戊四氮致痫后大鼠海马中Caspase3和Hsp70的动态表达变化
摘要
目的:癫痫是以脑神经元过度异常放电导致反复发作性和短暂性的中枢神经系统功能失常为特征的一种脑功能失调综合征,其发病机制仍不清楚。
癫痫可致脑损伤,细胞凋亡或称程序性细胞死亡启动一系列复杂的蛋白酶级联反应,是癫痫致脑损伤的一条重要途径。长时间癫痫反复发作后可出现脑内选择性的神经元凋亡坏死和海马硬化,一旦出现了海马硬化,则脑组织损伤及记忆功能障碍将不可逆。通过腹腔注射戊四氮(pentylenetrazol,PTZ)建立癫痫持续状态大鼠模型来观察大鼠海马组织的形态学改变,如采用Nissl染色法检测神经细胞丢失坏死情况;Timm染色法观察海马组织苔藓纤维出芽情况;电镜观察海马CA3区的超微结构;同时运用免疫组化法计算阳性细胞的平均光密度值;Western blot方法分别检测海马组织中caspase3和hsp70的表达情况。初步探讨了癫痫发作后caspase3介导的凋亡相关途径和Hsp70的动态表达情况,将有助于从分子水平上更有效的寻求癫痫的治疗靶点,及时进行干预并控制凋亡的发生,为减轻癫痫致脑损伤提供新方法。
方法:选取清洁级(SPF级)雄性Sprague-Dawley (SD)大鼠70只,实验起始体重(180±20)g, 12h光亮/黑暗条件、22℃-25℃环境温度,分笼喂养。实验动物随机分为癫痫持续状态组(status epilepticus,SE组)和对照组(normal control group,NC组)两个大组,SE组又分为癫痫发作后3h组、6h组、12h组、24h组,5组各14只。SE各组大鼠应用生理盐水溶解的1% PTZ溶液,首先按40 mg/kg腹腔注射,10 min后再给20 mg/kg,根据实验动物的发作情况再适量追加10 mg/kg,直至出现Ⅳ级或Ⅴ级发作为止。Racine将点燃分为六级评价标准,至今仍在实验中广泛应用。0级,无任何反应:Ⅰ级,湿狗样抖动,面部抽搐及咀嚼;Ⅱ级,颈部肌肉痉挛,表现为点头和/或甩尾;Ⅲ级,一侧前肢痉挛;Ⅳ级,双侧前肢痉挛伴站立;Ⅴ级,全身阵挛、失去平衡甚至跌倒。完全点燃的标准是达到Ⅳ级或Ⅴ级发作且持续30min以上。对照组注射同等容量的生理盐水。观察大鼠行为学改变,后将各组大鼠随机选取3只行Nissl染色和Timm染色法分别检测海马组织神经细胞丢失坏死情况和苔藓纤维出芽情况,3只用于电镜观察海马CA3区的超微结构,5只行免疫组化法计算阳性细胞的平均光密度值,3只用于Western blot法分别进行检测海马组织中caspase3和hsp70的表达情况。实验过程中因持续强直发作而抽搐死亡的大鼠应及时进行补充(已补充28只)。
结果:1动物模型制备SE组有效应用的大鼠70只,11只应用40mg/ kg、14只50mg/ kg、15只60mg/ kg、18只70mg/ kg、8只80mg/ kg、4只90mg/ kg PTZ即开始出现惊厥,主要表现为节律性点头、肌阵挛。随着PTZ每10min注射一次,很快出现四肢抽搐,进入全面性强直发作致失去姿势控制,前、后肢强烈收缩,并伴有呼吸抑制,口周发紫,持续时间长短不等。强直阶段过后便出现全面性强直-阵挛发作。NC组动物均无行为学改变。
2大鼠海马组织学改变
2.1 Nissl染色观察SE组可见神经元皱缩体积缩小,胞核深染,有核分裂及核溶解,具有凋亡的特征,多数胞浆呈空泡状,海马CA1区细胞排列散乱,细胞密度小,尼氏体减少,部分神经元丢失,24h组最明显,3h脑损伤最轻;对照组海马神经元形态结构完整,胞核淡染、胞浆染色清晰,CA1区细胞几乎呈均匀一致的紫蓝色,细胞密度大,尼氏体着蓝色,形态接近正常。
2.2 Timm染色法观察SE各亚组和对照组的CA3区苔藓纤维出芽评分仅0～1分。
2.3海马CA3区超微结构观察SE组海马CA3区可见神经元核膜边界不清、可有内陷,染色质有边集,线粒体数量减少或有肿胀,24h组最明显,3h组较轻;对照组神经元核膜清晰,线粒体完整,细胞器基本完好。
3脑组织中caspase3和hsp70表达情况SE后3h caspase3和hsp70在海马神经细胞胞浆中着色开始增多,阳性细胞率(P <0.01与对照组比较),并随时间延长其阳性细胞率相应性升高,24h明显增高(P <0.01)。对照组可见caspase3和hsp70在海马胞浆中呈现散在着色。
4大鼠海马中caspase3和hsp70的蛋白表达SE后3h caspase3和hsp70表达开始增高(P <0.05与对照组比较),24h明显增高(P <0.05)。对照组仅有少量的caspase3和hsp70表达。
结论:1腹腔注射戊四氮可成功诱导大鼠癫痫持续状态,是目前模拟全身强直-阵挛性癫痫发作的一种较为理想的动物模型。
2癫痫发作急性期虽然引起了脑组织不同程度的形态学变化,但发作24h之内未出现明显的苔藓纤维出芽改变,表明大鼠的记忆功能无明显障碍,此期若能及时终止发作可逆转脑损伤。
3癫痫发作可导致神经细胞发生凋亡,同时上调caspase3和hsp70的表达可能对脑组织在应激方面起了重要的保护角色。
Objectives:Epilepsy has been a common clinical syndrome of functional disorder,which is essentially an abnormality of cerebral function on account of the discharges of some cerebral neuron in over high excitement state.However, the mechanics of epilepsy is not known yet.
Epilepsy can lead to brain indured.Apoptosis or programmed cell death resulting from a series of complicate protease enzyme cascade,is an important pathway of brain injured casuing by epilepsy.As we all know,long time seizu- re can lead to cell apoptosis,necrosis of neuron and sclerosis of hippocamps. Rats were kindled by PTZ,and then contributed to status epilepticus (SE).The changes of behavior were observed after seizure and the situation of neurocyte loss or necrosis and mossy fiber budding were determined with Nissl staining and Timm staining,respectively. Meanwhile, ultramicrostructure of hippocamp CA3 area was observed with electron microscope method.The average optical density of positive cell was counted with immunohistochemistry. Besides, exp -ressions of caspase3 and hsp70 in hippocamp of rats were determined by me- ans of Western blot. To investigate the dynamic state changes of caspases3 and hsp70 in hippocamp of rats after seizure ,thus we can adopt intervention and control neurocyte apoptosis in time.What’s more,we can explore a new method to lessen the brain damaged of epilepsy by means of molecular level.
Methods: Choosing status epilepticus(SE) model by PTZ-induced thro- ugh intraperitoneal injection,70 mature male Sprague-Dawley(SD) rats(weight 180±20g) were separated into two big groups randomly:status epilepticus group and normal control group.In addition.Each SE group is divided into 3h,6h,12h and 24h subset groups,and each group includes14 rats. Rats were injected by PTZ 40mg/kg, 20mg/kg after 10 mins,10mg/ kg after 10 mins. According to Racine criterion,seizure intensity was divided into 6 grades: grade 0:any seizure was not observe;grade , a little tingling of beards and twitching face; gradeⅡ,nodding,chewing accompanying with twitching face or tail; gradeⅢ, raising one of a forelimb and clonus;gradeⅣ,standing acco- mpanying with bilateral forelimbs;gradeⅤ: standing accompanying with fall -ing down,generalized tonic-clonic seizure. The kindled standard is that seizur -es of gradeⅣand gradeⅤcan be observed successively,which can last for above 30 mins. The changes of behavior were observed after seizure, and the average optical density of positive cell was counted with immunohistochemi -stry. In addition,the situation of neurocyte loss or necrosis and mossy fiber budding were determined with Nissl staining and Timm staining, respectively .Meanwhile, ultramicrostructure of hippocamp CA3 was observed by means of electron microscope . Expressions of caspase3 and hsp70 in hippocamp of rats were determined by Western blot method.
Results: 1 Results of epilepticus model:SE group using 70 rats effectively, 11 rats use 40mg/kg, 14 use 50mg/ kg,15 use 60mg/ kg,18 use 70mg/ kg, 8 use 80mg/ kg, 4 use 90mg/ kg,they began to nod,wash face and twitching face or tail.Accompanied by injecting every 10 mins,their muscles began to clonus, then forelimbs began to uplift and tonic clonic seizure at last. However, NC group rats did not appear epileptic seizure.
2 Changes of histology in hippocampus
2.1 Nissl staining: In hippocampal CA1 area of the rats in SE group , there was the following apoptosis changes: shrinkage of neuron, anachromasis of nucleus,karyokinesis and karyolysis. Besides,most of kytoplasm appears vacuolus and disorder of arrangement,nissl body diminished,fract lost of neuron .This change is the most obvious in 24h,while the neuron was damaged slightly in 3h . In hippocampal CA1 area of the rats in NC group,there were intact neurous, light staining of nucleus ,distinct structure and affluent kytopla -sm,and the morphous of nissl body are similar to normal.
2.2 Timm staining: In hippocampal CA3 area of SE subset groups and NC group ,the score of mossy fiber budding are merely 0 to 1 score .
2.3 Observation of ultramicrostructure in hippocampal CA3: In hippocampal CA3 area of the rats in SE groups , a series of changes are as follows: obscure boundary of karyolemma and emboly at times,chromatin with edge, diminished the quantity of mitochondria or swellen outline. The changes are the most obvious in 24h,while the changes are slight in 3h .As for the NC group ,appearing distinct boundary of karyolemma,intact mitochondria and organelles.
3 Changes of caspase3 and hsp70 in hippocampus:The expression of caspase3 and hsp70 began to increase after 3h in SE group,and then increased the highest in 24h(P <0.01 compared with NC group).Obviously, the rate of positive cells have a tendency to heighten with prolong of time. The cells of caspase3 and hsp70 can be scattered seen in the kytoplasm of hippocampus in NC group.
4 The expression of caspase3 and hsp70 protein in hippocampus:The expression of caspase3 and hsp70 protein began to increase slightly at 3h after seizure and at 24 h after seizure significantly(P <0.05 compared with NC group).There is a small amount of caspase3 and hsp70 protein in NC group.
Conclusion: 1 PTZ provides an effective model of postseizure dysfunc- tioning by intraperitoneal injection,which is similar to human being of tonic clonic seizure.
2 Although there are different morphology changes of brain tissue during acute seizure, yet there are not appear evident changes of mossy fiber budding in 24h,which indicates that the memory functions of rats have not been influen -ced obviously.If some effective measures can be taken,the damages of brain tissue can be reversed in time.
3 Seizure can lead to neurocyte apoptosis and upregulate expression of caspase3 and hsp70 simultaneously. The result demonstrates that neurocyte apoptosis at acute stage of seizure might play an important role in stress defen -dence of brian tissue.
癫痫可致脑损伤,细胞凋亡或称程序性细胞死亡启动一系列复杂的蛋白酶级联反应,是癫痫致脑损伤的一条重要途径。长时间癫痫反复发作后可出现脑内选择性的神经元凋亡坏死和海马硬化,一旦出现了海马硬化,则脑组织损伤及记忆功能障碍将不可逆。通过腹腔注射戊四氮(pentylenetrazol,PTZ)建立癫痫持续状态大鼠模型来观察大鼠海马组织的形态学改变,如采用Nissl染色法检测神经细胞丢失坏死情况;Timm染色法观察海马组织苔藓纤维出芽情况;电镜观察海马CA3区的超微结构;同时运用免疫组化法计算阳性细胞的平均光密度值;Western blot方法分别检测海马组织中caspase3和hsp70的表达情况。初步探讨了癫痫发作后caspase3介导的凋亡相关途径和Hsp70的动态表达情况,将有助于从分子水平上更有效的寻求癫痫的治疗靶点,及时进行干预并控制凋亡的发生,为减轻癫痫致脑损伤提供新方法。
方法:选取清洁级(SPF级)雄性Sprague-Dawley (SD)大鼠70只,实验起始体重(180±20)g, 12h光亮/黑暗条件、22℃-25℃环境温度,分笼喂养。实验动物随机分为癫痫持续状态组(status epilepticus,SE组)和对照组(normal control group,NC组)两个大组,SE组又分为癫痫发作后3h组、6h组、12h组、24h组,5组各14只。SE各组大鼠应用生理盐水溶解的1% PTZ溶液,首先按40 mg/kg腹腔注射,10 min后再给20 mg/kg,根据实验动物的发作情况再适量追加10 mg/kg,直至出现Ⅳ级或Ⅴ级发作为止。Racine将点燃分为六级评价标准,至今仍在实验中广泛应用。0级,无任何反应:Ⅰ级,湿狗样抖动,面部抽搐及咀嚼;Ⅱ级,颈部肌肉痉挛,表现为点头和/或甩尾;Ⅲ级,一侧前肢痉挛;Ⅳ级,双侧前肢痉挛伴站立;Ⅴ级,全身阵挛、失去平衡甚至跌倒。完全点燃的标准是达到Ⅳ级或Ⅴ级发作且持续30min以上。对照组注射同等容量的生理盐水。观察大鼠行为学改变,后将各组大鼠随机选取3只行Nissl染色和Timm染色法分别检测海马组织神经细胞丢失坏死情况和苔藓纤维出芽情况,3只用于电镜观察海马CA3区的超微结构,5只行免疫组化法计算阳性细胞的平均光密度值,3只用于Western blot法分别进行检测海马组织中caspase3和hsp70的表达情况。实验过程中因持续强直发作而抽搐死亡的大鼠应及时进行补充(已补充28只)。
结果:1动物模型制备SE组有效应用的大鼠70只,11只应用40mg/ kg、14只50mg/ kg、15只60mg/ kg、18只70mg/ kg、8只80mg/ kg、4只90mg/ kg PTZ即开始出现惊厥,主要表现为节律性点头、肌阵挛。随着PTZ每10min注射一次,很快出现四肢抽搐,进入全面性强直发作致失去姿势控制,前、后肢强烈收缩,并伴有呼吸抑制,口周发紫,持续时间长短不等。强直阶段过后便出现全面性强直-阵挛发作。NC组动物均无行为学改变。
2大鼠海马组织学改变
2.1 Nissl染色观察SE组可见神经元皱缩体积缩小,胞核深染,有核分裂及核溶解,具有凋亡的特征,多数胞浆呈空泡状,海马CA1区细胞排列散乱,细胞密度小,尼氏体减少,部分神经元丢失,24h组最明显,3h脑损伤最轻;对照组海马神经元形态结构完整,胞核淡染、胞浆染色清晰,CA1区细胞几乎呈均匀一致的紫蓝色,细胞密度大,尼氏体着蓝色,形态接近正常。
2.2 Timm染色法观察SE各亚组和对照组的CA3区苔藓纤维出芽评分仅0～1分。
2.3海马CA3区超微结构观察SE组海马CA3区可见神经元核膜边界不清、可有内陷,染色质有边集,线粒体数量减少或有肿胀,24h组最明显,3h组较轻;对照组神经元核膜清晰,线粒体完整,细胞器基本完好。
3脑组织中caspase3和hsp70表达情况SE后3h caspase3和hsp70在海马神经细胞胞浆中着色开始增多,阳性细胞率(P <0.01与对照组比较),并随时间延长其阳性细胞率相应性升高,24h明显增高(P <0.01)。对照组可见caspase3和hsp70在海马胞浆中呈现散在着色。
4大鼠海马中caspase3和hsp70的蛋白表达SE后3h caspase3和hsp70表达开始增高(P <0.05与对照组比较),24h明显增高(P <0.05)。对照组仅有少量的caspase3和hsp70表达。
结论:1腹腔注射戊四氮可成功诱导大鼠癫痫持续状态,是目前模拟全身强直-阵挛性癫痫发作的一种较为理想的动物模型。
2癫痫发作急性期虽然引起了脑组织不同程度的形态学变化,但发作24h之内未出现明显的苔藓纤维出芽改变,表明大鼠的记忆功能无明显障碍,此期若能及时终止发作可逆转脑损伤。
3癫痫发作可导致神经细胞发生凋亡,同时上调caspase3和hsp70的表达可能对脑组织在应激方面起了重要的保护角色。
Objectives:Epilepsy has been a common clinical syndrome of functional disorder,which is essentially an abnormality of cerebral function on account of the discharges of some cerebral neuron in over high excitement state.However, the mechanics of epilepsy is not known yet.
Epilepsy can lead to brain indured.Apoptosis or programmed cell death resulting from a series of complicate protease enzyme cascade,is an important pathway of brain injured casuing by epilepsy.As we all know,long time seizu- re can lead to cell apoptosis,necrosis of neuron and sclerosis of hippocamps. Rats were kindled by PTZ,and then contributed to status epilepticus (SE).The changes of behavior were observed after seizure and the situation of neurocyte loss or necrosis and mossy fiber budding were determined with Nissl staining and Timm staining,respectively. Meanwhile, ultramicrostructure of hippocamp CA3 area was observed with electron microscope method.The average optical density of positive cell was counted with immunohistochemistry. Besides, exp -ressions of caspase3 and hsp70 in hippocamp of rats were determined by me- ans of Western blot. To investigate the dynamic state changes of caspases3 and hsp70 in hippocamp of rats after seizure ,thus we can adopt intervention and control neurocyte apoptosis in time.What’s more,we can explore a new method to lessen the brain damaged of epilepsy by means of molecular level.
Methods: Choosing status epilepticus(SE) model by PTZ-induced thro- ugh intraperitoneal injection,70 mature male Sprague-Dawley(SD) rats(weight 180±20g) were separated into two big groups randomly:status epilepticus group and normal control group.In addition.Each SE group is divided into 3h,6h,12h and 24h subset groups,and each group includes14 rats. Rats were injected by PTZ 40mg/kg, 20mg/kg after 10 mins,10mg/ kg after 10 mins. According to Racine criterion,seizure intensity was divided into 6 grades: grade 0:any seizure was not observe;grade , a little tingling of beards and twitching face; gradeⅡ,nodding,chewing accompanying with twitching face or tail; gradeⅢ, raising one of a forelimb and clonus;gradeⅣ,standing acco- mpanying with bilateral forelimbs;gradeⅤ: standing accompanying with fall -ing down,generalized tonic-clonic seizure. The kindled standard is that seizur -es of gradeⅣand gradeⅤcan be observed successively,which can last for above 30 mins. The changes of behavior were observed after seizure, and the average optical density of positive cell was counted with immunohistochemi -stry. In addition,the situation of neurocyte loss or necrosis and mossy fiber budding were determined with Nissl staining and Timm staining, respectively .Meanwhile, ultramicrostructure of hippocamp CA3 was observed by means of electron microscope . Expressions of caspase3 and hsp70 in hippocamp of rats were determined by Western blot method.
Results: 1 Results of epilepticus model:SE group using 70 rats effectively, 11 rats use 40mg/kg, 14 use 50mg/ kg,15 use 60mg/ kg,18 use 70mg/ kg, 8 use 80mg/ kg, 4 use 90mg/ kg,they began to nod,wash face and twitching face or tail.Accompanied by injecting every 10 mins,their muscles began to clonus, then forelimbs began to uplift and tonic clonic seizure at last. However, NC group rats did not appear epileptic seizure.
2 Changes of histology in hippocampus
2.1 Nissl staining: In hippocampal CA1 area of the rats in SE group , there was the following apoptosis changes: shrinkage of neuron, anachromasis of nucleus,karyokinesis and karyolysis. Besides,most of kytoplasm appears vacuolus and disorder of arrangement,nissl body diminished,fract lost of neuron .This change is the most obvious in 24h,while the neuron was damaged slightly in 3h . In hippocampal CA1 area of the rats in NC group,there were intact neurous, light staining of nucleus ,distinct structure and affluent kytopla -sm,and the morphous of nissl body are similar to normal.
2.2 Timm staining: In hippocampal CA3 area of SE subset groups and NC group ,the score of mossy fiber budding are merely 0 to 1 score .
2.3 Observation of ultramicrostructure in hippocampal CA3: In hippocampal CA3 area of the rats in SE groups , a series of changes are as follows: obscure boundary of karyolemma and emboly at times,chromatin with edge, diminished the quantity of mitochondria or swellen outline. The changes are the most obvious in 24h,while the changes are slight in 3h .As for the NC group ,appearing distinct boundary of karyolemma,intact mitochondria and organelles.
3 Changes of caspase3 and hsp70 in hippocampus:The expression of caspase3 and hsp70 began to increase after 3h in SE group,and then increased the highest in 24h(P <0.01 compared with NC group).Obviously, the rate of positive cells have a tendency to heighten with prolong of time. The cells of caspase3 and hsp70 can be scattered seen in the kytoplasm of hippocampus in NC group.
4 The expression of caspase3 and hsp70 protein in hippocampus:The expression of caspase3 and hsp70 protein began to increase slightly at 3h after seizure and at 24 h after seizure significantly(P <0.05 compared with NC group).There is a small amount of caspase3 and hsp70 protein in NC group.
Conclusion: 1 PTZ provides an effective model of postseizure dysfunc- tioning by intraperitoneal injection,which is similar to human being of tonic clonic seizure.
2 Although there are different morphology changes of brain tissue during acute seizure, yet there are not appear evident changes of mossy fiber budding in 24h,which indicates that the memory functions of rats have not been influen -ced obviously.If some effective measures can be taken,the damages of brain tissue can be reversed in time.
3 Seizure can lead to neurocyte apoptosis and upregulate expression of caspase3 and hsp70 simultaneously. The result demonstrates that neurocyte apoptosis at acute stage of seizure might play an important role in stress defen -dence of brian tissue.
引文
1 Takaya S,Huanakawa T,Hashikawa K,et al.Prefrontalhypo function in patients with intractable mesial temporal lobe epilepsy.Neurology, 2006,67(9):1674-1676
2 Mortazavi F,Ericson M,Story D,et al.Spatial learning deficits and emotio- nal impairments in pentylenetetrazole-kindled rats. Epilepsy Behav, 2005, 7(4):629-638
3王维平,娄燕,李攀,等.癫痫后学习记忆障碍大鼠海马中脑源性神经营养因子表达的变化.第二军医大学学报,2007,28(5):488-491
4段瑞生,王维平,刘娜,等. SE大鼠认知、情感障碍与海马乙酰胆碱酯酶纤维分布改变的关系.河北医科大学学报,2007,28(1):15-18
5张映琦,廖维宏,迟路湘等.匹罗卡品致大鼠癫痫持续状态后海马神经元凋亡的动态观察.第三军医大学学报,2007,29(1):71-73
6 Qu H,Eloqayli H,Sonnewald U.Pentylenetetrazole affects meta- bolism of astrocytes in culture.J Neurosci Res,2005,79(1):48- 54
7 Fernandes Alnemri T ,Litwack G,Alnemri ES. CPP32 , a novel human apo -ptotic protein with homology to Caenorhabditis elegans cell death protein Ced23 and mammalian interleukin-1 beta-convert ing enzyme.J Biol Che -m,1994,269(49):30761-30764
8 Waterhouse N ,Kumar S ,Song Q ,et al.Heteronuclear ribonucle oproteins C1 and C2 ,components of the spliceosome,are specific targets of interleu- kin-1 beta converting enzyme like proteases in apoptosis.J Biol Chem, 1996,271(46):29335-29341
9 Sheng-jun Wang,Shu-hua Wang,Zhao-feng Song,et al.Poly(ADP- ribose) polymerase inhibitor is neuroprotective in epileptic rat via apoptosis indu -cing factor and Akt Signaling. Neuroreport,2007,18(12):1285-1289
10 Gillardon F ,Bottiger B ,Schmitz B ,et al . Activation of CPP232 protease in hippocampal neurons following ischemia and epilepsy. Brain Res Mol Brain Res,1997,50(22):16-22
11 Wang L,Liu YH,Huang YG,et al.Time-course of neuronal death in the mouse pilocarpine model of chronic epilepsy using FluoroJade C staining. Brain Res. 2008,124(1):157-167
12 Selvakumar P,Lakshmikuttyamma A,Charavaryamath C,et al.Expre ssion of myristoyltransferase and its interacting proteins in epilepsy. Biochem Biophys Res Commun. 2005,335(4):1132-1139
13郑虹,耿明.Hsp70、caspase3在胶质细胞瘤中的表达及与肿瘤病理分级的关系.山东大学学报,2007,4(59):944-946
14孙建英,关新华,张秀清,等.两种给药途径致痫大鼠海马神经元超微结构损伤及caspase3的表达特征.神经解剖学杂志,2007,23(6): 565- 570
1 Ebert U,Loscher W.Charaterization of phenytoin-resistant kindled rats,a new models of drug resistant partial epilepsy: influence of genetic factors.Epilepsy Res,1999,33(2-3): 217- 226
2张晓霞.电刺激复制癫痫病动物模型及其病理机制.实用临床医学, 2004,5(5):149-151
3 Matagne A,Klitgaard H.Validation of corneally kindled mice: a sensitive screening model for partial epilepsy in man.Epilepsy Res.1998,31 (1):59-71
4 Hesdorffer DC, Logroscino G, Cascino G,et al.Risk of unprovoked seizure after acute symptomatic seizure:effects of status epilepticus.Ann Neurol,1998,44(6):908-912
5 McIntyre DC,Gilby KL.Genetically seizure-prone or seizure sistant phenotypes and their associated behavioral comorbidities. Epilepsia. 2007,48(9):30-32
6陈英辉,赵永波,祖衡兵,等.杏仁核点燃癫大鼠脑内P-糖蛋白的表达.复旦学报,2007,34(4):568-571
7刘学伍,迟兆富.双侧杏仁核同时点燃建立复杂部分性癫痫模型的实验研究.临床神经病学杂志,2005,18(3):201-203
8张东君,周盛年.电刺激大鼠海马建立复杂部分性癫痫动物模型.中国神经科学杂志,2004,20(6):446-449
9陈启雄,蔡方成.戊四氮癫痫模型失神发作期脑电图及意识状态的实验研究.第三军医大学报,2007,29(4):335-337
10王超,张华,陈军,等.戊四氮对大鼠空间学习能力及情感行为的影响.立体定向和功能性神经外科杂志,2007,20(1):5-9
11张辉,宫瑾,隋鸿锦.急性癫痫发作后海马齿状回颗粒细胞的树突改变.解剖学杂志,2004,27(1):66-69
12 Nassiri Asl M,Shariati Rad S,Zamansoltani F,et al.Antico- nvulsive effects of intracerebroventricular administration of rutin in rats.Prog Neuropsychopharmacol Biol Psychiatry, 2008,32(4):989-993
13 Kang TC,Kang JH,Kim HT,et al.Anticonvulsant characteristics of pyridoxylgamma-aminobutyrate,PL-GABA.Neuropharmacology.2008, 54(6):954-964
14 Qu H, Eloqayli H, Sonnewald U. Pentylenetetrazole affects metabolism of astrocytesin culture.Neurosci Res,2005,79(1-2) :48-54
15 Erbayat Altay E,Yamada KA,Wong M,et al.Increased severity of pentylenetetyazol induced Seizures in leptin deficient ob/ob mice. Neurosci Lett,2008,433(2):82-86
16 Obay BD,Tasdemir E,Tumer C,et al.Dose dependent effects of ghrelin on pentylenetetrazole-induced oxidative stress in a rat seizure model. Pept- ides,2008,29(3):448-455
17 De Smedt T,De Rouck S,Raedt R,et al.Serial day rapid kindling is an effective tool in screening the anti-epileptic properties of topiramate. Seizure, 2007,16(7):620-626
18 Zhang JL,Zhang SP,Zhang HQ.Antiepileptic effects of electroacupunc–ture vs vagus nerve stimulation on cortical epileptiform activities. Neurol Sci,2008,270(1-2):114-121
19 Arida RM;Scorza FA;Scorza CA,et al. Is physical activity beneficial for recovery in temporal lobe epilepsy? Evidences from animal studies. Neurosci Biobehav Rev.2009,33(3): 422- 431
20朱遂强,杨嘉君,王开颜,等.戊四氮点燃过程中大鼠海马胶质细胞增生和突触重建的研究.中国临床神经外科杂志,2003,8(6):414- 41
2 Mortazavi F,Ericson M,Story D,et al.Spatial learning deficits and emotio- nal impairments in pentylenetetrazole-kindled rats. Epilepsy Behav, 2005, 7(4):629-638
3王维平,娄燕,李攀,等.癫痫后学习记忆障碍大鼠海马中脑源性神经营养因子表达的变化.第二军医大学学报,2007,28(5):488-491
4段瑞生,王维平,刘娜,等. SE大鼠认知、情感障碍与海马乙酰胆碱酯酶纤维分布改变的关系.河北医科大学学报,2007,28(1):15-18
5张映琦,廖维宏,迟路湘等.匹罗卡品致大鼠癫痫持续状态后海马神经元凋亡的动态观察.第三军医大学学报,2007,29(1):71-73
6 Qu H,Eloqayli H,Sonnewald U.Pentylenetetrazole affects meta- bolism of astrocytes in culture.J Neurosci Res,2005,79(1):48- 54
7 Fernandes Alnemri T ,Litwack G,Alnemri ES. CPP32 , a novel human apo -ptotic protein with homology to Caenorhabditis elegans cell death protein Ced23 and mammalian interleukin-1 beta-convert ing enzyme.J Biol Che -m,1994,269(49):30761-30764
8 Waterhouse N ,Kumar S ,Song Q ,et al.Heteronuclear ribonucle oproteins C1 and C2 ,components of the spliceosome,are specific targets of interleu- kin-1 beta converting enzyme like proteases in apoptosis.J Biol Chem, 1996,271(46):29335-29341
9 Sheng-jun Wang,Shu-hua Wang,Zhao-feng Song,et al.Poly(ADP- ribose) polymerase inhibitor is neuroprotective in epileptic rat via apoptosis indu -cing factor and Akt Signaling. Neuroreport,2007,18(12):1285-1289
10 Gillardon F ,Bottiger B ,Schmitz B ,et al . Activation of CPP232 protease in hippocampal neurons following ischemia and epilepsy. Brain Res Mol Brain Res,1997,50(22):16-22
11 Wang L,Liu YH,Huang YG,et al.Time-course of neuronal death in the mouse pilocarpine model of chronic epilepsy using FluoroJade C staining. Brain Res. 2008,124(1):157-167
12 Selvakumar P,Lakshmikuttyamma A,Charavaryamath C,et al.Expre ssion of myristoyltransferase and its interacting proteins in epilepsy. Biochem Biophys Res Commun. 2005,335(4):1132-1139
13郑虹,耿明.Hsp70、caspase3在胶质细胞瘤中的表达及与肿瘤病理分级的关系.山东大学学报,2007,4(59):944-946
14孙建英,关新华,张秀清,等.两种给药途径致痫大鼠海马神经元超微结构损伤及caspase3的表达特征.神经解剖学杂志,2007,23(6): 565- 570
1 Ebert U,Loscher W.Charaterization of phenytoin-resistant kindled rats,a new models of drug resistant partial epilepsy: influence of genetic factors.Epilepsy Res,1999,33(2-3): 217- 226
2张晓霞.电刺激复制癫痫病动物模型及其病理机制.实用临床医学, 2004,5(5):149-151
3 Matagne A,Klitgaard H.Validation of corneally kindled mice: a sensitive screening model for partial epilepsy in man.Epilepsy Res.1998,31 (1):59-71
4 Hesdorffer DC, Logroscino G, Cascino G,et al.Risk of unprovoked seizure after acute symptomatic seizure:effects of status epilepticus.Ann Neurol,1998,44(6):908-912
5 McIntyre DC,Gilby KL.Genetically seizure-prone or seizure sistant phenotypes and their associated behavioral comorbidities. Epilepsia. 2007,48(9):30-32
6陈英辉,赵永波,祖衡兵,等.杏仁核点燃癫大鼠脑内P-糖蛋白的表达.复旦学报,2007,34(4):568-571
7刘学伍,迟兆富.双侧杏仁核同时点燃建立复杂部分性癫痫模型的实验研究.临床神经病学杂志,2005,18(3):201-203
8张东君,周盛年.电刺激大鼠海马建立复杂部分性癫痫动物模型.中国神经科学杂志,2004,20(6):446-449
9陈启雄,蔡方成.戊四氮癫痫模型失神发作期脑电图及意识状态的实验研究.第三军医大学报,2007,29(4):335-337
10王超,张华,陈军,等.戊四氮对大鼠空间学习能力及情感行为的影响.立体定向和功能性神经外科杂志,2007,20(1):5-9
11张辉,宫瑾,隋鸿锦.急性癫痫发作后海马齿状回颗粒细胞的树突改变.解剖学杂志,2004,27(1):66-69
12 Nassiri Asl M,Shariati Rad S,Zamansoltani F,et al.Antico- nvulsive effects of intracerebroventricular administration of rutin in rats.Prog Neuropsychopharmacol Biol Psychiatry, 2008,32(4):989-993
13 Kang TC,Kang JH,Kim HT,et al.Anticonvulsant characteristics of pyridoxylgamma-aminobutyrate,PL-GABA.Neuropharmacology.2008, 54(6):954-964
14 Qu H, Eloqayli H, Sonnewald U. Pentylenetetrazole affects metabolism of astrocytesin culture.Neurosci Res,2005,79(1-2) :48-54
15 Erbayat Altay E,Yamada KA,Wong M,et al.Increased severity of pentylenetetyazol induced Seizures in leptin deficient ob/ob mice. Neurosci Lett,2008,433(2):82-86
16 Obay BD,Tasdemir E,Tumer C,et al.Dose dependent effects of ghrelin on pentylenetetrazole-induced oxidative stress in a rat seizure model. Pept- ides,2008,29(3):448-455
17 De Smedt T,De Rouck S,Raedt R,et al.Serial day rapid kindling is an effective tool in screening the anti-epileptic properties of topiramate. Seizure, 2007,16(7):620-626
18 Zhang JL,Zhang SP,Zhang HQ.Antiepileptic effects of electroacupunc–ture vs vagus nerve stimulation on cortical epileptiform activities. Neurol Sci,2008,270(1-2):114-121
19 Arida RM;Scorza FA;Scorza CA,et al. Is physical activity beneficial for recovery in temporal lobe epilepsy? Evidences from animal studies. Neurosci Biobehav Rev.2009,33(3): 422- 431
20朱遂强,杨嘉君,王开颜,等.戊四氮点燃过程中大鼠海马胶质细胞增生和突触重建的研究.中国临床神经外科杂志,2003,8(6):414- 41