癫痫持续发作时间与大鼠海马苔藓纤维发芽程度及自发性痫性发作的关系
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摘要
目的探讨癫痫持续状态(SE)发作时间与致痫大鼠海马苔藓纤维发芽(MFS)程度及自发性痫性发作的关系。方法 104只雄性成年SD大鼠,随机分为对照组和3个SE实验组,建立氯化锂-重复低剂量匹罗卡品致痫大鼠模型;诱发SE30min(A组)、60min(B组)、90min(C组)后注射水合氯醛终止发作。各组大鼠自SE终止发作后于相同实验条件下普通饲养45d,观察大鼠行为及脑电图(EEG)的变化,记录自发性痫性发作的发生率。通过苏木精-伊红染色、Nissl染色和Timm硫化银组织化学染色方法观察各实验组海马MFS情况。结果氯化锂-重复低剂量匹罗卡品成功诱导大鼠SE的发生,发作程度均达Ⅳ级以上,EEG类似人类颞叶癫痫。80%的大鼠癫痫持续状态均发展为自发痫性发作,与SE时间无关。与对照组相比,实验A、B、C三组双侧海马CA3区均表现MFS(P<0.05)。实验B组与A、C组相比,CA3区MFS明显增加(P<0.05)。结论氯化锂-重复低剂量匹罗卡品可诱导SE,癫痫持续发作60min后终止的大鼠海马CA3区MFS明显增加,SE发作时间与海马MFS程度并不一定呈正相关。
Objective To investigate the relationship between the onset time of status epilepticus(SE) and the degree of mossy fiber sprouting(MFS) in hippocampal and spontaneous seizures in rats with epilepsy. Method Totally 104 male adult healthy Sprague-Dawley(SD) rats were randomly divided into control group and epilepsy continuous state model groups to establish epilepsy rat model with lithium chloride-repeated low-dose pilocarpine. In the experimental group, the onset of status epilepticus 30 minutes(group A), 60 minutes(group B) and 90 minutes(group C) was terminated by hydrated chloral injection. The rats in each group were raised for 45 days under the same experimental conditions after status epilepticus termination, and the changes of behavior and electroencephalogram(EEG) were observed and the incidence of spontaneous seizures was recorded. The hippocampal mossy fiber sprouting of each experimental group were observed by Hematoxylin-eosin staining, Nissl staining and Timm's silver sulfide histochemistry staining. Results Lithium chloride-repeated low-dose pirocapine can induce status epilepticus, the extent of Ⅳ level or above, and electroencephalogram is similar to human temporal lobe epilepsy. Eighty percent of the rats developed spontaneous epileptic seizures, regardless of the onset time of status epilepticus. Mossy fiber sprouting in CA3 area of bilateral hippocampus in the three groups of experiment A, B and C compared to the control group(P<0.05).Compared with group A and group B, the mossy fiber sprouting in CA3 area was significantly increased of experiment B(P<0.05). Conclusion Lithium chloride-repeated low-dose pilocarpine can induce status epilepticus,mossy fiber sprouting in the hippocampal CA3 area of rats terminated after 60 minutes of epileptic seizures increased significantly, and the onset time of status epilepticus was not necessarily positively correlated with hippocampal mossy fiber sprouting.
Objective To investigate the relationship between the onset time of status epilepticus(SE) and the degree of mossy fiber sprouting(MFS) in hippocampal and spontaneous seizures in rats with epilepsy. Method Totally 104 male adult healthy Sprague-Dawley(SD) rats were randomly divided into control group and epilepsy continuous state model groups to establish epilepsy rat model with lithium chloride-repeated low-dose pilocarpine. In the experimental group, the onset of status epilepticus 30 minutes(group A), 60 minutes(group B) and 90 minutes(group C) was terminated by hydrated chloral injection. The rats in each group were raised for 45 days under the same experimental conditions after status epilepticus termination, and the changes of behavior and electroencephalogram(EEG) were observed and the incidence of spontaneous seizures was recorded. The hippocampal mossy fiber sprouting of each experimental group were observed by Hematoxylin-eosin staining, Nissl staining and Timm's silver sulfide histochemistry staining. Results Lithium chloride-repeated low-dose pirocapine can induce status epilepticus, the extent of Ⅳ level or above, and electroencephalogram is similar to human temporal lobe epilepsy. Eighty percent of the rats developed spontaneous epileptic seizures, regardless of the onset time of status epilepticus. Mossy fiber sprouting in CA3 area of bilateral hippocampus in the three groups of experiment A, B and C compared to the control group(P<0.05).Compared with group A and group B, the mossy fiber sprouting in CA3 area was significantly increased of experiment B(P<0.05). Conclusion Lithium chloride-repeated low-dose pilocarpine can induce status epilepticus,mossy fiber sprouting in the hippocampal CA3 area of rats terminated after 60 minutes of epileptic seizures increased significantly, and the onset time of status epilepticus was not necessarily positively correlated with hippocampal mossy fiber sprouting.
引文
[1]Liu H,Yang Y,Wang Y,et al.Ketogenic diet for treatment of intractable epilepsy in adults:A meta-analysis of observational studies[J].Epilepsia Open,2018,3(1):9-17.
[2]Glien M,Brandt C,Potschka H,et al.Repeated low-dose treatment of rats with pilocarpine:low mortality but high proportion of rats developing epilepsy[J].Epilepsy Res,2001,46(2):111-119.
[3]Haussler U,Rinas K,Kilias A,et al.Mossy fiber sprouting and pyramidal cell dispersion in the hippocampal CA2 region in a mouse model of temporal lobe epilepsy[J].Hippocampus,2016,26(5):577-588.
[4]Racine R J.Modification of seizure activity by electrical stimulation.II.Motor seizure[J].Electroencephalogr Clin Neurophysiol,1 972,32(3):281-294.
[5]刘菁,李保敏,孙若鹏,等.幼鼠反复癫痫发作致苔藓纤维发芽的动态变化及其与自发性发作的关系[J].山东大学学报(医学版),2013,51(11):16-20.
[6]Heng K,Haney M M,Buckmaster P S.High-dose rapamycin blocks mossy fiber sprouting but not seizures in a mouse model of temporal lobe epilepsy[J].Epilepsia,2013,54(9):1535-1541.
[7]Scharfman H E,Sollas A L,Berger R E,et al.Electrophysiological evidence of monosynaptic excitatory transmission between granule cells after seizure-induced mossy fiber sprouting[J].JNeurophysiol,2003,90(4):2536-2547.
[8]张小东,黄敏,许可,等.氯化锂-重复低剂量匹罗卡品致大鼠癫痫模型制作[J].重庆医科大学学报,2008,33(1):63-65.
[9]李丹,黄绍平,马伟军,等.氯化锂-匹鲁卡品癫痫动物模型的建立[J].陕西医学杂志,2006,35(1):15-17.
[10]Lemos T,Cavalheiro E A.Suppression of pilocarpine-induced status epilepticus and the late development of epilepsy in rats[J].Exp Brain Res,1995,102(3):423-428.
[11]Buckmaster P S,Lew F H.Rapamycin suppresses mossy fiber sprouting but not seizure frequency in a mouse model of temporal lobe epilepsy[J].J Neurosci,2011,31(6):2337-2347.
[12]Pitsch J,Becker A J,Schoch S,et al.Circadian clustering of spontaneous epileptic seizures emerges after pilocarpine-induced status epilepticus[J].Epilepsia,2017,58(7):1159-1171.
[13]Drexel M,Preidt A P,Sperk G.Sequel of spontaneous seizures after kainic acid-induced status epilepticus and associated neuropathological changes in the subiculum and entorhinal cortex[J].Neuropharmacology,2012,63(5):806-817.
[14]Joushi S,Salmani M E.Effect of eugenol on lithium-pilocarpine model of epilepsy:behavioral,histological,and molecular changes[J].Iran J Basic Med Sci,2017,20(7):745-752.
[15]Neumann A M,Abele J,Kirschstein T,et al.Mycophenolate mofetil prevents the delayed T cell response after pilocarpine-induced status epilepticus in mice[J].PLoS One,2017,12(11):e0187330.
[16]Lee S H,Lutz D,Mossalam M,et al.Presenilins regulate synaptic plasticity and mitochondrial calcium homeostasis in the hippocampal mossy fiber pathway[J].Mol Neurodegener,2017,12(1):48.
[2]Glien M,Brandt C,Potschka H,et al.Repeated low-dose treatment of rats with pilocarpine:low mortality but high proportion of rats developing epilepsy[J].Epilepsy Res,2001,46(2):111-119.
[3]Haussler U,Rinas K,Kilias A,et al.Mossy fiber sprouting and pyramidal cell dispersion in the hippocampal CA2 region in a mouse model of temporal lobe epilepsy[J].Hippocampus,2016,26(5):577-588.
[4]Racine R J.Modification of seizure activity by electrical stimulation.II.Motor seizure[J].Electroencephalogr Clin Neurophysiol,1 972,32(3):281-294.
[5]刘菁,李保敏,孙若鹏,等.幼鼠反复癫痫发作致苔藓纤维发芽的动态变化及其与自发性发作的关系[J].山东大学学报(医学版),2013,51(11):16-20.
[6]Heng K,Haney M M,Buckmaster P S.High-dose rapamycin blocks mossy fiber sprouting but not seizures in a mouse model of temporal lobe epilepsy[J].Epilepsia,2013,54(9):1535-1541.
[7]Scharfman H E,Sollas A L,Berger R E,et al.Electrophysiological evidence of monosynaptic excitatory transmission between granule cells after seizure-induced mossy fiber sprouting[J].JNeurophysiol,2003,90(4):2536-2547.
[8]张小东,黄敏,许可,等.氯化锂-重复低剂量匹罗卡品致大鼠癫痫模型制作[J].重庆医科大学学报,2008,33(1):63-65.
[9]李丹,黄绍平,马伟军,等.氯化锂-匹鲁卡品癫痫动物模型的建立[J].陕西医学杂志,2006,35(1):15-17.
[10]Lemos T,Cavalheiro E A.Suppression of pilocarpine-induced status epilepticus and the late development of epilepsy in rats[J].Exp Brain Res,1995,102(3):423-428.
[11]Buckmaster P S,Lew F H.Rapamycin suppresses mossy fiber sprouting but not seizure frequency in a mouse model of temporal lobe epilepsy[J].J Neurosci,2011,31(6):2337-2347.
[12]Pitsch J,Becker A J,Schoch S,et al.Circadian clustering of spontaneous epileptic seizures emerges after pilocarpine-induced status epilepticus[J].Epilepsia,2017,58(7):1159-1171.
[13]Drexel M,Preidt A P,Sperk G.Sequel of spontaneous seizures after kainic acid-induced status epilepticus and associated neuropathological changes in the subiculum and entorhinal cortex[J].Neuropharmacology,2012,63(5):806-817.
[14]Joushi S,Salmani M E.Effect of eugenol on lithium-pilocarpine model of epilepsy:behavioral,histological,and molecular changes[J].Iran J Basic Med Sci,2017,20(7):745-752.
[15]Neumann A M,Abele J,Kirschstein T,et al.Mycophenolate mofetil prevents the delayed T cell response after pilocarpine-induced status epilepticus in mice[J].PLoS One,2017,12(11):e0187330.
[16]Lee S H,Lutz D,Mossalam M,et al.Presenilins regulate synaptic plasticity and mitochondrial calcium homeostasis in the hippocampal mossy fiber pathway[J].Mol Neurodegener,2017,12(1):48.