抑郁症动物模型,动物造模 z-bio 2021-12-30 531

　　(1) Neurobiochemical function change model

　　method one

　　1. Modeling materials Animals: healthy purebred mice, male, 4 to 5 weeks old; drug: reserpine.

　　2. Modeling method Observe for 10 hours after injecting 2mg/kg reserpine into the tail vein of mice.

　　3. Principles of Modeling The antagonistic effect of reserpine is used to establish an animal model of depression.

　　4. General changes after modeling. Drooping eyelids occurred half an hour after the injection of reserpine, and after 1 hour, they could not move beyond the range of 5cm in diameter. The body was curled up and slow to respond to external stimuli.

　　5. Biochemical changes after modeling. The content of norepinephrine (NE) and 5-HT in the brains of mice in the control group were (220.90±3.40)ng/(ml·g), (390.99±6.93)ng/( ml·g); the contents of NE and 5-HT in the brains of the model mice were (176.03±2.70)ng/(ml·g) and (174.98±9.32)ng/(ml·g), which existed between the two groups Significant difference.

　　Method Two

　　1. Modeling materials Animals: healthy purebred mice, males, 4 to 5 weeks old; medicine: reserpine; equipment: sink.

　　2. Modeling method Inject a half dose of reserpine (1mg/kg) into the tail vein, then bind the mice’s limbs to a wooden board, immerse them in a sink at about 20℃, and deepen the water to the xiphoid process. After immersing them for 10 hours, place them Take it out for observation.

　　3. Principles of Model Making Use of environmental stimuli to cause stress and add half a dose of reserpine to create an animal model of depression.

　　4. General changes after modeling. Impatient, anxious, irritable, and anger appear in the early stage of modeling. With the continuation of the modeling time, the mice showed fatigue, slow response to external stimuli, and reduced their range of activities, gradually reaching a diameter of 5 cm.

　　5. Biochemical changes after modeling. The NE and 5-HT contents in the brains of the control group mice were (220.90±3.40)ng/(ml·g) and (390.99±6.93)ng/(ml·g) respectively; The contents of NE and 5-HT in the brain of mice were (179.89±2.70)ng/(ml·g) and (176.96±9.25)ng/(ml·g), respectively. There were significant differences between the two groups.

　　(2) Animal model of post-stroke depression

　　1. Modeling materials Animals: male Wistar rats, weighing 250-300g; drugs: anesthetics, physiological saline; equipment: hemostatic forceps, oscillators.

　　2. Modeling method Under 10% chloral hydrate (400mg/kg) anesthesia, cut open the skin of the rat's head, expose the skull, use a dental drill to drill 6 holes in the left skull, and open the left side with ophthalmic scissors For the skull, expose the dura mater on the left side of the brain. Use iris scissors to gently cut the dura mater to expose the pia mater. Use a cotton swab dipped in saline to wipe gently (under the operating microscope) until no blood vessels are visible under the microscope. skin.

　　The animals were irritated with tail clip for 1 min, water deprivation for 24 h, fasting for 24 h, 45 ℃ environment for 5 min, day and night inversion for 24 h, horizontal shaking for 30 min (160 times/min), 4 ℃ ice water swimming for 5 min and other methods. These 7 kinds of stimuli are randomly taken one every day for a total of 3 weeks. It includes: 3 times of 24h fasting, 3 times of 24h water-free, 3 times of day and night inversion, 3 times of swimming in 4℃ ice water for 5min, 3 times of 45℃ oven heating for 5min, 3 times of tail clamping for 1min, 3 times of 30min high-speed horizontal oscillation (160 times/min). The operation method of each stimulus is as follows:

　　(1) Fasting: Feeding is cut off for 24 hours, that is, no feeding will be given from 7:00 to 7:00 the next morning.

　　(2) Water prohibition: 24h water is cut off, that is, no water is fed from 7:00 am to 7:00 the next morning.

　　(3) Day and night upside-down: Put the animal in the day-night upside-down box at 7:00 in the morning, leaving the animal in a dark state without turning on the light;

　　Turn on the fluorescent lamp of the day and night inverted box until 19:00, so that the animal is in the light state, until 7:00 the next morning, take it out.

　　(4) Swimming in ice water: Put the animal in a bucket of ice water at 4℃ (water depth 15cm), the rat’s hind toes can just touch

　　and the bottom of the bucket, take out the animal 5 minutes later and put it back in the cage.

　　(5) Heat drying: the temperature of the oven is adjusted to 45°C constant, the animal is put into the oven, and the animal is taken out after 5 minutes.

　　(6) Tail clamp: Put the rat in a fixed cage, expose the tail, clamp 1cm away from the root of the tail with hemostatic forceps (do not use too much force to make the rat wailing) for 1 min, then pull the rat Put it back into the cage.

　　(7) Oscillation: Put the rat into a shaker, shake it horizontally at high speed (160 times/min) for 30 minutes and then take it out.

　　Keep animals in single cages.

　　3. Principles of Modeling The left cortex of the rat brain is removed to establish a cerebral ischemia model. On this basis, orphan and chronic unpredictable mild stress methods are used to create an animal model of post-stroke depression.

　　4. Changes after modeling. Mild dysfunction of the contralateral limb appeared immediately after waking up from anesthesia; irritability occurred 1 to 2 weeks after operation; limb function recovered in 1 week after operation; 3 weeks after operation, it was chronic After 2 weeks of stimulation, there was reduced activity, often curled up and was less active, and anhedonia, suggesting that a rat model of post stroke depression (PSD) has been established.

　　5. Precautions Strictly sterilize surgical instruments to prevent surgical infection, surgical trauma should be as small as possible, and strictly aseptic operation. Keep the animal's optimum temperature and humidity as much as possible in the breeding room.

　　(3) Rat chronic stress depression model

　　1. Modeling materials Animals: male Wistar rats, weighing 250-300g; equipment: hemostatic forceps, oscillators.

　　2. Modeling method The animals were irritated with tail clip for 1 min, water-free for 24 h, fasting for 24 h, 45 ℃ environment for 5 min, day and night inversion for 24 h, horizontal shaking for 30 min (160 times/min), 4 ℃ ice water swimming for 5 min and other methods. These 7 kinds of stimuli are randomly taken one every day for a total of 3 weeks. It includes: 3 times of 24h fasting, 3 times of 24h water-free, 3 times of day and night inversion, 3 times of swimming in 4℃ ice water for 5min, 3 times of 45℃ oven heating for 5min, 3 times of 1min tail clamping, 3 times of 30min high-speed horizontal oscillation (160 times/min). The operation method of each stimulus is as follows:

　　(1) Fasting: Feeding is cut off for 24 hours, that is, no feeding will be given from 7:00 to 7:00 the next morning.

　　(2) Water prohibition: 24h water is cut off, that is, no water is fed from 7:00 am to 7:00 the next morning.

　　(3) Day and night upside-down: Put the animal in the day-night upside-down box at 7:00 in the morning, leaving the animal in a dark state without turning on the light;

　　Turn on the fluorescent lamp of the day and night inverted box until 19:00, so that the animal is in the light state, until 7:00 the next morning, take it out.

　　(4) Swimming in ice water: Put the animal in a bucket (water depth of 15cm) filled with ice water at 4°C. The rat's hind toes can just touch the bottom of the bucket. After 5 minutes, the animal is taken out and placed in the cage.

　　(5) Heat drying: the temperature of the oven is adjusted to 45°C constant, the animal is put into the oven, and the animal is taken out after 5 minutes.

　　(6) Tail clamp: Put the rat in a fixed cage, expose the tail, clamp 1cm away from the root of the tail with hemostatic forceps (do not use too much force to make the rat wailing) for 1 min, then pull the rat Put it back into the cage.

　　(7) Oscillation: Put the rat into a shaker, shake it horizontally at high speed (160 times/min) for 30 minutes and take it out.

　　Keep animals in single cages.

　　3. Principles of Modeling Adopting orphaning methods and chronic unpredictable mild stress methods create a chronic stress depression model in rats.

　　4. After modeling, the rats with chronic stress and orphan models mainly simulated the core symptoms of human depression, that is, lack of anhedonia, which is manifested by the characteristics of reduced exercise activity, lack of pleasure, decreased exploratory behavior, and easy despair. It has a high degree of effectiveness and can last for several months. It basically meets the requirements of the depression model. It is a model widely used in foreign literature.