大鼠模型,动物模型 z-bo 2020-07-29 592

　　Objective: To use lithium chloride-pilocarpine to prepare a Wistar rat epilepsy model, and to study the appropriate dosage and usage of pilocarpine. Methods: Wistar rats were injected intraperitoneally with 3 mmol/kg lithium chloride, and 24 hours later, different doses of pilocarpine were injected intraperitoneally. Results: The incidence and mortality of persistent epilepsy (SE) in the 40 mg/kg and 30 mg/kg single injection groups were 100%, while the success rate of the chronic relapsing epilepsy (SRS) model was 0%. At mg/kg, the incidence of SE in the three injection groups was 28%, the mortality rate was 10%, and the success rate of SRS was 18%; the incidence of SE in the 25 mg/kg 3 injection group was 10%. , The mortality rate is 0%, and the SRS success rate is 10%. Conclusion: Lithium chloride-pilocarpine can be used to prepare Wistar rat epilepsy model. The dose of lucarpine should be 25-30 mg/kg. The split injection method is better than the single injection method.

　　Epilepsy is a common disease. Epilepsy is unpredictable, and the degree of seizures cannot be controlled artificially, making research difficult. Therefore, it is very important to establish experimental animal epilepsy models, study the mechanism of epilepsy, and screen and evaluate the effects of antiepileptic drugs.

　　There are many ways to prepare animal epilepsy models. It is reported in the literature that the rat status epilepticus (SE) caused by lithium chloride-pilocarpine injection reaches 1 hour, and the survivors have a spontaneous epilepsy (SRS) model [1, 2], which is similar to the most common adult epilepsy The type of epilepsy, temporal lobe epilepsy, has the advantages of ease of use, high repeatability and low external interference. In this experiment, we used lithium chloride-pilocarpine to create a Wistar rat epilepsy model, and described the specific dosage and usage of pilocarpine. 1 Materials and methods 1.1 Experimental animals and a group of healthy female Wistar rats (weight 210-260 grams) were purchased from the Experimental Animal Breeding Center of the Chinese Academy of Medical Sciences, and provided adequate food and water at room temperature and natural light. It is 40 mg/kg single injection group (6), 40 mg/kg single injection group (4), 30 mg/kg single injection group (5), 30 mg/kg depends on the injection dose of pilocarpine injection group Divided into 3 times (50 animals), 25mg/kg 3 times injection group (10 animals). 1.2 Establishment of epilepsy model 24 hours before the experiment, all rats were injected intraperitoneally with 3 mmol/kg lithium chloride (concentration 1.5 mol/L) [3]. Twenty-four hours later, different doses of pilocarpine (at a concentration of 0.1%) were injected intraperitoneally. In the separate injection group, the interval between each injection is 10 minutes [4]. Patients with persistent seizures (SE) lasting 1 hour will be discontinued by intraperitoneal injection of disi epa 10 mg/kg [2].

　　2 results

　　2.1 Different performance statistics for each dose group All rats developed cholinergic peripheral nerve irritation symptoms 5-10 minutes after the first injection of pilocarpine: erect hair, salivation, hematuria. And manifested in varying degrees, including: lack of movement, gaze, automatic mouth, nodding, blinking and dog-like dampness; 33 rats transient epileptic seizures: transient seizures manifested as erections, fibrous plexus in the forelimbs and head, Decrease 30 minutes after the first injection, each lasting 30 to 45 seconds, then every 2 to 5 minutes. I have another episode. 30 rats developed persistent epilepsy (1 hour). The seizures were the same; 20 rats died 1-2 days after the persistent grand mal seizure; all 10 survivors after the SE seizure had spontaneous seizures within 20-60 days. However, every swing, bite and stare movements last from a few seconds to tens of seconds.

　　Statistical analysis of the SRS success rate of 25mg/kg3 injection group and 30mg/kg3 injection group (statistical method is χ2 test, χ2=0.38): P\→0.05, that is, there is no difference in statistical difference between the two methods.

　　"The above results show that the Wistar rat epilepsy model is made of lithium chloride-pilocarpine. The total amount of pilocarpine is 25-30 mg/kg. A dose of 40 mg/kg can cause SE, but it will cause death. Very high; if the total dose is 30 mg/kg, the mortality rate of a single injection is higher, so it should be injected in divided doses. It can be scaled proportionally according to different mouse performance to adapt to different individual differences.

　　3 Discussion

　　The Wistar rat epilepsy model established by intraperitoneal injection of lithium chloride-pilocarpine has five behavioral levels described by Racine [5]. That is, mouth and face cramps; II head-to-head movement; III nose nu of the forelimbs; IV clonic erection; V loss of postural control (falls). Among them, I-III behaviors represent complex partial seizures, and IV-V behaviors represent secondary generalized seizures. This model usually simulates the entire process of human epilepsy, spread and formation. This is the only animal model that matches the most common adult epilepsy, temporal lobe epilepsy. [6]: Easy to operate and reproducible, it is an ideal model for studying the development and formation of epilepsy. According to reports, using

　　When preparing the Wistar rat epilepsy model with lithium chloride-pilucapine, a single injection of 30 mg/kg pilocarpine can cause SE in 70% of the rats, but SE lasts for 90 minutes, but ends with barium. The mortality rate is still 45%. 30 mg/kg pilocarpine was injected in batches, and the mortality after the attack dropped to less than 10%. After repeated low-dose injections, the incidence of SRS is higher than a single high-dose injection [7], so the small-dose divided injection method is better than a large-dose single injection. The results of this experiment are consistent with those reported in the literature. Compared with reports in the literature, the epilepsy model in this experiment has a lower success rate and a higher mortality rate. The possible reason is that the experiment is all female rats, and most of the rats in the literature are male rats [1,2,6], and the sex is different. Rats have different sensitivities to pilocarpine; according to reports in the literature, agonistic neurons begin to work in the second week of life in rats, and acetyltransferase reaches 83% of rats within four weeks. The sensitivity of calpain to epilepsy is related to age [8]. When the mice are three weeks old, the mortality rate is highest, and the mortality rate of adult rats decreases. The adult rats used in this experiment are adult rats, but they are better than those reported in the literature. Slightly smaller (the weight of the above-mentioned experimental mice is 210-260g, most of the literature is 250-300g), this may be one of the reasons. Atropine may antagonize the epilepsy caused by pilocarpine and the damage to brain tissue. Atropine was not given in this experiment.