动物造模,药效评价,溃疡性结肠炎 z-bio 2024-08-06 296

　　1. Modeling material animal: Wistar rats, female, weighing (180 ± 10) g; Medications: pentobarbital, penicillin; Equipment: body locator, electronic stimulator.

　　2. Modeling method: After intraperitoneal injection of pentobarbital 40mg/kg anesthesia, the rats were fixed on the Jiangwan Type I stereotaxic instrument. The left and right amygdala of the rats were determined according to the Ktinig diagram, with coordinates of 30mm behind the anterior fontanelle, 48mm beside the opening, and 88mm below the dura mater. Two insulated bipolar electrodes with a diameter of 0.25mm were inserted, with the tips separated by about 0.25mm, and a micro plug was welded to the other end; The electrodes and micro plugs are fixed on the surface of the skull with dental support powder. Three consecutive days after surgery, 100000 units of penicillin were injected intraperitoneally to prevent infection.

　　Two weeks after surgery, the right amygdala was stimulated with a constant current once a day, with parameters of 400 μ A single-phase square wave, wave width of 1ms, frequency of 60Hz, and duration of 1s. Record changes in electroencephalography, as well as latency of discharge, duration of post discharge, and intensity of seizures. The latency period of discharge refers to the time from the end of stimulation to the appearance of high amplitude spike discharges in the stimulated nucleus; The post discharge duration refers to the duration of discharge in which the amplitude of the amygdala EEG wave after stimulation is at least twice that of before stimulation and the frequency is above 1Hz; The intensity of attacks is classified into behavioral stages according to the Racine method: I. immobility, eye closure, beard movement, and smell; II. Nodding and chewing accompanied by facial twitching; III. Lift one forelimb with spasms; IV. Standing with spasms in both forelimbs; V. Standing, falling, accompanied by generalized spasms. Five consecutive occurrences of the fifth phase reaction indicate complete ignition.

　　3. Modeling principle: Electrical stimulation of the amygdala can cause epilepsy in animals.

　　4. Changes after modeling: High amplitude and high-frequency epileptic discharges were observed in both the left and right amygdala after stimulation. As the number of stimuli increased, the duration of post discharge gradually prolonged, from (10 ± 2.1) s at the beginning of stimulation to (45.3 ± 5.4) s at the first appearance of phase V response; The number of stimuli required to fully ignite is (13.3 ± 4.2) times.

　　5. Precautions: Surgical instruments should be strictly disinfected to prevent surgical infections, surgical trauma should be minimized, and aseptic operation should be strictly enforced.

　　After surgery, rats should be separated and housed in cages 3-5 hours after awakening to prevent the first waking rats from licking and biting the wounds of comatose rats. Try to maintain the optimal temperature and humidity for rats in the breeding room as much as possible.