Rat Model for Chronic Obstructive Pulmonary Disease (COPD)
- Product No.UDSI557Ra02
- Organism SpeciesRattus norvegicus (Rat) Same name, Different species.
- Prototype SpeciesHuman
- SourceInduced by smoking
- Model Animal StrainsSD Rats(SPF), healthy, male, body weight 190g~230g.
- Modeling GroupingRandomly divided into six group: Control group, Model group, Positive drug group and Test drug group.
- Modeling Period4 weeks
- Modeling Method1. Narcotize rats with ether at day 1 and day 14, expose its larynx, replace the tracheal tube with a venous cannula, inject 200μg/kg lipopolysaccharide (LPS) into rats trachea, then revolve rats vertically 10-20s to diffuse LPS at lung evenly.
2.At days 2-13 and days 15-28, fresh smoke was steadily provided to the closed glass box (70cm×60cm×60cm) by 15 cigarettes per day, to maintain smokescope about 100-120 mg/m3. After smoking, stimulate immediately with 5℃ cool air for 1 hour. End of modeling at day 28. - ApplicationsDiesase model
- Download n/a
- UOM Each case
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FOB
US$ 120
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Model Evaluation of the Rat Model for Chronic Obstructive Pulmonary Disease (COPD)
1.Observe activity, hair, appetite, breathe, weight of rats in each group. Weigh at 1 hour before feeding every week to observe the change of weight.
2.RT-qPCR Analysis
Collect rats PBMC and lung tissue, extract RNA, detect the level of Foxp3、RORγT by RT-PCR.
3.Western blot Analysis
Collect rats PBMC and lung tissue, extract protein, detect the level of TAZ by Western blot.
4. Flow cytometry for Peripheral blood T lymphocytes
Collect 2ml rat abdominal aortic blood in each group, extract peripheral blood mononuclear cells (PBMC) cell suspensions from rat peripheral blood lymphocytes.
(1)Detection of Th17: Add phorbol ester (50ng/ml), ionomycin (1μg/ml) and coban (2μmol/L) into the cell suspension. Incubate at 37℃, 5%CO2 for 6 hours, resuspend with 100μL PBS after washing 3 times. Add 10μL CD4 monoclonal antibody, mix it up, incubate at 4℃ without light for 30min. Wash, fix and break. Add 10μL IL-17 monoclonal antibody, incubate at 4℃ without light for 20min. Resuspend with 500μL PBS after washing repeatedly. Proportion of CD4+IL-17+ cells in CD4+T cells were detected by flow cytometry.
Detection of Treg cells: Add 10μl CD4 and CD25 monoclonal antibody into the cell suspension. After mixing, incubate at 4℃ without light for 30min. Wash, fix and break. Add 10μL FoxP3 monoclonal antibody, incubate at 4℃ without light for 20min. Resuspend with 500μL PBS after washed repeatedly. Proportion of CD4+CD25+FoxP3+ cells in CD4+T cells were detected by flow cytometry.
Pathological Results of the Rat Model for Chronic Obstructive Pulmonary Disease (COPD)
HE staining: The right lung lobe tissue was fixed on the embedded section, and the pathological changes of lung tissue were observed after staining.
Cytokines Level of the Rat Model for Chronic Obstructive Pulmonary Disease (COPD)
ELISA Analysis
Detect the concentration of IL-17 and IL-10 in serum and bronchoalveolar lavage fluid (BALF).
BALF: After blood collection, open its chest, separate mediastina, inject 3ml saline from trachea to the lung. Every time lavage thrice, collect the douche. Centrifuged it at 4℃, 3000r/min for 15 min, take the supernatant.
Serum: Place whole blood samples collected in the serum separation tube at room temperature for 2 hours or 4℃ overnight. Centrifuge it at 1,000×g for 20 min, take the supernatant. Store the supernatant at -20℃ or -80℃, avoid repeated freezing and thawing.
Statistical Analysis of the Rat Model for Chronic Obstructive Pulmonary Disease (COPD)
SPSS software is used for statistical analysis, measurement data to mean ± standard deviation (x ±s), using t test and single factor analysis of variance for group comparison, P<0.05 indicates there was a significant difference, P<0.01 indicates there are very significant differences.
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Total Anti-Oxidative Capability Assay Kit(A015-2)
Total Anti-Oxidative Capability Assay Kit (A015-1)
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UDSI557Ra01 | Rat Model for Chronic Obstructive Pulmonary Disease (COPD) | n/a |
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