白藜芦醇调控癫痫持续状态大鼠大脑皮质内源性巯醇抗氧化剂(酶)的作用研究

目的:探讨白藜芦醇(resveratrol,Res)在氯化锂(LiCl)-重复低剂量匹罗卡品(pilocarpine,Pilo)诱导癫痫持续状态(status epilepticus,SE)大鼠模型中的抗氧化作用及其机制。方法:54只SD大鼠随机分为3组:对照组、癫痫模型组和Res治疗组。采用重复低剂量氯化锂-联合皮罗卡品腹腔注射制备癫痫持续状态SD大鼠模型,并同时给予Res预防性治疗,对实验动物按痫性分级标准进行行为学观测,采用化学比色法检测痫性发作90min后模型动物大脑皮质过氧化氢酶(catalase,CAT)、谷胱甘肽(glutathion,GSH)、超氧化物歧化酶(superoxidedismutase,SOD)和谷胱甘肽还原酶(glutathion reductase,GR)含量变化。采用免疫组织化学法和免疫印迹法(Western blot)检测痫性发作90 min后模型动物大脑皮质中凋亡相关蛋白Bcl-2、Caspase-3和Bax的表达变化。结果:癫痫大鼠大脑皮质CAT、GR、SOD和GSH的含量较对照组显著减少(P<0.05),凋亡相关蛋白Bcl-2表达下调,该蛋白阳性细胞数减少,而caspase-3和Bax表达显著上调(P<0.05),两种蛋白阳性细胞数增多(P<0.05);Res预防性治疗后,治疗组动物痫性发作潜伏期较模型组显著延长(P<0.05);药物治疗明显上调模型动物大脑皮质中巯醇抗氧化剂(酶)CAT、GR、SOD和GSH的含量(P<0.05),增加抗凋亡蛋白Bcl-2表达(P<0.05)和阳性细胞数(P<0.05),同时抑制促凋亡因子Caspase-3和Bax的表达(P<0.05)和减少两种蛋白阳性细胞数(P<0.05)。结论:Res能通过上调癫痫模型大鼠皮质内源性巯基抗氧化物(酶)CAT、GR、SOD和GSH的含量,调控凋亡相关蛋白Bcl-2、caspase-3和Bax的表达变化,对抗SE诱导的神经细胞氧化损伤,发挥神经保护作用。     

ASIC1a和TASKs在大鼠癫痫持续状态模型中的作用

目的:探讨酸敏感离子通道1a(ASIC1a)、弱内向整流钾通道相关的酸敏感钾通道1和3(TASK-1和TASK-3)在大鼠癫痫持续状态(SE)中的表达及其在癫痫持续状态中的作用。方法:成年雄性SD大鼠随机分为对照组(control)和SE组,应用氯化锂-匹罗卡品诱导SE模型,采用real time RT-PCR和Western Blot技术分别检测海马组织ASIC1a、TASK-1和TASK-3在SE后0、1、2和3 h mRNA、蛋白表达水平,应用膜片钳技术观察SE后ASIC1a、TASK-1和TASK-3对海马CA1区锥体神经元兴奋性的影响。结果:在mRNA水平,与control组相比,SE组ASIC1a在2和3 h、TASK-1在1 h、TASK-3在3 h时间点表达显著减少。在蛋白水平,与control组相比,SE组ASIC1a、TASK-1和TASK-3在3 h时间点表达均显著降低。与SE control组相比,给予ASIC1a抑制剂后动作电位(AP)的频率明显降低,给予TASK-1和TASK-3抑制剂后,AP的频率均显著增加。结论:SE后大鼠海马区ASIC1a、TASK-1...     

Opioid peptide pharmacology and immunocytochemistry in an animal model of self-sustaining status epilepticus

In a model of self-sustaining status epilepticus induced in rats by 30 min intermittent stimulation of the perforant path through chronically implanted electrodes, a decrease in dynorphin-like immunoreactivity in the dentate gyrus and CA3 was observed 3 h and 24 h after the induction of status epilepticus. Enkephalin-like immunoreactivity decreased 3 h but not 24 h after perforant path stimulation. Injection into the hilus of the dentate gyrus 10 min prior to stimulation of the kappa-receptor agonist dynorphin-A(1-13), the delta-receptor antagonists ICI-174864 and naltrindole, as well as i.p. injection of naloxone prevented the development of status epilepticus. Perihilar administration of the delta-agonist [D-Ser2]Leu-enkephalin-Thr6 or the kappa-antagonist nor-Binaltorphimine, but not of the mu-agonist [D-Ala2,N-Me-Phe4,Gly-ol5]-Enkephalin, facilitated the establishment of self-sustaining status epilepticus. Injection into the hilus of dynorphin-A(1-13) after the end of perforant path stimulation, stopped established status epilepticus, while administration of naloxone, naltrindole and ICI-174864 were ineffective. We conclude that kappa-opioids in the hippocampus counteract initiation and maintenance of status epilepticus, while delta-opioids promote initiation, but not maintenance of seizure activity. These data are important for the understanding the mechanisms which underlie initiation and maintenance of status epilepticus and for the development of new approaches for its effective management.     

Long-lasting induction of brain-derived neurotrophic factor is restricted to resistant cell populations in an animal model of status epilepticus

We have recently characterized an animal model of status epilepticus induced by a single intraseptal injection of kainate. Under these conditions, there is a delayed expanding apoptotic hippocampal and amygdalar cell death. In order to further characterize this animal model, we have performed a detailed time-course analysis of the appearance of cell death, brain-derived neurotrophic factor messenger RNA expression and astroglial and microglial response in different brain areas related to the limbic system. We found a long-lasting delayed apoptotic cell death in the hippocampal formation, amygdala, medial thalamus, dorsal endopiriform nucleus and multiple cortical areas from two to 21 days post-injection. There was a spatiotemporal correlation between the appearance of cell death and induction of brain-derived neurotrophic factor messenger RNA expression in the areas studied, and interestingly this induction was found in non-degenerating cells.We conclude that our animal model of status epilepticus exhibits remarkable features of recurrent seizure activity and provides evidence for a neuroprotective role of brain-derived neurotrophic factor against seizure-induced apoptotic cell death.     

40例癫（间）持续状态患者的临床特点及脑电变化特征

目的：分析不同类型癫痫持续状态（SE）患者的临床特点及脑电图（EEG）变化特征并观察临床疗效。方法：对40例SE患者进行临床分类并记录发作间期及发作期的EEG，静脉注射安定进行治疗并观察其临床疗效。结果：惊厥性SE28例，其中强直阵挛性SE24例，肌阵挛性发作SE1例，单纯部分运动性SE3例；非惊厥性SE12例，其中复杂部分性SE10例不典型失神SE2例，发作间期EEG25例记录到癫痫样放电；发作期EEG，8例病人行Video-EEG监测，7例记录到发作期典型癫痫样放电。1例表现为募集节律。疗效；治愈率95%，死亡率5%。结论：强直阵挛性SE常见，发作间期多可记录到癫痫样波形，肌阵挛性SE、非惊厥性SE临床诊断困难。应行Video-EEG监测；早期足量应用安定是安全有效的治疗措施。     

39例复杂部分性发作癫灶的PET与EEG定位诊断比较

目的 :探讨临床表现不同的癫复杂部分性发作 (CPS)与正电子发射断层扫描 (PET)检查病灶定位的相关性。方法 :39例CPS病人经PET、磁共振成像 (MRI)、录像 脑电监测 (Video EEG)检查 ,并对各种检查和临床表现进行比较分析。结果 :PET和EEG二者完全符合和部分符合均为 31% ,15例不符病例中 ,12例为PET有定位价值而EEG不能定位。仅表现为复杂部分性发作而无继发性全身性强直阵挛发作 (GS)者的 71%的病例 ,PET定位于单侧颞叶 ,表现为CPS继发GS者的 81%为颞顶、颞枕区病灶。结论 :CPS不继发全身性发作的患者 ,病灶多局限于一侧颞叶 ,如发作期和发作间期脑电图均正常 ,更应考虑病灶位于颞叶内侧面 ,或边缘系统 ;CPS继发GS的患者病灶多位于颞叶外侧面与顶、枕交界区。     

Editorial commentary on special issue of Advances in EEG Signal Processing and Machine Learning for Epileptic Seizure Detection and Prediction

This special issue of The Journal of Biomedical Research features novel studies on epileptic seizure detection and prediction based on advanced EEG signal processing and machine learning algorithms. The articles selected present important findings including new experimental results and theoretical studies.     

Age-dependent mortality in the pilocarpine model of status epilepticus

Status epilepticus (SE) is an acute neurological emergency associated with significant morbidity and mortality. Age has been shown to be a critical factor in determining outcome after SE. Understanding the causes of this increased mortality with aging by developing an animal model to study this condition would play a major role in studying mechanisms to limit the mortality due to SE. Here we employed pilocarpine to induce SE in rats aged between 5 and 28 weeks. Similar to clinical studies in man, we observed that age was a significant predictor of mortality following SE. While no deaths were observed in 5-week-old animals, mortality due to SE increased progressively with age and reached 90% in 28-week-old animals. There was no correlation between the age of animals and severity of SE. With increasing age mortality occurred earlier after the onset of SE. These results indicate that pilocarpine-induced SE in the rat provides a useful model to study age-dependent SE-induced mortality and indicates the importance of using animal models to elucidate the mechanisms contributing to SE-induced mortality and the development of novel therapeutic interventions to prevent SE-induced death.     

海人酸致癫痫大鼠海马结构中noggin表达变化

目的:研究海人酸(kainic acid,KA)侧脑室注射并诱发癫痫持续状态(status epilepticus,SE)后大鼠海马结构中noggin基因在大鼠海马的表达变化。方法:大鼠在侧脑室注射KA后1、3、7、14、30和60d等不同时间,采用RT-PCR研究noggin mRNA含量变化,采用免疫组化观察noggin蛋白表达变化。结果:在正常大鼠海马结构中noggin mRNA有少量表达。noggin阳性细胞主要位于齿状回及CA3、CA1区,数量较少。侧脑室注射KA诱发SE后,noggin表达持续升高,3d达到高峰;7d在脑内的表达开始降低;注射后2个月,noggin表达降至术前水平,仅见散在的阳性细胞。结论:侧脑室注射KA并诱发SE后,大鼠海马结构中noggin表达明显增加。     

Allergenicity of recombinant human lactoferrin to an animal model Brown Norway rats

The assessment of allergenicity of recombinant human lactoferrin (rhLF) using an animal model for its extensive use. In this study, 32 Brown Norway (BN) rats were divided into four groups that were exposed to ovalbumin (OVA) (positive control), rhLF, bovine lactoferrin (bLF) and saline (negative control), respectively, with daily intragastric administration for 42 days. The level of specific antibody (IgE, IgG and IgG2a), blood cell counts, blood pressure and pathology of important organs were measured and compared among the groups. OVA stimulated significant higher levels of specific-IgE and eosinophil (EO) counts (p <0.05), lower blood pressure (p<0.05) and more serious lesion in the lung of BN rats compared to the other groups. And the animals did not show obvious humeral, cell-mediated and local allergic response to rhLF and bLF. The results demonstrate a weak allergenicity of rhLF on the animal and reveal its potential value for further identified study.     

The immunosuppressant cyclosporin A inhibits recurrent seizures in an experimental model of temporal lobe epilepsy

The immunosuppressant, cyclosporin A (CsA), is neuroprotective following brain injury. Previous studies suggest that CsA treatment ameliorates seizure severity during status epilepticus (SE) or cell death following SE. The antiepileptic effects of CsA on recurrent seizures, however, have not been investigated. In the present study, the effects of CsA on spontaneous recurrent seizures (SRSs) in a kainate (KA)-induced mouse model of mesial temporal lobe epilepsy (TLE) were examined. Moreover, the effects of CsA on epileptiform activity in a 4-aminopyridine (4-AP)-induced in vitro seizure model were investigated. A mesial TLE mouse model was generated with a unilateral intrahippocampal injection of KA. SRSs were determined in the ipsilateral hippocampal CA1 region with a long-term video-EEG. CsA was systemically administrated to the epileptic mice exhibiting a stable occurrence of SRSs. A 1-mg/kg dose of CsA did not have any effect on SRSs in the epileptic mice. However, a 5-mg/kg dose of CsA significantly reduced the number of SRSs and decreased the severity of the seizures in the epileptic mice. Additionally, CsA treatment inhibited spontaneous burst discharges in 4-AP-treated hippocampal slices. The results of the present study demonstrate that CsA inhibits recurrent seizures in a mouse model of mesial TLE and suggest that CsA may afford both neuroprotection against SE and antiepileptic effects during the chronic period of epilepsy.     

Postmenopausal effects of intrastriatal estrogen on catalepsy and pallidal electroencephalogram in an animal model of Parkinson's disease

The main objective was to test the preventive and treatment effects of central injection of estrogen (ES) on muscular rigidity and pallidal EEG in menopausal rats' model of Parkinson's disease (PD). We hypothesized that intrastriatal pretreatment and post-lesion treatment by estrogen improve the pallidal local EEG and muscular stiffness in animal model of menopause with PD. Forty-eight female Wistar rats (300-350 g) were ovariectomized (OVX) and divided into two main groups: Non-pretreated subgroups; sham (S), lesioned (L), post-lesion treated (LT) and pretreated subgroups; pretreated (Pt), pretreated and then lesioned (PtL), pretreated and post-lesion treated (PtLT). Pallidal local EEG was recorded by a bipolar recording electrode and muscle stiffness was scored by Dekundy's test in freely moving rats. Muscle stiffness and pallidal local EEG were indicated as main outcome measures. In pretreated group the local pallidal EEG was decreased in sham-operated rats compared with non-pretreated group (P<0.01), and SNc lesioning decreased EEG in the non-pretreated (P<0.01), while it increased the EEG in the pretreated group (P<0.01). In both groups administration of ES restore the EEG to the respective sham-operated group (P<0.01). Regarding muscle stiffness, it increased after SNc lesioning in both pretreated and non-pretreated groups and ES administration decreased the rigidity significantly (P<0.05, P<0.001 respectively). However, the lesion-induced rigidity in pretreated groups was significantly less than non-pretreated groups (P<0.05). Because of its modulatory effect estrogen may protect dopaminergic neurons from injury and may interfere with the uptake of 6-hydroxydopamine (6-OHDA) into the nigral dopaminergic neurons or alter dopamine release and uptake in remaining neurons.     

Abortive amygdaloid kindled seizures following micro-injection of γ-vinyl-GABA in the vicinity of substantia nigra in rats

The effects of microinjection of a GABA-elevating substance (γ-vinyl-GABA) in the substantia nigra were assessed on kindled convulsive seizures induced by daily appropriate amygdaloid stimulation in the rat. Bilateral administration of 20 μg of γ-vinyl-GABA strongly reduced the afterdischarge duration of the seizures without significantly modifying the motor convulsions. This effect was noted 24 h after injection and lasted for up to 48 h. Administration of γ-vinyl-GABA in structures 1.5 mm distant from the substantia nigra had no effect on kindled seizures. It is suggested that the substantia nigra may intervene in a negative feedback system that tends to suppress the paroxysmal activity initiated from the amygdala.     

微型膜式氧合器在大鼠体外循环中的应用

目的建立大鼠体外循环（cardiopulmonary bypass，CPB动物模型）。方法选用雄性SD大鼠，经右颈静脉、股静脉插管引流，股动脉插管灌注建立CPB。CPB使用微型膜式氧合器，运转1h，期间监测血流动力学指标、心电图、血气及电解质的变化。结果8只大鼠CPB中血流动力学稳定，血气结果满意，完全符合满意CPB标准，停机后心血管和呼吸功能恢复。结论利用微型动物膜式氧合器建立的大鼠CPB模型，具有安全、稳定、简单、实用的特点，是进行与CPB相关脏器损伤研究可靠的实验平台。     

白藜芦醇调控癫痫持续状态大鼠海马内源性巯醇抗氧化剂(酶)的作用

目的 探讨白藜芦醇(Res)在氯化锂联合重复低剂量匹罗卡品诱导癫痫持续状态大鼠模型中的抗氧化作用及其机制.方法 54只SD大鼠随机分为 3 组:对照组、癫痫模型组和Res治疗组,采用氯化锂联合重复低剂量匹罗卡品腹腔注射制备癫痫持续状态大鼠模型,Res治疗组同时给予白藜芦醇预防性治疗.对大鼠按痫性分级标准进行行为学观测;...     

放射性131Ⅰ诱导的甲状腺功能减退大鼠模型的构建

背景：目前,甲状腺功能减退动物模型造模过程中动物及药物剂量的选择各有差异,造模成功与否的标准也未得到统一。目的：探讨放射性¹³¹Ⅰ诱导大鼠甲状腺功能减退模型的可行性和最佳剂量。方法：健康雄性SD大鼠40只,随机分为5组,每组8只。其中4组大鼠分别腹腔注射4.625,9.25,12.95,18.5 MBq ¹³¹Ⅰ溶液,构建甲状腺功能减退模型;另一组大鼠为正常对照组注射等量生理盐水。分别于注射¹³¹Ⅰ前、注射¹³¹Ⅰ后4,8,12,16周测量大鼠体质量变化,采用ELISA试剂盒法分别测定血清中促甲状腺激素、FT3、FT4、丙氨酸氨基转移酶、门冬氨酸氨基转移酶、碱性磷酸酶、尿素氮、肌酐、尿酸水平,通过甲状腺¹³¹Ⅰ静态显像评价甲状腺功能,苏木精-伊红染色观察甲状腺组织形态学改变。结果与结论：(1)与对照组相比,第8,12,16周9.25 MBq、12.95 MBq、18.5 MBq组大鼠体质量下降(P <0.05);9.25 MBq、12.95 MBq、18.5 MBq组大鼠...     

Developmental characteristics of topographic EEG in school-age children using an autoregressive model

Summary Developmental characteristics of the resting EEG were investigated in 47 school-age children using statistical analysis. Total and component EEG power were statistically analyzed between the subject groups from 7 to 14 year-old using our autoregressive pattern discrimination system. In early school-age children, significant topographic differences were seen in theta waves, while in late school-age children, the differences were found in alpha waves in the frontal and occipital regions, and in beta waves in the frontal region.