DYRK1A表达与难治性颞叶癫痫的相关性研究

The Correlation Research of DYRK1A Expression and Refractory Temporal Lobe Epilepsy

目的:通过氯化锂-匹罗卡品小剂量反复腹腔注射诱导昆明小鼠癫痫持续状态发作,模拟人类颞叶癫痫动物模型,分析DYRK1A mRNA在小鼠脑组织中的表达情况及其与海马硬化形成的相关性。方法:建立小鼠癫痫持续状态(SE);分别观察24 h和30 d,随机选取6只为急性致癫组,6只为慢性致癫组,通过行为学、电生理学及病理学HE染色方法验证模型的可靠性,建立人类颞叶癫痫小鼠模型。采用逆转录PCR方法检测DYRK1A mRNA在小鼠脑组织中的表达水平。结果:SE诱发成功率为64.71%(22/34),SE后死亡率为36.36%(8/22),;自发癫痫发作出现率为47.06%(8/17)。正常组和致癫组的海马神经元HE染色改变有差异,慢性期和急性期小鼠模型的脑电监测变化不一致。6只正常对照组DYRK1A mRNA与β-actin比值为0.4830±0.1243;6只急性致癫组DYRK1A mRNA与β-actin比值为0.8883±0.0727;6只慢性致癫组DYRK1A mRNA与β-actin比值为0.7112±0.1216;三组中两两比较差异均有统计学意义(P<0.05)。DYRK1A mRNA在三组中的表达情况:急性致癫组>慢性致癫组>正常对照组。结论:氯化锂-匹罗卡品诱导昆明小鼠癫痫持续状态后,脑组织中DYRK1A表达量较正常对照组增加,DYRK1A在氯化锂-匹罗卡品致癫痫模型的发生发展过程中可能起着重要的作用。

Objective： To establish a mice model of human temporal lobe epilepsy, lithium chloride-piloearpine can be induced status epileptieus seizures in Kunming mice by intraperitoneal injection.The analysis of DYRK1A mRNA expression in mice＇s brain tissue is used to investigate whether DYRK1A participate in the formation of hippocampai sclerosis of intractable temporal lobe epilepsy.Method： Epileptic group were established status epilepticus （SE）by injecting a small dose of lithium ehloride-piloearpine repeatedly and intraperitonealy.We randomly selected 6 mice into acute epileptic group, 6 mice into chronic epileptic group. On the basis of behavioral observation, electrophysiologlcal , patholo^eal HE staining method, we verified the reliability of the model and established mice model of human temporal lobe epilepsy.Reverse transcription PCR method was used to detect DYRK1A mRNA expression levels in the brain tissue of model mice.Result： There were 34 Kunming mice, which were be induced into status epileptieus seizures by intraperitoneal injection of lithium ehloride-piloearpine.22 Kunming mice were established status epilepticus （SE） and reached Racine Standard IV-V level, 6 mice were randomly selected into acute epilepsy group; The remaining 8 mice were observed within 30 days and 6 mice were selected into chronic epilepsy group; 6 mice in the control group were intraperitonealy injected by saline.They were not induced into any seizure and didn＇t die in the next 30 days.In this study, a success rate of induced SE was 64.71%（22/34）, the mortality rate after SE was 36.36%（8/22）; spontaneous seizures occurred at the rate of 47.06%（8/17）.Pathology HE staining： in comparison with the control group, hippocampal neurons of epileptic group arranged irregularly and the gap between neurons increased, with cell swelling, degeneration, necrosis, disintegration, cell body pyknosis, smaller volume, cytoplasm condensed hyperehromatie, nucleus pycnosis, unclear nueleoli, splitting decomposition of dead cell nuclei and eytoplasm.EEG monitoring of epileptic mice models ： in the acute phase, the outbreak of the long-range sharp activities, sharp spike activity and slow irregular composite activity was prominent in the background EEG activity; in the chronic phase, scattered frequency activities slower than the background slow activity and epileptic discharge appeared.In 6 control group, DYRK1A mRNA and β -actin ratio was 0.4830 ± 0.1243 ; in 6 acute epileptic group, DYRK1A mRNA and β -actin ratio was 0.8883 ± 0.0727 ; in 6 chronic epileptic group, DYRK1A mRNA and 13-actin ratio was 0.7112 ± 0.1216 ; The ratio differences between three groups all were statistically significant（P〈0.05）.DYRK1A mRNA expression in three groups： acute cause epilepsy group〉chronic epilepsy group〉control group. Conclusion： Lithium chloride-pilocarpine can be used to induce status epilepticus（SE） in Kunming mice.In comparison with control group, DYRK1A expression in brain tissue of SE Kumming mice model increased.