Summary of Study ST002968

This data is available at the NIH Common Fund's National Metabolomics Data Repository (NMDR) website, the Metabolomics Workbench, https://www.metabolomicsworkbench.org, where it has been assigned Project ID PR001847. The data can be accessed directly via it's Project DOI: 10.21228/M8ZT64 This work is supported by NIH grant, U2C- DK119886.

See: https://www.metabolomicsworkbench.org/about/howtocite.php

This study contains a large results data set and is not available in the mwTab file. It is only available for download via FTP as data file(s) here.

Study ID	ST002968
Study Title	Untargeted metabolomic studies of the PSD95-nNOS uncoupling agent against post-stroke depression in rats
Study Type	research
Study Summary	To elucidate the antidepressant mechanisms of the PSD95-nNOS decoupler ZL006 using an innovative integrated metabolomics approach.
Institute	Nanjing University of Science & Technology
Department	Center of Molecular Metabolism
Last Name	Hu
First Name	Yudie
Address	Nanjing University of Science and Technology
Email	huyudie@njust.edu.cn
Phone	+8618759270506
Submit Date	2023-11-08
Raw Data Available	Yes
Raw Data File Type(s)	mzML
Analysis Type Detail	LC-MS
Release Date	2023-11-28
Release Version	1

Select appropriate tab below to view additional metadata details:

Project:

Project ID:	PR001847
Project DOI:	doi: 10.21228/M8ZT64
Project Title:	Untargeted metabolomic studies of the PSD95-nNOS uncoupling agent against post-stroke depression in rats
Project Type:	research
Project Summary:	To investigate the effect of nNOS-PSD95 decoupler ZL006 on PSD and its mechanism of action.
Institute:	Nanjing University of Science & Technology
Department:	Center of Molecular Metabolism
Last Name:	Hu
First Name:	Yudie
Address:	Nanjing University of Science and Technology
Email:	huyudie@njust.edu.cn
Phone:	+8618759270506

Subject:

Subject ID:	SU003081
Subject Type:	Mammal
Subject Species:	Rattus norvegicus
Taxonomy ID:	10116

Factors:

Subject type: Mammal; Subject species: Rattus norvegicus (Factor headings shown in green)

mb_sample_id	local_sample_id	Treatment
SA323006	QC3	blank
SA323007	QC2	blank
SA323008	QC4	blank
SA323009	QC1	blank
SA323010	QC5	blank
SA323011	QC6	blank
SA323012	ZH2	high
SA323013	ZH1	high
SA323014	ZH3	high
SA323015	ZH5	high
SA323016	ZH4	high
SA323017	ZH6	high
SA323018	ZD6	low
SA323019	ZD5	low
SA323020	ZD4	low
SA323021	ZD1	low
SA323022	ZD2	low
SA323023	ZD3	low
SA323024	M2	model
SA323025	M1	model
SA323026	M4	model
SA323027	M3	model
SA323028	M5	model
SA323029	M6	model
SA323030	S3	sham
SA323031	S2	sham
SA323032	S4	sham
SA323033	S6	sham
SA323034	S1	sham
SA323035	S5	sham

Showing results 1 to 30 of 30

Showing results 1 to 30 of 30

Collection:

Collection ID:	CO003074
Collection Summary:	After one week of adaptation (290-320g), the rats underwent middle cerebral artery occlusion (MCAO) surgery to induce focal cerebral ischemia: referring to literature, the thread embolism method was used to induce focal cerebral ischemia for 60 minutes. Rats were evaluated by Longa scoring at 1h and 24h after surgery. Rats with Longa scores of 1-3 were included in subsequent experiments. Penicillin was given daily to the rats for 3 consecutive days after surgery. 3 days later, chronic unpredictable mild stress (CUMS) combined with social isolation modeling method was used to establish the PSD model. The CUMS model included: day-night reversal for 36 h; water deprivation for 18 h; food deprivation for 24h; humid environment for 24 h; 45° inclined cage for 18h; forced swimming in 4°c cold water for 5 min; 45°c stimulation for 5 min; tail pinch for 1 min. Rats were subjected to one of the stimuli randomly at a fixed time every day in an unpredictable way. Each stimulus was used no more than 4 times over 3 weeks. The rats in the Sham group were group-housed with 4-6 rats per cage. Rats that died or underscored during the experiment were replaced by the same batch of rats, the success rate of MCAO surgery was 83%. After 21 days of CUMS, the low-dose and high-dose ZL006 groups received intraperitoneal injection of prepared drug solution (10% ZL006-DMSO solution, 90% ethanol: corn oil: saline = 1:1:8, 2% Tween-80), while the Sham and Model groups received the same volume of solvent.
Sample Type:	Brain cortex

Treatment:

Treatment ID:	TR003090
Treatment Summary:	The rats were divided into 4 groups: Sham operation (Sham, S) group, Model (M) group, low-dose ZL006-10mg/kg (ZD) group, high-dose ZL006-20mg/kg (ZH) group.

Sample Preparation:

Sampleprep ID:	SP003087
Sampleprep Summary:	After euthanizing the rats, the brains were rapidly removed and the brain cortical tissues were separated on ice and stored at -80°C. 30 mg of each sample was added to 450 μL pre-cooled MeOH: H2O (4:1, v/v) and zirconium beads. The homogenizer was pre-cooled and homogenized at 50 Hz for 3 × 15 s with an interval of 5 s. After standing at -20°C for 1 h, samples were centrifuged at 16000 g and 4°C for 15 min. The supernatant was collected, a portion of each brain tissue sample was taken as quality control, the remaining samples and QC were evaporated to 4/5 of the original volume under a nitrogen blower. Excess water was removed using a freeze dryer and stored at -80°C. The samples were reconstituted with 200 μL methanol/water (1:1, v/v). After vortexing for 30 s and ultrasonicating on ice for 1 min, samples were centrifuged at 16000 g and 4°C for 15 min. The supernatant was transferred to UPLC injection vials for LC-MS analysis.

Sampleprep ID:

SP003087

Sampleprep Summary:

After euthanizing the rats, the brains were rapidly removed and the brain cortical tissues were separated on ice and stored at -80°C. 30 mg of each sample was added to 450 μL pre-cooled MeOH: H2O (4:1, v/v) and zirconium beads. The homogenizer was pre-cooled and homogenized at 50 Hz for 3 × 15 s with an interval of 5 s. After standing at -20°C for 1 h, samples were centrifuged at 16000 g and 4°C for 15 min. The supernatant was collected, a portion of each brain tissue sample was taken as quality control, the remaining samples and QC were evaporated to 4/5 of the original volume under a nitrogen blower. Excess water was removed using a freeze dryer and stored at -80°C. The samples were reconstituted with 200 μL methanol/water (1:1, v/v). After vortexing for 30 s and ultrasonicating on ice for 1 min, samples were centrifuged at 16000 g and 4°C for 15 min. The supernatant was transferred to UPLC injection vials for LC-MS analysis.

Combined analysis:

Analysis ID	AN004876	AN004877
Analysis type	MS	MS
Chromatography type	Reversed phase	Reversed phase
Chromatography system	Shimadzu 20AD	Shimadzu 20AD
Column	Waters ACQUITY UPLC BEH C18 (100 x 2.1mm,1.7um)	Waters ACQUITY UPLC BEH C18 (100 x 2.1mm,1.7um)
MS Type	ESI	ESI
MS instrument type	Triple TOF	Triple TOF
MS instrument name	ABI Sciex 5600+ TripleTOF	ABI Sciex 5600+ TripleTOF
Ion Mode	POSITIVE	NEGATIVE
Units	log2(peak area)	log2 (peak area)

Chromatography:

Chromatography ID:	CH003679
Instrument Name:	Shimadzu 20AD
Column Name:	Waters ACQUITY UPLC BEH C18 (100 x 2.1mm,1.7um)
Column Temperature:	40
Flow Gradient:	0-3 min，75%B；3-6 min，75%B-70%B；6-7 min，70%B-65%B；7-10 min，65%B-40%B；10-10.1 min，40%B-95%B；10.1-15 min，95%B
Flow Rate:	0.3 mL/min
Solvent A:	100% water; 0.1% formic acid
Solvent B:	100% acetonitrile
Chromatography Type:	Reversed phase

MS:

MS ID:	MS004620
Analysis ID:	AN004876
Instrument Name:	ABI Sciex 5600+ TripleTOF
Instrument Type:	Triple TOF
MS Type:	ESI
MS Comments:	Data was collected in ESI positive ionization mode. The conditions of the ESI source were set as follows: nitrogen was used as nebulizer gas and auxiliary gas, gas 1 (GS1) 55 psi; auxiliary gas (GS2), 55 psi; curtain gas, 35 psi; ion source temperature, 550°C; spray voltage, 5500 V (+). In TOF MS-IDA-MS/MS acquisition, the TOF MS mass scanning range was 100-1250 m/z, and the accumulation time was set to 0.10 s/spectrum. The product ion scanning mass range was 50-1250 m/z, and the accumulation time was set to 0.05 s/spectrum. Independent analysis (IDA) mode was used with high sensitivity mode for product ion scanning. For better spectrum quality in IDA mode, the parameters were set as follows: declustering potential -80 V, collision energy -35 ± 15eV, ion intensity not less than 100 cps, isotopes excluded within 4 Da, ion tolerance 50 mDa, maximum 10 candidate ions monitored per cycle, dynamic background subtraction turned on to improve sensitivity for low abundance or trace analytes. External calibration was performed every 6 samples for TOF MS and TOF MS/MS automatic calibration.
Ion Mode:	POSITIVE

MS ID:	MS004621
Analysis ID:	AN004877
Instrument Name:	ABI Sciex 5600+ TripleTOF
Instrument Type:	Triple TOF
MS Type:	ESI
MS Comments:	Data was collected in ESI negative ionization mode. The conditions of the ESI source were set as follows: nitrogen was used as nebulizer gas and auxiliary gas, gas 1 (GS1) 55 psi; auxiliary gas (GS2), 55 psi; curtain gas, 35 psi; ion source temperature, 550°C; spray voltage, -4500 V (-). In TOF MS-IDA-MS/MS acquisition, the TOF MS mass scanning range was 100-1250 m/z, and the accumulation time was set to 0.10 s/spectrum. The product ion scanning mass range was 50-1250 m/z, and the accumulation time was set to 0.05 s/spectrum. Independent analysis (IDA) mode was used with high sensitivity mode for product ion scanning. For better spectrum quality in IDA mode, the parameters were set as follows: declustering potential -80 V, collision energy -35 ± 15eV, ion intensity not less than 100 cps, isotopes excluded within 4 Da, ion tolerance 50 mDa, maximum 10 candidate ions monitored per cycle, dynamic background subtraction turned on to improve sensitivity for low abundance or trace analytes. External calibration was performed every 6 samples for TOF MS and TOF MS/MS automatic calibration.
Ion Mode:	NEGATIVE