小鼠耐药蛋白的测定及耐药癫（癎）所致脑损伤的研究

目的 制作小鼠耐药癫（癎）模型,考察耐药蛋白的表达以及实验动物脑损伤情况,为进一步耐药癫（癎）逆转研究奠定基础.方法 取10只小鼠,给予戊四氮制作小鼠癫（癎）模型,造模成功后,低剂量持续口服灌胃给苯妥英钠制作耐药癫（癎）模型,试验期间考察小鼠癫（癎）的发作级别和频率,以确定小鼠耐药癫（癎）模型制作是否成功.另取20只小鼠进行后续研究,将其随机分为2组,分别为阴性对照组及耐药癫（癎）模型组.阴性对照组给予生理盐水,耐药癫（癎）模型组按前述方法造模.结束后将全部小鼠处死取脑,采用Western blot法考察小鼠脑内P-糖蛋白的变化,采用HE染色法考察耐药癫（癎）小鼠脑损伤情况,采用免疫组化法考察脑内Caspase-3的表达情况.结果 耐药癫（癎）模型组中10只小鼠有8只造模成功,进入试验.脑组织Western blot试验表明,耐药癫（癎）模型组较阴性对照组小鼠脑内P-糖蛋白的表达显著增高（P〈0.01）;脑组织HE染色显示,耐药癫（癎）模型组呈现神经细胞核固缩、变形,神经细胞呈现部分水肿,并有神经细胞变性、死亡,与对照组相比差异有统计学意义（P〈0.05）;免疫组化结果显示,耐药癫（癎）模型组呈现Caspase-3的表达（P〈0.05）.结论 耐药癫（癎）小鼠呈现脑内P-糖蛋白高表达,并出现脑组织损伤.

Determination on the resistance protein and brain damage in drug-resistant epilepsy mice

Objective To produce resistant epilepsy model in mice, lay the foundation for further drug-resistant epilepsy reversion. Methods 10 mice were given pentylenetetrazol, and then low-dose continuous oral phenytoin sodium was given to produce a mouse drug-resistant epilepsy model, the attack level of epilepsy arid seizure frequency in mice was inspected to determine the success of drug-resistant epilepsy model making. Another 20 mice were randomly divided into 2 groups：negative control group received saline, drug-resistant epilepsy model group followed by the aforementioned method. At the end of the modeling, the mice were sacrificed to take the brain and study the changes of P-glycoprotein by Western blot method, HE staining was used to examine the brain injury in mice. Immunohistochemical methods were used to investigate brain caspase-3 expression. Results 80% of the mice met the inclusion criteria of drug-resistant epilepsy after the modeling. In subsequent studies, 8 mice of 10 mice model successful entered the experiment in drug-resistant epilepsy model group. The Western blot experiments of brain tissue showed that P-glycoprotein expression was significantly higher in drug-resistant epilepsy model group than that of the negative control group（ P 〈 0. 01 ）. Brain tissue HE staining showed that drug-resistant epilepsy model group had nerve cell nucleus shrinkage, deformation, edema of nerve cells, nerve cell degeneration and death, there were significant differences compared with control group（ P 〈0. 05 ）. Immunohistochemistry results showed that caspase-3 expression existed in drug-resistant epilepsy model group（ P 〈 0. 05 ）. Conclusion The method is able to make mice resistant epilepsy model successfully at a high success rate, and drug-resistant epilepsy mice show high expression of brain P-glycoprotein, and brain tissue damage.