动物造模,药效评价,外伤后癫痫 z-bio 2024-08-06 277

　　1. Modeling material animal: Wistar rats, male, weighing 200-250g; Medication: pentobarbital, FeCl3 solution; Instruments: rat stereotaxic apparatus, wireless Bluetooth EEG monitoring system, high-speed dental drill, tooth support powder, 1 PVC thin tube with an inner diameter of 2mm (used as an introduction tube), and several acupuncture and moxibustion needles.

　　2. Modeling method: After weighing the rats, they were anesthetized by intraperitoneal injection of 2% pentobarbital solution (50mg/kg) and fixed on a stereotaxic device. Cut off the hair on the top of the head, disinfect the skin on the top of the head, and then cut open the scalp along the midline of the head, with a length of about 3cm. Peel off the fascia on the top of the head and expose the skull. Drill a 4mm diameter hole with a high-speed dental drill 3mm behind the coronal suture and 2mm beside the sagittal suture on the left skull, and make a star shaped incision of the dura mater. Additionally, drill 5 more holes for placing electrodes. Insert cotton into the tip of the introduction tube (to prevent the FeCl3 solution from overflowing too quickly) and lightly contact it with the pia mater. Inject a FeCl3 solution with a concentration of 100mmol/L and a pH of 1.5 into the tube. Connect the positive and negative electrodes of a power supply with an output current of 100 μ A to a acupuncture and moxibustion needle respectively. The acupuncture and moxibustion needle connected to the positive pole of the power supply was inserted into the FeCl3 solution in the tubule, and the acupuncture and moxibustion needle connected to the negative pole of the power supply was inserted under the skin of the right forelimb of the rat. Power on for 10 minutes.

　　EEG recording: Place a blunt and round silver wire at the front end outside the dura mater of the drilled electrode placement hole, firmly fix the silver wire to the skull with dental powder, and connect the other end of the silver wire to a wireless Bluetooth EEG monitoring system to monitor the rat's EEG.

　　3. The principle of modeling is that iron has an epileptic effect.

　　4. Changes after modeling: During anesthesia (within 1.5 hours after surgery), rats occasionally experience slight body tremors. Within about 30 minutes after waking up, the rats showed less activity, occasionally exhibiting "wet dog like shaking" and scratching their heads. During the period of 0.5-1 hour after awakening, rats begin to exhibit automatic chewing. Manifesting as rapid jaw biting and alternating facial muscle twitching, accompanied by a rhythm consistent with facial muscle twitching, the eyes of both eyes undergo large fission and protrude outward for about 1-2 minutes, occurring approximately every 20-30 minutes. After the model is established, it occurs approximately every 5 to 10 minutes between 2.5 to 4 hours. Between 5-7 hours after the establishment of the model, the frequency of epileptic seizures in rats gradually decreased, approximately once every 0.5-1 hour. After the model is established, there will be approximately one episode every 2-5 hours between 8-24 hours, mainly manifested as automatic chewing. In addition to automatic chewing, seizures in rats occasionally include leftward deviation, falls, and limb twitching. After 24 hours of model establishment, no epileptic seizures were observed.

　　At 1.5 hours after the model was established, epileptic waves appeared for the first time on the electroencephalogram. The epileptic wave is manifested as a sharp wave with significantly increased frequency and amplitude higher than the background wave. It originates from the sensory motor cortex area on the injection side, and is more pronounced in the sensory motor cortex areas on the injection side and the injection contralateral side. At the same time as the appearance of epileptic waves, rats exhibit automatic chewing behavior.

　　5. Precautions: When selecting rats for experiments, try to keep their body weight within the required range. These rats have strong tolerance and their heads are easily fixed by brain stereotaxic devices; Surgical instruments should be strictly disinfected to prevent surgical infections, surgical trauma should be minimized, and aseptic procedures should be strictly followed; When drilling a skull, attention should be paid to the depth. Once there is a sense of breakthrough, stop immediately to prevent excessive bleeding.

　　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.