动物造模,伤性癫痫动物模型,药效评价 z-bio 2024-09-05 1031

　　1. Animal modeling materials: Healthy adult male SD rats, weighing 180-220g; Medication: 10% chloral hydrate, FeCl2 solution.

　　2. Modeling method: Rats were anesthetized intraperitoneally with 10% chloral hydrate (350mg/kg). Fixed on a stereotactic frame. Surgical exposure of the skull and determination of the drilling site based on the injection site. Drill open the skull, and according to the stereotactic map of the rat brain by Bao Xinmin et al. (1991), slowly inject the micro injector to the target point (the coordinates of the right frontal cortex of the rat are: 2.0mm in front of the anterior fontanelle, 3.0mm beside the midline, and 2.0mm below the dura mater). Drill a hole on the right forehead and right pillow to install stainless steel electrodes and secure them. Place the reference electrode in the right ear. Connected to the POWERLAB physiological recorder. Start recording the electroencephalogram, and after 10 minutes, slowly inject 1000nmol of FeCl2 (0.1mol/L) into the target using an automatic thruster at a rate of 1 μ L/min, leave the needle for 5 minutes after injection. The sham surgery control group was injected with an equal amount of physiological saline at the same site.

　　3. Modeling principle: Iron has an epileptic effect.

　　4. Changes after modeling. After successful injection of FeCl2, the seizure was characterized by paroxysmal gaze and immobility in rats, followed by wet dog like movements. Or automatic symptoms may occur. Repeated, stereotyped nodding, chewing, yawning, sneezing, accompanied by facial twitching, and mild convulsions, resulting in tremors in one forelimb: standing, tremors in both forelimbs, and persistent nodding; Increased tremors in both forelimbs. Loss of balance and falling resulting in generalized ankylosing and clonic seizures; Severe cases may experience exhaustion leading to death.

　　The disease course of the model group is clearly divided into acute phase, stationary phase, and chronic phase. Frequent seizures occur within 7 days, especially within 3 days, followed by a decrease in seizures. After 15 days, more regular seizures occur, with focal seizures being more common.

　　The EEG frequency of normal rats is mainly 5-10Hz, and the amplitude is less than 200V. With low amplitude β、α Mainly, scattered θ Wave. After injection of physiological saline, there was no significant change in the EEG waveform. After injecting the cortex into the model group, the average latency period was (60+18) seconds, and the EEG of the rats showed various forms of epileptic like discharge waveforms. Typical waveforms include single spike, multiple spike, multiphase spike, biphasic spike, normal spike, spike rhythm, slow wave, spike slow wave, slow wave superimposed spike rhythm, sharp wave, episodic rhythm wave, and post seizure suppression. The fastest frequency can reach 30Hz. The wave amplitude can reach up to 310V, and inhibitory waves may appear after the onset.

　　5. Pathological changes after modeling: The general appearance of the model group showed right frontal lobe atrophy. Obvious brownish yellow color, indicating deposition of hemosiderin, while no significant changes were observed in the corresponding parts of the control group. Observation under light microscope: The injection site of the model group showed significant neuronal degeneration and death, as well as cellular structure disintegration and disorder. The experimental side showed significant loss of hippocampal neurons, with the CA3 area being the most prominent. Neurons in the CA3 and CA4 areas of the experimental side of the hippocampus were missing, with nuclei pyknosis and staining, and eosinophilic neurons were present with disordered cell arrangement; Glial cell proliferation. Mild on the control side; CA1 and CA2 lesions are mild. The model group shows cerebral cortex and hippocampus CA1 and CA3