBJECT_SAMPLE_FACTORS           	-	M33	Sample source:hippocampus | Sample_type:M3	RAW_FILE_NAME(Positive_mzXML_FILE_NAME)=M33-LHPOS.mzXML; RAW_FILE_NAME(Negative_mzXML_FILE_NAME)=M33-LHneg.mzXML
SUBJECT_SAMPLE_FACTORS           	-	M34	Sample source:hippocampus | Sample_type:M3	RAW_FILE_NAME(Positive_mzXML_FILE_NAME)=M34-LHPOS.mzXML; RAW_FILE_NAME(Negative_mzXML_FILE_NAME)=M34-LHneg.mzXML
SUBJECT_SAMPLE_FACTORS           	-	M35	Sample source:hippocampus | Sample_type:M3	RAW_FILE_NAME(Positive_mzXML_FILE_NAME)=M35-LHPOS.mzXML; RAW_FILE_NAME(Negative_mzXML_FILE_NAME)=M35-LHneg.mzXML
SUBJECT_SAMPLE_FACTORS           	-	M36	Sample source:hippocampus | Sample_type:M3	RAW_FILE_NAME(Positive_mzXML_FILE_NAME)=M36-LHPOS.mzXML; RAW_FILE_NAME(Negative_mzXML_FILE_NAME)=M36-LHneg.mzXML
SUBJECT_SAMPLE_FACTORS           	-	M38	Sample source:hippocampus | Sample_type:M3	RAW_FILE_NAME(Positive_mzXML_FILE_NAME)=M38-LHPOS.mzXML; RAW_FILE_NAME(Negative_mzXML_FILE_NAME)=M38-LHneg.mzXML
SUBJECT_SAMPLE_FACTORS           	-	M39	Sample source:hippocampus | Sample_type:M3	RAW_FILE_NAME(Positive_mzXML_FILE_NAME)=M39-LHPOS.mzXML; RAW_FILE_NAME(Negative_mzXML_FILE_NAME)=M39-LHneg.mzXML
SUBJECT_SAMPLE_FACTORS           	-	QC-1	Sample source:hippocampus | Sample_type:Control	RAW_FILE_NAME(Positive_mzXML_FILE_NAME)=QC-1-LHPOS.mzXML; RAW_FILE_NAME(Negative_mzXML_FILE_NAME)=QC-1-LHneg.mzXML
SUBJECT_SAMPLE_FACTORS           	-	QC-2	Sample source:hippocampus | Sample_type:Control	RAW_FILE_NAME(Positive_mzXML_FILE_NAME)=QC-2-LHPOS.mzXML; RAW_FILE_NAME(Negative_mzXML_FILE_NAME)=QC-2-LHneg.mzXML
SUBJECT_SAMPLE_FACTORS           	-	QC-3	Sample source:hippocampus | Sample_type:Control	RAW_FILE_NAME(Positive_mzXML_FILE_NAME)=QC-3-LHPOS.mzXML; RAW_FILE_NAME(Negative_mzXML_FILE_NAME)=QC-3-LHneg.mzXML
SUBJECT_SAMPLE_FACTORS           	-	QC-4	Sample source:hippocampus | Sample_type:Control	RAW_FILE_NAME(Positive_mzXML_FILE_NAME)=QC-4-LHPOS.mzXML; RAW_FILE_NAME(Negative_mzXML_FILE_NAME)=QC-4-LHneg.mzXML
SUBJECT_SAMPLE_FACTORS           	-	QC-5	Sample source:hippocampus | Sample_type:Control	RAW_FILE_NAME(Positive_mzXML_FILE_NAME)=QC-5-LHPOS.mzXML; RAW_FILE_NAME(Negative_mzXML_FILE_NAME)=QC-5-LHneg.mzXML
SUBJECT_SAMPLE_FACTORS           	-	QC-6	Sample source:hippocampus | Sample_type:Control	RAW_FILE_NAME(Positive_mzXML_FILE_NAME)=QC-6-LHPOS.mzXML; RAW_FILE_NAME(Negative_mzXML_FILE_NAME)=QC-6-LHneg.mzXML
#COLLECTION
CO:COLLECTION_SUMMARY            	After completing behavioral and imaging experiments, the mice were decapitated
CO:COLLECTION_SUMMARY            	under isoflurane anesthesia and their brains were immediately
CO:COLLECTION_SUMMARY            	extracted.Therefore, the hippocampus was extracted from the brain tissue based
CO:COLLECTION_SUMMARY            	on its anatomical features for further analysis and the tissue was either
CO:COLLECTION_SUMMARY            	rapidly frozen in liquid nitrogen, stored at -80°C for metabolomics analysis.
CO:SAMPLE_TYPE                   	Hippocampus
#TREATMENT
TR:TREATMENT_SUMMARY             	After one week of acclimatization, 8-week-old C57BL/6J mice were randomly
TR:TREATMENT_SUMMARY             	divided into a sham surgery group (Sham) and a photothrombotic surgery group
TR:TREATMENT_SUMMARY             	(Photothrombotic surgery, PT). We utilized a PT stroke model targeting the
TR:TREATMENT_SUMMARY             	frontal cortex of mice, which has been demonstrated to be suitable for studying
TR:TREATMENT_SUMMARY             	post-stroke dementia. Mice in the PT group were anesthetized with 2% isoflurane
TR:TREATMENT_SUMMARY             	and maintained under general anesthesia throughout the procedure. Rose Bengal
TR:TREATMENT_SUMMARY             	(200 µL, 10 mg/mL in sterile saline solution, Sigma-Aldrich, USA) was
TR:TREATMENT_SUMMARY             	intraperitoneally injected and circulated for 8 minutes. Subsequently, a cold
TR:TREATMENT_SUMMARY             	light source with a diameter of 4.5 mm was used to irradiate the exposed skull
TR:TREATMENT_SUMMARY             	for 15 minutes, positioned 3.5 mm to the left of the bregma, targeting the left
TR:TREATMENT_SUMMARY             	frontal lobe. The Sham group underwent the same surgical procedure but was
TR:TREATMENT_SUMMARY             	administered 200 µL of sterile saline (0.9% NaCl, Pfizer, Australia) instead of
TR:TREATMENT_SUMMARY             	Rose Bengal. Both the Sham and PT groups were then further divided based on the
TR:TREATMENT_SUMMARY             	time post-stroke into T1 (C1 n=11, M1 n=11), T2 (C2 n=11, M2 n=11), and T3 (C3
TR:TREATMENT_SUMMARY             	n=11, M3 n=11) groups, where C1, C2, and C3 represent the Sham groups at 14, 32,
TR:TREATMENT_SUMMARY             	and 84 days post-stroke, respectively, and M1, M2, and M3 represent the PT
TR:TREATMENT_SUMMARY             	groups at 14, 32, and 84 days post-stroke, respectively.
#SAMPLEPREP
SP:SAMPLEPREP_SUMMARY            	Take 10 mg of mouse hippocampus in a 1.5 mL EP tube, add 400 µL of cold
SP:SAMPLEPREP_SUMMARY            	acetonitrile:methanol (v:v = 1:1) to 200 µL of plasma to precipitate proteins,
SP:SAMPLEPREP_SUMMARY            	mix for 60 seconds, and then centrifuge to collect the supernatant. Sequentially
SP:SAMPLEPREP_SUMMARY            	add methanol:water:dichloromethane (v:v:v = 1:1:2), three mass
SP:SAMPLEPREP_SUMMARY            	spectrometry-grade extraction solvents, homogenize at 60 Hz for 2 minutes, and
SP:SAMPLEPREP_SUMMARY            	let stand at 4°C for 30 minutes. Centrifuge at 4°C, 15000 rpm for 15 minutes.
SP:SAMPLEPREP_SUMMARY            	The liquid separates into two layers: proteins and tissue precipitate in the
SP:SAMPLEPREP_SUMMARY            	middle, the upper layer is a methanol-water solution mainly containing polar
SP:SAMPLEPREP_SUMMARY            	small molecules, and the lower layer is a dichloromethane solution mainly
SP:SAMPLEPREP_SUMMARY            	composed of lipid-soluble small molecules. Take the upper layer solution, dry it
SP:SAMPLEPREP_SUMMARY            	under N2, and reconstitute it with 200 µL of 50% acetonitrile aqueous solution
SP:SAMPLEPREP_SUMMARY            	containing an internal standard (2-chlorophenylalanine). Centrifuge at 4°C,
SP:SAMPLEPREP_SUMMARY            	15000 rpm for 15 minutes, then draw 60 µL and inject it into a liquid phase
SP:SAMPLEPREP_SUMMARY            	vial with a glass liner for analysis. Take the lower layer solution, dry it
SP:SAMPLEPREP_SUMMARY            	under N2, and reconstitute it with 100 µL of chloroform
SP:SAMPLEPREP_SUMMARY            	(dichloromethane):methanol = 1:1 solution containing an internal standard
SP:SAMPLEPREP_SUMMARY            	(2-chlorophenylalanine). Centrifuge at 4°C, 15000 rpm for 15 minutes, then draw
SP:SAMPLEPREP_SUMMARY            	100 µL and inject it into a liquid phase vial with a glass liner for analysis.
SP:SAMPLEPREP_SUMMARY            	Simultaneously, take 10 µL of all supernatant to prepare Quality Control (QC)
SP:SAMPLEPREP_SUMMARY            	samples to monitor the stability of the instrument acquisition method.
#CHROMATOGRAPHY
CH:CHROMATOGRAPHY_TYPE           	Reversed phase
CH:INSTRUMENT_NAME               	Waters Acquity
CH:COLUMN_NAME                   	Phenomenex Kinetex C18 (100 x 2.1 mm, 2.6 µm)
CH:SOLVENT_A                     	100% Water; 5 mM ammonium acetate; 0.1% formic acid
CH:SOLVENT_B                     	100% Acetonitrile
CH:FLOW_GRADIENT                 	0-0.5 min 98% B；0.5-13 min，98-40% B；13-3.1 min，40-98% B；13.1-18
CH:FLOW_GRADIENT                 	min，98-98%
CH:FLOW_RATE                     	0.3 mL/min
CH:COLUMN_TEMPERATURE            	35°C
#ANALYSIS
AN:ANALYSIS_TYPE                 	MS
#MS
MS:INSTRUMENT_NAME               	ABI Sciex 6600 TripleTOF
MS:INSTRUMENT_TYPE               	Triple TOF
MS:MS_TYPE                       	ESI
MS:ION_MODE                      	NEGATIVE
MS:MS_COMMENTS                   	The electrospray ionization (ESI) negtive ion mode was employed, with a mass
MS:MS_COMMENTS                   	detection range of 80-1000 m/z, and the collision energy for secondary fragments
MS:MS_COMMENTS                   	was set at -40±15 V. After data acquisition, the raw data were preprocessed
MS:MS_COMMENTS                   	using Markerview software. The main parameters included: peak extraction time of
MS:MS_COMMENTS                   	0.5-15 minutes, retention time deviation of 0.1 minutes, mass-to-charge ratio
MS:MS_COMMENTS                   	deviation of 10 ppm, and peak intensity >500. Metabolites with an overall
MS:MS_COMMENTS                   	detection rate of less than 80% were excluded from the analysis. An Excel
MS:MS_COMMENTS                   	spreadsheet containing m/z, retention time (RT), and peak intensity was
MS:MS_COMMENTS                   	exported. Prior to the next step of analysis, peaks with intensities below 500
MS:MS_COMMENTS                   	were excluded. The coefficient of variation (CV: standard deviation/mean ×
MS:MS_COMMENTS                   	100%) was calculated, and peaks with CV% >30% were identified as
MS:MS_COMMENTS                   	high-variability peaks and thus also excluded. Multivariate statistical analysis
MS:MS_COMMENTS                   	of the data was performed using SMICA-P software (v14.0, Umetrics, Umea,
MS:MS_COMMENTS                   	Sweden), primarily including principal component analysis (PCA). PCA is an
MS:MS_COMMENTS                   	unsupervised model used to examine changes in the metabolic patterns of the
MS:MS_COMMENTS                   	mouse hippocampus. Significant differential metabolites were screened through
MS:MS_COMMENTS                   	univariate statistical analysis, and volcano plots were generated to represent
MS:MS_COMMENTS                   	these findings. Metabolite identification was primarily based on accurate m/z
MS:MS_COMMENTS                   	values and MS/MS characteristic fragments, using software such as MS-DIAL (Mass
MS:MS_COMMENTS                   	spectrometry-data independent analysis software), MetDNA2
MS:MS_COMMENTS                   	(http://metdna.zhulab.cn/), and One-MAP (http://www.5omics.com/). These
MS:MS_COMMENTS                   	identified metabolites were further validated using HMDB (https://hmdb.ca/).
MS:MS_COMMENTS                   	Pathway enrichment analysis was conducted via Metaboanalyst
MS:MS_COMMENTS                   	(https://www.metaboanalyst.ca/).
MS:MS_RESULTS_FILE               	ST003767_AN006184_Results.txt	UNITS:Peak area	Has m/z:Yes	Has RT:Yes	RT units:Minutes
#END