【作者】 曾凡猛；

【导师】 梁军潮；

【作者基本信息】 南方医科大学 ， 神经外科， 2010， 硕士

【摘要】 第一章锂—匹罗卡品诱导实验性颞叶癫痫大鼠模型的建立及致痫大鼠早期死亡原因分析[目的]研究锂—匹罗卡品颞叶癫痫模型致痫后痫性发作的行为学特点、脑电图变化及致痫大鼠早期死亡原因,从而为建立稳定的锂—匹罗卡品颞叶癫痫模型提供方法及降低致痫大鼠早期死亡率。[方法]将Wistar大鼠采用完全随机分组方法分为生理盐水对照组25只、癫痫模型组80只,癫痫模型组大鼠依次给予氯化锂、硫酸阿托品、匹罗卡品腹腔注射诱发癫痫持续状态(status epilepticus, SE),持续1h后注射地西泮终止发作。其后每天观察其自发性痫性发作(spontaneous recurrent seizures, SRS),于造模前清醒状态、SE当时及SE6W分别对大鼠进行EEG描记。并记录致痫大鼠于SE期、静默期、慢性期死亡情况。[结果]锂—匹罗卡品颞叶癫痫模型组大鼠SE诱发成功率92.5%(74/80)。经过10-20天的静默期后,可观察到Ⅰ-Ⅴ级的反复SRS, EEG在急性期表现为持续丛集性高波幅棘波；慢性期可描记到散发棘波或棘慢波。致痫大鼠于SE期、静默期、慢性期死亡率分别为为21.62%(16/74)、4.06%(3/74)、0%。总死亡率为25.68%(19/74),癫痫模型成功率为68.8%(55/80)。[结论]锂-匹罗卡品颞叶癫痫模型致痫成功率高,致痫后总体死亡率不高,且主要集中于SE期,是一种理想的颞叶癫痫动物模型。第二章伽玛刀照射对颞叶癫痫大鼠脑脊液中谷氨酸及γ-氨基丁酸的影响[目的]研究伽玛刀对颞叶癫痫大鼠脑脊液中谷氨酸及γ-氨基丁酸含量的影响,探讨伽玛刀放射外科治疗颞叶癫痫的机理,为临床实践提供理论依据。[方法]将Wistar大鼠分为生理盐水对照组、手术组、非手术组,每组25只。建立锂—匹罗卡品颞叶癫痫模型,手术组大鼠SE后4周给予伽玛刀治疗,照射靶点为左侧海马,周边计量为20Gy,等剂量曲线为80%。分别于GKS前、GKS后1d、GKS后2W、GKS后4W、GKS后8W观测致痫大鼠癫痫发作频率,并分别于GKS后1d、GKS后1W、GKS后2W、GKS后4W、GKS后8W在枕骨大孔处穿刺采集脑脊液,用液相色谱联合质谱方法检测脑脊液中Glu、GABA含量。[结果]行为学观察发现生理盐水对照组大鼠无癫痫发作。致痫大鼠在伽玛刀照射后4W及8W,手术组大鼠癫痫发作次数显著少于非手术组大鼠癫痫发作次数(P=0.000),而伽玛刀照射后1d、2W手术组大鼠癫痫发作次数与非手术组大鼠癫痫发作次数无显著差异(P>0.05)。脑脊液检查结果示伽玛刀照射致痫大鼠1W后手术组大鼠脑脊液中Glu含量显著低于非手术组大鼠脑脊液中Glu含量(P<0.05),且2W后、4W后、8W后之间也存在显著差异(P=0.000),手术组Glu含量(X=709.579nmol/ml)低于非手术组Glu含量(X=828.620nmol/ml),且手术组大鼠脑脊液中Glu含量随时间延长而减少。照射1W后手术组GABA含量显著高于非手术组大鼠含量出现明显差异(P<0.05),且2W后、4W后、8W后之间也存在显著差异(P=0.000),手术组GABA含量(X=221.866nmol/ml)高于非手术组GABA含量(X=159.730nmol/ml),且手术组大鼠脑脊液中GABA含量随时间延长而增加。[结论]伽玛刀照射致痫大鼠后致痫大鼠脑脊液中Glu含量降低、GABA含量升高先于致痫大鼠癫痫发作频率降低,致痫大鼠中Glu含量降低、GABA含量升高,导致兴奋性神经递质与抑制性神经递质失衡,从而降低致痫大鼠癫痫发作频率。致兴奋性神经递质与抑制性神经递质失衡可能是伽玛刀治疗颞叶癫痫的重要机制之一。更多还原

【Abstract】 Establish the lithium-pilocarpine-induced experimental rat model of temporal lobe epilepsy and analysis the early death of induced epileptic ratsOBJECTIVEStudy of the behavioral seizure characteristics, EEG changes and epileptic causes of early death in rats that were lithium-pilocarpine model of temporal lobe epilepsy after epileptic seizures, So as to provide a way and to reduce early mortality of epileptic rats for establishing a stable lithium-pilocarpine model of temporal lobe epilepsy.METHODSThe Wistar rats were randomly divided into saline control group which contains 25 rats, and epilepsy model group which contains 80, Epilepsy model rats were given lithium chloride, atropine sulfate, pilocarpine by intraperitoneal injection to induce status epilepticus(SE), and to terminate seizures by diazepam 1 hour after the injection of continuous. To observe the spontaneous recurrent seizures(SRS) per day thereafter, and record the EEG trace of the rats at awaking before making model, being status epilepticus, six weeks later. So as to record the deaths of epileptic rats in the SE period, quiet period, and chronic phase.RESULTSThe success rate of SE-induced was 92.5%(74/80) in lithium-pilocarpine model of temporal lobe epilepsy rats. After 10-20 days of the silent period, we can observe I-V grade repeated SRS, EEG in the acute phase showed sustained high volatility spikes clustered; and we can record the dissemination of spikes or spike and slow wave in chronic phase. The mortality of induced epileptic rats was 21.62% in SE period,4.06% in silent period,0% in chronic phase. Total mortality rate was 25.68% (19/74). The success rate of epilepsy model was 68.8%(55/80)CONCLUSIONLithium-pilocarpine model of temporal lobe epilepsy epileptic high success rate, After epilepsy induced, the overall mortality rate of epileptic is not high, and is mainly focused on the SE period, is an ideal animal model of temporal lobe epilepsy.Chapter TwoThe effection of glutamate and y-aminobutyric acid in cerebrospinal fluid of rats with temporal lobe epilepsy by low-dose gamma knife irradiationObjectiveResearch the effection of glutamate and y-aminobutyric acid in cerebrospinal fluid of rats with temporal lobe epilepsy by gamma knife, To investigate the mechanism of Gamma Knife radiosurgery treatment of temporal lobe epilepsy, To provide a theoretical basis for clinical practice.MethodThe Wistar rats were divided into saline control group, surgery group, non-surgical group, each 25 rats, To establish a temporal lobe epilepsy model with lithium-pilocarpine, SE-operated rats were given Gamma Knife after 4 weeks, Irradiated target was the left hippocampus, Out Measurement was20Gy, Isodose curve was 80%, To observe the frequency of epileptic seizures in rats at GKS before, GKS after Id, GKS after 2W, GKS after 4W, GKS post-8W respectively, then puncture and collect the CSF at the foramen at GKS after 1d, GKS after 1W, GKS after 2W, GKS after 4W, GKS post-8W respectively, Detect the Glu,GABA content in CSF by joint mass spectrometry Liquid Chromatography.ResultsBehavioral obseevation found the saline control rats without epileptic seizures. After gamma knife irradiation 4W,the number of epileptic seizures in rats of operation group were significantly different from non-surgical group(P=0.000),The seizure frequency in rats with Gamma Knife irradiation 1d,2W later were no significant difference from non-surgical rats(P> 0.05), The cerebrospinal fluid examination results showed that the Glu content in cerebrospinal fluid of operated rats were significant differences from non-operated group after Gamma irradiation induced epileptic rats 1W(P<0.05), and there were difference 2W later,4W later,8W later. Operation group (Glu content of number 709.579nmol/ml) lower than that of non-operation group (Glu content of number 828.620nmol/ml). And the Glu content in cerebrospinal fluid of the surgical rats deceased with time. The GABA levels of the surgical group with 1W irradiation were significant different from non-operated rats(P <0.05), and there was a significant difference at 2W later,4W later,8W later(P= 0.000), Operation group (GABA content of number 221.866nmol/ml) were higher than non-operation group (GABA content of number 159.730nmol/ml)), CSF GABA levels of the operation group increased with time.ConclusionAfter gamma knife irradiation induced epileptic rats, the Glu content in cerebrospinal fluid of epileptic rats decreased, GABA content increased, that prior to the reduced frequency of epileptic seizures in rats. Glu content decreased, GABA content increased in epileptic rats, cause excitatory neurotransmitter and inhibitory neurotransmitter imbalances, Thereby the frequency of epileptic seizures in rats were reduced. The imbalances between excitatory neurotransmitter and inhibitory neurotransmitter may be one of an important mechanism for temporal lobe epilepsy by gamma Knife treatment.更多还原