下丘脑内基因转染对大鼠颞叶癫痫海马病理变化的影响

目的 探讨下丘脑过度兴奋对于颞叶癫痫海马病理变化的影响。方法 利用仙台病毒(HVJ)-脂质体转染法在下丘脑的乳头体内转染兴奋性氨基酸受体亚基GluR2Q，研究其对于海人酸(kainic acid，KA)致痫鼠海马病理变化的影响。结果 GluR2Q能加重KA引起的海马齿状回损伤，但是对CA1及CA3区神经元损伤有一定程度的改善作用。结论 GluR2Q可以使下丘脑兴奋性增强，通过对海马齿状回的抑制解除作用促进癫痫波在海马内的传播。     

癫对雌性大鼠生殖内分泌的影响

目的探讨癫对雌性大鼠生殖内分泌的影响。方法将动情周期规律的成年雌性Wistar大鼠30只随机分为对照组、模型组,每组各15只。模型组大鼠腹腔注射氯化锂-匹罗卡品建立癫模型,对照组大鼠腹腔注射生理盐水,通过阴道脱落细胞涂片判定两组大鼠动情周期的变化,用在四周后在动情间期处死、采血,采用放免法测定血清性激素水平,分离子宫、卵巢组织,检测大鼠体质量、子宫、卵巢重量及子宫、卵巢组织学结构。结果对照组15只大鼠无癫发作,无死亡,动情周期稳定,模型组有3只死亡,其余12只大鼠造模成功,表现为动情周期紊乱,血清性激素中黄体生成素及孕酮水平下降,体质量减轻,子宫和卵巢重量下降,光镜检查子宫腺体数量变少,卵泡变少,与对照组比较差异均有统计学意义（P均〈0.05）。结论癫对雌性Wistar大鼠生殖发育有损害作用。     

颞叶癫大鼠海马区NCX3 mRNA和蛋白的表达变化

目的:动态观察钠-钙交换体(NCX)mRNA和蛋白在氯化锂-匹罗卡品致模型大鼠海马CA1、CA3及齿状回区表达的变化,探讨其在癫发生发展中的作用。方法:用氯化锂-匹罗卡品制备癫动物模型;应用原位杂交和免疫组化技术检测各时间点NCX3mRNA和蛋白的表达。结果:急性期(6～24h)海马各区NCX3mRNA表达均随时间的延长逐渐减少;进入静止期各区表达趋向回升,慢性反复自发发作期(30、60d)各区表达又出现不同程度的两次下调。除致后6h大鼠海马各区的NCX3蛋白表达无明显变化外,NCX3蛋白变化趋势与NCX3mRNA基本一致。结论:NCX3表达下调可能通过增加神经元钙超载,改变海马神经元的兴奋性,促使癫发生。     

选择性杏仁核海马切除术治疗颞叶内侧型癫疗效分析

目的探讨选择性杏仁核海马切除术对治疗颞叶内侧型癫的疗效。方法选择颞叶脑电异常与海马硬化同侧病例10例,其中单纯部分性发作继发全身性发作2例,复杂部分性发作5例（3例继发全身性发作）,全身强直-阵挛性发作3例。经颞底海马旁回入路切除杏仁核海马。结果术后病人均恢复良好。所有病例均随访1a以上,6例发作完全缓解（EngelⅠ、Ⅱ级,60%）,2例缓解明显（EngelⅢ级,20%）,1例轻度缓解（EngelⅣ级,10%）,1例病人术后半年自行停药造成癫复发。结论对伴同侧海马硬化的颞叶内侧型癫患者行选择性杏仁核海马切除术（经颞底海马旁回入路）疗效显著,可改善性发作情况。     

慢性癫大鼠海马电压依赖性钙通道α1亚单位mRNA的研究

目的:研究Ca2+及其通道mRNA在癫发病中的作用。方法:采用半定量原位杂交和Northern杂交技术,对戊四氮慢性致大鼠在致不同时段海马内电压依赖性钙通道(VDCC)α1亚单位mRNA的表达变化进行了研究。结果:实验大鼠在癫形成早期海马不同亚区内α1A、α1D、α1E亚单位mRNA表达水平较对照组明显升高,α1B亚单位mRNA在CA1区和齿状回表达水平下降,而α1C-Ⅰ、α1C-Ⅱ表达无明显变化。充分点燃期仅α1B亚单位mRNA水平在CA3区明显升高。最后一次癫发作后第28天,各亚单位无明显变化。结论:在致过程中,VDCCα1亚单位mRNA有不同程度的变化,其结果可能通过改变神经递质的释放模式而在癫灶形成中起一定作用。     

VEEG监测对鉴别癫及非癫发作的意义

目的探讨发作期视频脑电图（VEEG）在诊断癫性发作及发作类型的临床价值。方法对435例发作性疾病患者的临床资料和发作期VEEG进行回顾性分析。结果 VEEG共监测到临床发作733次,发作期脑电图记录到样放电557次。诊断为癫381例,其中癫合并非癫性发作（NES）24例,占6.3%;单纯NES 54例。癫发作类型以部分性发作为主,癫发作及发作间期样放电多见于NREMⅠ、Ⅱ期。结论 发作期VEEG监测对鉴别癫及非癫发作有重要的鉴别意义。     

捆绑并倒悬复合刺激大鼠海马CA3区神经肽Y和Nestin的变化

【目的】应用改良的连续、恒定复合型应激大鼠模型,观察其血浆肾素活性（PRA）、血管紧张素Ⅱ（AngⅡ）水平和海马CA3区的Nestin、NPY表达。【方法】将雌性成鼠束缚并倒悬应激6h/d,用放射免疫法测定急性应激(3d)后血浆PRA、AngⅡ水平,用免疫组织化学方法观察应激3d、21d后CA3区锥体层细胞的Nestin、NPY表达变化。【结果】①急性应激组血浆PRA和AngⅡ水平比正常对照组显著降低（P<0.01）,尤其是PRA水平的变化最明显。②急、慢性应激组海马CA3区的Nestin、NPY免疫阳性反应产物总面积（SA）和体视学光密度（IOD）值均较对照组低（P<0.01）,急性组的组织损害比慢性组更严重。【结论】本研究成功地构建了具有科学理论依据和实用价值的复合性应激大鼠模型,并利用此模型研究证实应激早期大鼠血浆PRA水平的降低;以及急、慢性应激大鼠的NPY、Nestin和NF200表达变化及其变化规律,结果提示NPY及其水解产物在应激所致海马神经元细胞骨架损害和谷氨酸释放过程中可能产生一定的影响。     

情感障碍对癫患者认知障碍影响的事件相关电位研究

目的应用事件相关电位研究和探讨不同抗癫痫药对癫痫患者认知功能的影响以及情感障碍对癫痫患者认知障碍的影响。方法收集湛江中心人民医院神经内科门诊和住院的癫痫患者320例,正常对照组56例。分别进行韦氏智能测定和情感测定,同时结合抗抑郁剂治疗,并应用事件相关电位P300、N400研究,对比分析癫痫患者认知功能和情感障碍的特征。结果386例癫痫患者中,认知水平低于正常者达76．940。结果显示丙戊酸钠、妥泰、卡马西平单药服药组之间认知水平无差异,但药物联合应用认知功能损害严重。且患者职业、文化程度、病程、发作次数、用药选择以及是否存在脑部继发性疾病等对认知功能影响显著。伴随着认知功能和情感障碍,癫痫患者P300和N400潜伏期明显延长、波幅降低。百忧解治疗后R300和N400潜伏期和波幅明显改善,且颞顶叶脑区的波幅明显升高。结论抗癫痫药可造成癫痫患者认知功能的损害,单药应用较联合用药对癫痫患者认知功能损害轻。抗抑郁治疗可改善癫痫患者的认知功能,认知事件相关电位也明显改善。     

癫痫大鼠海马中神经肽Y表达的动态观察及其作用探讨

目的　探讨癫痫大鼠海马中神经肽 Y( NPY)表达的动态变化及其作用 ,以及外源性 NPY对癫痫的影响。方法　将 SD大鼠随机分为 4组 ,采用匹鲁卡品 ( PILO)癫痫动物模型及外源性 NPY侧脑室注入干预法 ,并以免疫组织化学染色观察 NPY的表达。结果　癫痫模型组 PILO注射后 ,门区、CA3区、CA1 区的 NPY阳性细胞数表达增多 ,1 2 h达高峰 ,以后逐渐下降 ,60 d又达一高峰 ;海马颗粒细胞层有 NPY异位表达 ,3 d时最明显 ;NPY干预组门区 NPY阳性细胞数较其对照组表达少。结论　 ( 1 )癫痫发作后急性期海马 NPY表达增多及颗粒细胞层出现异位表达 ,与癫痫发作有关 ;( 2 )外源性 NPY有抗癫痫作用。     

Anti-epileptic effects of neuropeptide Y gene transfection into the rat brain

Neuropeptide Y gene transfection into normal rat brain tissue can provide gene overexpression,which can attenuate the severity of kainic acid-induced seizures.In this study,a recombinant adeno-associated virus carrying the neuropeptide Y gene was transfected into brain tissue of rats with kainic acid-induced epilepsy through stereotactic methods.Following these transfections,we verified overexpression of the neuropeptide Y gene in the epileptic brain.Electroencephalograms showed that seizure severity was significantly inhibited and seizure latency was significantly prolonged up to 4 weeks after gene transfection.Moreover,quantitative fluorescent PCR and western blot assays revealed that the mRNA and protein expression of the N-methyl-D-aspartate receptor subunits NR1,NR2A,and NR2B was inhibited in the hippocampus of epileptic rats.These findings indicate that neuropeptide Y may inhibit seizures via down-regulation of the functional expression of N-methyl-D-aspartate receptors.     

The anti-epileptic actions of neuropeptide Y in the hippocampus are mediated by Y and not Y receptors

Neuropeptide Y (NPY) potently inhibits glutamate release and seizure activity in rodent hippocampus in vitro and in vivo, but the nature of the receptor(s) mediating this action is controversial. In hippocampal slices from rats and several wild-type mice, a Y2-preferring agonist mimicked, and the Y2-specific antagonist BIIE0246 blocked, the NPY-mediated inhibition both of glutamatergic transmission and of epileptiform discharges in two different slice models of temporal lobe epilepsy, stimulus train-induced bursting (STIB) and 0-Mg2+ bursting. Whereas Y5 receptor-preferring agonists had small but significant effects in vitro, they were blocked by BIIE0246, and a Y5 receptor-specific antagonist did not affect responses to any agonist tested in any preparation. In slices from mice, NPY was without effect on evoked potentials or in either of the two slice seizure models. In vivo, intrahippocampal injections of Y2- or Y5-preferring agonists inhibited seizures caused by intrahippocampal kainate, but again the Y5 agonist effects were insensitive to a Y5 antagonist. Neither Y2- nor Y5-preferring agonists affected kainate seizures in mice. A Y5-specific antagonist did not displace the binding of two different NPY ligands in WT or mice, whereas all NPY binding was eliminated in the mouse. Thus, we show that Y2 receptors alone mediate all the anti-excitatory actions of NPY seen in the hippocampus, whereas our findings do not support a role for Y5 receptors either in vitro or in vivo. The results suggest that agonists targeting the Y2 receptor may be useful anticonvulsants.     

Influence of Ganoderma lucidum spores on the levels of neuropeptide Y and somatostatin in brains of seizure rats*★

BACKGROUND： Previous research has revealed that somatostatin can induce epilepsy, and that the levels of neuropeptide Y may increase and become more active in brain areas with epileptic seizures. OBJECTIVE： To observe the effect of Ganoderma luciclum spore powder on the neuropeptide Y and somatostatin content in the cerebral cortex and hippocampal regions of seizure rats induced by pentylenetetrazol （PTZ）. Furthermore, to verify any effect of Ganoderma lucidum spore powder on inhibition of epileptic seizures. DESIGN, TIME AND SETTING： A randomized group animal study was performed in August 2007 in the School of Basic Medical Sciences, Jiamusi University （Jiamusi, Heilongjiang, China）. MATERIALS： Thirty healthy, male, Wistar rats, aged 12 weeks and weighing 180-220 g, were taken as the experimental animals. PTZ （Sigma Company, United States） was used to induce epilepsy. Ganoderma lucidum spores （Leyss, ex Fr variety） were purchased from Jiamusi City Wild Growing Case of the Ganoderma Lucidum （China）. Rabbit anti-somatostatin antibodies and secondary antibodies were purchased from Wuhan Boster Company （China）. Neuropeptide Y radioimmunoassay kit was purchased from Beijing Furui Biotechnology Company （China）. METHODS： Thirty rats were randomly divided into three groups： a control group, an epilepsy model group and a Ganoderma lucidum spore-treated group. Each group contained 10 animals. Rats in the epilepsy model group were treated with intraperitoneal injections of PTZ and gastric perfusion of physiologic saline. In the Ganoderma lucidum spore-treated group, intraperitoneal injection of PTZ and gastric perfusion of Ganoderma lucidum spore powder were administered. The blank control group was only administered with the physiological saline by intraperitoneal injection and gastric perfusion. MAIN OUTCOME MEASURES： Immunohistochemical staining and radioimmunoassay methods were used to observe the changes of somatostatin and neuropeptide Y content in brain tissue of epileptic r     

Influence of Ganoderma lucidum spores on the levels of neuropeptide Y and somatostatin in brains of seizure rats

BACKGROUND: Previous research has revealed that somatostatin can induce epilepsy, and that the levels of neuropeptide Y may increase and become more active in brain areas with epileptic seizures.OBJECTIVE: To observe the effect of Ganoderma lucidum spore powder on the neuropeptide Y and somatostatin content in the cerebral cortex and hippocampal regions of seizure rats induced by pentylenetetrazol (PTZ). Furthermore, to verify any effect of Ganoderma lucidum spore powder on inhibition of epileptic seizures.DESIGN, TIME AND SETTING: A randomized group animal study was performed in August 2007 in the School of Basic Medical Sciences, Jiamusi University (Jiamusi, Heilongjiang, China).MATERIALS: Thirty healthy, male, Wistar rats, aged 12 weeks and weighing 180-220g, were taken as the experimental animals. PTZ (Sigma Company, United States) was used to induce epilepsy. Ganoderma lucidum spores (Leyss, ex Fr variety) were purchased from Jiamusi City Wild Growing Case of the Ganoderma Lucidum (China). Rabbit anti-somatostatin antibodies and secondary antibodies were purchased from Wuhan Boster Company (China). Neuropeptide Y radioimmunoassay kit was purchased from Beijing Furui Biotechnology Company (China).METHODS: Thirty rats were randomly divided into three groups: a control group, an epilepsy model group and a Ganoderma lucidum spore-treated group. Each group contained 10 animals. Rats in the epilepsy model group were treated with intraperitoneal injections of PTZ and gastric perfusion of physiologic saline, In the Ganoderma lucidum spore-treated group, intraperitoneal injection of PTZ and gastric perfusion of Ganoderma lucidum spore powder were administered. The blank control group was only administered with the physiological saline by intraperitoneal injection and gastric perfusion.MAIN OUTCOME MEASURES: Immunohistochemical staining and radioimmunoassay methods were used to observe the changes of somatostatin and neuropeptide Y content in brain tissue of epileptic rats, as well as the morphology of neurons.RESULTS: All 30 rats were involved in the result analysis, without any loss. The number of somatostatin immunoreacted positive cells in the cerebral cortex and hippocampus was significantly increased in the epilepsy model group compared to the blank control group (P<0.01). The number of somatostatin immunoreacted positive cells in cerebral cortex and hippocampus was significantly decreased in the Ganoderma lucidum spore-treated group compared to the epilepsy model group (P<0.01). The contents of neuropeptide Y in cerebral cortex and hippocampus were significantly increased in the epilepsy model group compared to the blank control group (P<0.01). The contents of neuropeptide Y in the cerebral cortex and hippocampus were significantly decreased in the Ganoderma lucidum spore-treated group compared to the epilepsy model group (P<0.05). The epilepsy seizures in the Ganoderma lucidum spore-treated group were obviously reduced compared to the epilepsy model group.CONCLUSION: Ganoderma lucidum spore powder was able to reduce the somatostatin and neuropeptide Y content in the cerebral cortex and hippocampus effectively, so as to achieve an anti-epileptic function and protect neurons from being damaged.     

腺相关病毒-2介导增强型绿色荧光蛋白基因转染神经干细胞的实验研究

目的探讨血清2型腺相关病毒(rAAV2)介导增强型绿色荧光蛋白(EGFP)基因转染神经干细胞后绿色荧光蛋白(GFP)的表达规律及转染对神经干细胞生物学特性的影响.方法应用荧光显微镜观察转染神经干细胞的绿色荧光蛋白表达情况.传代后第10天,比较转染组和对照组子代细胞形成的神经球数;分化后第14天行免疫组织化学检查,比较转染组和对照组神经元和胶质细胞的比例.结果转染后第3天,可观察到神经球发出绿色荧光,强度逐渐增强,并在第9天达到稳定状态(90%的神经球发出强弱不等的荧光).传代后第10天,转染组子代细胞每井形成的神经球数为(17.83±1.35)个,与对照组比较无显著性差别(P＞0.05).分化后第14天,转染组神经元、胶质细胞的胞体与纤细突起均可见绿色荧光,转染组神经元与胶质细胞比例分别为35.3%±3.1%和55.4%±7.3%,与对照组比较无显著性差别(P＞0.05).结论rAAV2-EGFP可以稳定、高效转染神经干细胞,且不影响神经干细胞的生物学特性,是神经干细胞理想的基因标记方法.     

Combined gene overexpression of neuropeptide Y and its receptor Y5 in the hippocampus suppresses seizures

We recently demonstrated that recombinant adeno-associated viral vector-induced hippocampal overexpression of neuropeptide Y receptor, Y2, exerts a seizure-suppressant effect in kindling and kainate-induced models of epilepsy in rats. Interestingly, additional overexpression of neuropeptide Y in the hippocampus strengthened the seizure-suppressant effect of transgene Y2 receptors. Here we show for the first time that another neuropeptide Y receptor, Y5, can also be overexpressed in the hippocampus. However, unlike Y2 receptor overexpression, transgene Y5 receptors in the hippocampus had no effect on kainate-induced motor seizures in rats. However, combined overexpression of Y5 receptors and neuropeptide Y exerted prominent suppression of seizures. This seizure-suppressant effect of combination gene therapy with Y5 receptors and neuropeptide Y was significantly stronger as compared to neuropeptide Y overexpression alone. These results suggest that overexpression of Y5 receptors in combination with neuropeptide Y could be an alternative approach for more effective suppression of hippocampal seizures.     

Infusion of neuropeptide Y into CA3 region of hippocampus produces antidepressant-like effect via Y1 receptor

A couple of papers indicate that patients with depression show a decrease in serum neuropeptide Y (NPY). To study the role of NPY in depression, we examined the effects of infusion of NPY into the hippocampus of learned helplessness (LH) rats (an animal model of depression). Infusion of NPY into the cerebral ventricle of LH rats showed antidepressant-like effects. Infusion of NPY into the CA3 region, but not the dentate gyrus (DG), produced antidepressant-like effects in the LH paradigm. Infusion of NPY did not affect locomotor activity or aversive learning ability. Coadministration of BIBO3304 (a Y1 receptor antagonist) with NPY to the CA3 region blocked the antidepressant-like effects of NPY, whereas coadministration of NPY with BIIE0246 (a Y2 receptor antagonist) to the CA3 region failed to block antidepressant-like effects. Furthermore, infusions of [Leu(31) Pro(34)]PYY (a Y1 and Y5 receptor agonist) alone and BIIE0246 alone into the CA3 region produced the antidepressant-like effects in LH rats. These results suggest that infusion of NPY into the CA3 region of hippocampus of LH rats produces antidepressant-like activity through Y1 receptors and attenuating effects through Y2 receptors.     

Altered expression of neuropeptide Y,Y1 and Y2 receptors,but not Y5 receptor,within hippocampus and temporal lobe cortex of tremor rats

As an endogenous inhibitor of glutamate-mediated synaptic transmission in mammalian central nervous system, neuropeptide Y (NPY) plays a crucial role in regulating homeostasis of neuron excitability. Loss of balance between excitatory and inhibitory neurotransmission is thought to be a chief mechanism of epileptogenesis. The abnormal expression of NPY and its receptors observed following seizures have been demonstrated to be related to the production of epilepsy. The tremor rat (TRM) is a hereditary epileptic animal model. So far, there is no report concerning whether NPY and its receptors may be involved in TRM pathogenesis. In this study, we focused on the expression of NPY and its three receptor subtypes: Y1R, Y2R and Y5R in the TRM brain. We first found the expression of NPY in TRM hippocampus and temporal lobe cortex was increased compared with control (Wistar) rats. The mRNA and protein expression of Y1R was down-regulated in hippocampus but up-regulated in temporal lobe cortex, whereas Y2R expression was significantly increased in both areas. There was no significant change of Y5R expression in either area. The immunohistochemistry data showed that Y1R, Y2R, Y5R were present throughout CA1, CA3, dentate gyrus (DG) and the entorhinal cortex which is included in the temporal lobe cortex of TRM. In conclusion, our results showed the altered expression of NPY, Y1R and Y2R but not Y5R in hippocampus and temporal lobe cortex of TRM brain. This abnormal expression may be associated with the generation of epileptiform activity and provide a candidate target for treatment of genetic epilepsy.     

Ontogeny of a novel peptide, neuropeptide Y (NPY) in rat brain

Changes in the concentration of a newly discovered peptide, neuropeptide Y (NPY) have been determined in the developing rat brain using a recently developed radioimmunoassay and chromatographic analysis. NPY was present in the brain stem (14.8 ± 5.6 pmol/g) and diencephalon (12.1 ± 12.1 pmol/g) in the earliest embryos studied (14 days postconception), but appeared only on the 19th day postconception in the cerebral cortex. The concentrations of NPY showed a rapid postnatal rise in all regions examined. The finding of NPY early in the development of the embryonic rat brain and particularly in caudal regions has some similarities to the pattern of development of the catecholaminergic system.     

垂体腺瘤中NPY的表达和细胞定位

目的探讨不同病理类型垂体腺瘤中NPY的表达水平及其在瘤细胞中的定位特点。方法收集我科2003年~2005年诊治的垂体腺瘤57例。所有病例均经手术和病理确诊。肿瘤标本进行NPY免疫组化染色。6例垂体瘤标本进行免疫电镜研究。结果本组34/57例垂体腺瘤的NPY呈阳性表达,占59.65%。NPY阳性表达率在促性腺细胞腺瘤中最高达91.7%,生长激素腺瘤和促肾上皮质激素腺瘤次之均为66.7%,零细胞腺瘤和混和性腺瘤均为50%,泌乳素腺瘤表达率最低25%。NPY在不同类型垂体腺瘤中的表达差异有统计学意义(P<0.05)。NPY在亚细胞水平的定位:4/6例(GH腺瘤2例,GH-PRL混合性腺瘤1例,PRL腺瘤2例和促性腺细胞腺瘤1例)NPY免疫电镜结果呈阳性,胶体金颗粒呈散在或小簇状分布,主要存在于胞质中分泌颗粒上,此外,粗面内质网和细胞基质中也偶见散在的胶体金颗粒,其余细胞器很少有非特异吸附的胶体金颗粒。2例PRL腺瘤偶见NPY阳性颗粒。结论①不同类型垂体瘤的NPY表达各有差异。②促性腺细胞腺瘤中的NPY表达水平较高,泌乳素腺瘤中NPY表达较低。