Rat Model for Irritable Bowel Syndrome (IBS)
- Product No.DSI533Ra01
- Organism SpeciesRattus norvegicus (Rat) Same name, Different species.
- Prototype SpeciesHuman
- SourceInduced by wrap restraint stress
- Model Animal StrainsSD Rats(SPF), healthy, male, body weight 180g~200g.
- Modeling GroupingRandomly divided into six group: Control group, Model group, Positive drug group and Test drug group.
- Modeling Period4-6 weeks
- Modeling MethodMethod:IBS-D model was established by restraint stress.
Rats were adaptable for a week and 12h fasted before the experiment.
Day1:The silicone tube (8cm from the anus) was inserted into the anus and 40 mg/L acetic acid was injected into the colon. Then the silicone tube was slowly pulled out. The anus was pressed by hand and the tail of the rats was raised 30S. Then the colon was washed with 0.01 mol/L PBS 1mL.
Day7-Day9. Rats were given restraint stress. Rats were placed in a restraint cage for 30 minutes before the experiment (in a special transparent cylindrical cylinder that restrained their limbs but did not affect their respiration). Control Group was fed in a quiet, warmand glare free environment, feeding and drinking freely. - ApplicationsCan be used to develop medicine for the prevention and treatment of IBS, such as probiotics and so on.
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Model Evaluation of the Rat Model for Irritable Bowel Syndrome (IBS)
1. General observation:
The fur color, mental state and activity of rats were observed.
2. Calculation of body mass growth rate:
After the adaptive feeding, the body weight of the experimental rats was taken as the initial body weight. The daily body weight was observed during the experiment, and the growth rate of body weight was obtained.
Body mass growth rate = (body mass per body mass) / initial mass x100%.
3. Fecal particles
The stool samples were collected from Day1, Day3, Day5, and Day7-9 for 2 hours (fixed time point 9:00-11:00).
4. dry wet weight ratio of stool (fecal water content)
1-2 grains of feces were taken from each rat in Day1, Day3, Day5 and Day7-9. The rats were weighed wet and then baked in a 50-degree thermostat, and then weighed dry.
Fecal water content (wet weight dry weight) / wet weight x100%.
5. Abdominal wall retreat reflex (AWR score)
After the model was built, the rats were forbidden to feed water at 24 h before the experiment. After mild anesthesia in rats, the balloon catheter was inserted slowly from the anus, and the 8F catheter (catheter diameter 2.7 mm and maximum balloon volume 3 ml) were inserted from the anus.
The distal end of the balloon is about 1 cm from the anus, and is fixed at the end of the tail of the rat at the outside of the anus at 1cm. Each rat was dilated 3 times with the volume of 1, 1.5, 2 ml, and each expansion lasted 3-5 min, and the interval was 30 s (against intestinal ischemia). Each capacity is repeated 3 times, taking the average.
AWR scoring criteria:
0 points, the rats' mood was basically stable when the colorectal distention was given.
1 points, when given stimulation become unstable, occasionally twisting the head;
2 points, a slight muscle contraction of the abdomen and back abdomen but not lift off the ground;
3 points, abdomen and back muscles and abdominal contraction lifted off the ground;
4 points, the abdominal muscles contracted strongly, the abdomen was arcuate, and the abdomen and perineum were lifted from the ground.
5.The open field experiment
The box is a high field 30~40cm with a bottom length of 100 cm, and the color of the circumferential wall is black. The open field bottom is divided into 25 4 x 4 cm small squares. The camera is on the top, and the vision covers the whole field. The rats were placed in the middle of the Bay, at the same time and time the camera (5 mins). The computer tracer analysis system is used to analyze the activity state of rats in a certain period of time.
Observation index: through the number of squares (animal limbs from one into another through a lattice), the number of vertical (double forelimbs simultaneously from the animal, or double forelegs on the wall as a number of central upright), time, through the central lattice.
6. Gastric emptying test(The gastric residual rate)
Each rat was given a mixture of 2 ml Arabic gum and activated carbon mixed powder by gastric lavage at Day10. After 30 minutes, the rats were sacrificed and abdomen was opened. The stomach was taken from the door to the intestinal anastomosis. The total weight of stomach and the net weight of stomach were weighed.
Gastric emptying rate (100%): (1- (stomach total weight-stomach net weight) / carbon powder mixed paste weight) x100%.
7. The intestinal propulsion
The distances from pylorus to ileocecum (total length of small intestine) and from pylorus to the front of carbon dust (advance length of carbon dust) were measured and the propulsive percentage of small intestine was calculated.
Small intestine propulsion ratio (100%) = activated carbon propulsion distance / small intestine full length x100%.
Pathological Results of the Rat Model for Irritable Bowel Syndrome (IBS)
Collecting Colon tissue
Rats were sacrificed, colon was taken out, colon tissue was taken out 2 cm away from the hepatic entrance, divided into two parts, and fixed with a 10% neutral formaldehyde solution. The other one was stored at -80 degrees, which could be used for subsequent molecular biological tests.
Collecting serum
The rats were sacrificed and fixed on the operation table. The abdominal aorta was fully exposed. The blood was collected from the abdominal aorta by vacuum blood collection. The blood was kept at room temperature for 2 hours. Then the serum was centrifuged at 4 ~3000r for 10 minutes and stored in - 80 refrigerator.
HE stainin: colon was fixed by formalin,paraffin embedded, sectioning (5um) and HE staining for colon.
Cytokines Level of the Rat Model for Irritable Bowel Syndrome (IBS)
Statistical Analysis of the Rat Model for Irritable Bowel Syndrome (IBS)
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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Immunohistochemistry (IHC) Experiment Service
Small Animal In Vivo Imaging Experiment Service
Small Animal Micro CT Imaging Experiment Service
Small Animal MRI Imaging Experiment Service
Small Animal Ultrasound Imaging Experiment Service
Transmission Electron Microscopy (TEM) Experiment Service
Scanning Electron Microscope (SEM) Experiment Service
Learning and Memory Behavioral Experiment Service
Anxiety and Depression Behavioral Experiment Service
Drug Addiction Behavioral Experiment Service
Pain Behavioral Experiment Service
Neuropsychiatric Disorder Behavioral Experiment Service
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