Return to study ST002968 main page
MB Sample ID: SA323030
Local Sample ID: | S3 |
Subject ID: | SU003081 |
Subject Type: | Mammal |
Subject Species: | Rattus norvegicus |
Taxonomy ID: | 10116 |
Select appropriate tab below to view additional metadata details:
Subject:
Subject ID: | SU003081 |
Subject Type: | Mammal |
Subject Species: | Rattus norvegicus |
Taxonomy ID: | 10116 |
Factors:
Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
---|---|---|---|---|
S3 | SA323030 | FL038532 | sham | Treatment |
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. |
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 |