低剂量伽玛刀照射对致大鼠皮质及海马神经元c-fos和nNOS表达的影响

目的 通过观察低剂量伽玛刀照射对致痫大鼠大脑皮质及海马神经元c—fos和脑型一氧化氮合酶（nNOS）表达的影响，探讨伽玛刀治疗癫痫的作用机制。方法将44只青霉素致痫大鼠模型等分为实验组和实验对照组大鼠各22只，另取4只正常大鼠作为正常对照组。实验组行伽玛刀照射（周边剂量12Gy）后，应用免疫组化方法，观察大脑皮质及海马神经元c-fos和nNOS表达的变化。结果无论是皮质还是海马，c—fos和nNOS在实验组与实验对照组动物之间，表达均有明显的差别，实验组表达明显少于实验对照组，而后者呈现双高峰现象。结论c—fos和nNOS在伽玛刀治疗癫痫的机制中发挥了重要作用。     

低剂量伽玛刀对致痫大鼠皮层及海马神经元Fos表达的影响

目的通过观察低剂量伽玛刀照射对致痫大鼠大脑皮质及海马神经元cfos表达的影响,初步探讨伽玛刀治疗癫痫的作用机制,为临床治疗提供理论依据。方法以青霉素致痫模型为对象,应用免疫组化方法,检测低剂量伽玛刀照射(周边剂量12Gy)后,大脑皮质及海马神经元cfos表达的变化。实验组和对照组大鼠各22只,正常对照组大鼠4只。结果无论是皮层还是海马,伽玛刀照射后cfos在实验组动物和对照组动物中,表达均有明显的差别,并随时间呈现一定的规律性。结论cfos在伽玛刀治疗癫痫的机理中发挥重要作用。     

低剂量伽玛刀照射对致(癎)大鼠皮质及海马神经元c-fos和nNOS表达的影响

目的 通过观察低剂量伽玛刀照射对致(癎)大鼠大脑皮质及海马神经元c-fos和脑型一氧化氮合酶(nNOS)表达的影响,探讨伽玛刀治疗癫(癎)的作用机制.方法 将44只青霉素致(癎)大鼠模型等分为实验组和实验对照组大鼠各22只,另取4只正常大鼠作为正常对照组.实验组行伽玛刀照射(周边剂量12 Gy)后,应用免疫组化方法,观察大脑皮质及海马神经元c-fos和nNOS表达的变化.结果 无论是皮质还是海马,c-fos和nNOS在实验组与实验对照组动物之间,表达均有明显的差别,实验组表达明显少于实验对照组,而后者呈现双高峰现象.结论 c-fos和nNOS在伽玛刀治疗癫(癎)的机制中发挥了重要作用.     

低剂量伽玛刀照射对癫痫大鼠皮层及海马NMDA受体亚基表达的影响

目的观察低剂量伽玛刀照射对癫痫大鼠皮层和海马N-甲基-D-天氡氨酸(N-methyl-Daspartate,NMDA)受体亚基表达的影响。方法根据动物是否致痫及接受伽玛刀照射,将大鼠分为4组:对照组、伽玛刀组、药物致痫组、伽玛刀+药物组。腹腔连续注射戊四氮(pentylenetetrazole,PTZ)制备癫痫大鼠模型,以双侧额叶为照射靶区对大鼠进行低剂量伽玛刀照射,边缘剂量为15Gy。观察并记录各组大鼠伽玛刀照射前、后癫痫发作情况,并于伽玛刀照射后12周后留取脑组织,分别利用免疫组化及免疫蛋白印迹法对大鼠皮层及海马NMDA受体亚基NR1、NR2A和NR2B进行检测。结果对照组及伽玛刀组大鼠无痫性发作表现,与药物致痫组大鼠相比,伽玛刀+药物组大鼠经低剂量伽玛刀照射后12周,痫性发作明显减轻(P0.05)。与对照组相比,药物致痫组大鼠额叶皮层及海马CA1、CA3区NR1、NR2A和NR2B表达均明显增强(P0.05),阳性神经元数目及平均吸光度值均明显增加(P0.05);与药物致痫组比较,伽玛刀+药物组额叶皮层及海马CA1、CA3区NR1、NR2A和NR2B表达均明显降低(P0.05),阳性神经元数目及平均吸光度值明显减少(P0.05);伽玛刀组与对照组无明显差别(P0.05)。结论癫痫大鼠额叶皮层及海马NR1、NR2A及NR2B亚单位蛋白表达增强,低剂量伽玛刀照射可能引起癫痫大鼠皮层及海马NMDA受体亚基表达减少,这可能是低剂量伽玛刀抑制癫痫发作的分子机制之一。     

伽玛刀对癫痫大鼠模型的治疗作用及机制研究

目的 研究伽玛刀照射对癫痫大鼠的放射生物学作用,探讨伽玛刀治疗癫痫的作用机制.方法 制备海人酸大鼠癫痫模型,利用自行设计的伽玛刀动物定向头架,分别应用100Gy和20Gy的伽玛射线对癫痫大鼠海马组织进行照射,观察大鼠行为学、脑电图、MRI及超微结构、脑组织氨基酸含量及GABA能神经元表达变化.结果 伽玛刀照射后大鼠癫痫发作次数明显减少.100Gy组照射后2个月MRI表现为靶区高信号,20Gy组5个月MRI未见改变.伽玛刀照射后兴奋性氨基酸含量显著低于癫痫组,GABA能神经元表达低于正常,但高于癫痫组.结论 伽玛刀照射对癫痫大鼠具有治疗作用,高剂量伽玛射线(100Gy)对致痫灶有毁损作用.低剂量伽玛射线(20Gy)通过抑制兴奋性神经元释放兴奋性氨基酸使癫痫发作减少.     

中枢组胺对戊四氮点燃大鼠海马GABA能神经元的影响

目的:探讨中枢组胺抗癫痫的作用机制。方法:应用免疫组织化学方法研究中枢组胺对戊四氮(PTZ)致癫痫大鼠海马神经元GABA、GAD67表达的影响。结果:戊四氮致痫组大鼠海马神经元GABA、GAD67的表达量明显低于正常对照组,L-组胺酸干预组明显高于戊四氮致痫组,L-组胺酸干预组与正常对照组之间差异无显著性意义。结论:中枢组胺通过激活海马GABA能神经元来抑制癫痫的发生和发展。     

海马神经元激活物致疒间大鼠大脑皮质和海马γ-氨基丁酸受体的表达

目的 观察大鼠海马神经元在激活物作用下γ-氨基丁酸（GABA。）受体的表达,进一步探讨癫痫发病机制。方法 将大鼠海马神经尢被戊四氮（PTZ）作用后的激活物（实验组）及无血清培养基（对照组）注入大鼠侧脑室后观察其行为、脑电图（EEG）及脑组织GABA,受体表达的变化。结果 实验组大鼠注射后15～30min出现Ⅱ～Ⅲ级惊厥反应,EEG呈短程中高幅尖波、尖慢复合波;对照组的行为及EEG未见异常;各时点实验组大鼠脑组织GABA、免疫反应阳性细胞表达明显低于对照组（均P〈0．05）,对照组GABA,免疫反应阳性细胞广泛分布于大脑皮质、海马回、齿状回。结论 海马神经元激活物具有明显致痫作用,其致痫效应与GABAA受体含量下降有关。     

细胞因子及免疫抑制剂对大鼠脑内神经递质的影响

目的研究白细胞介素-1β(Interleukin-1β,IL-1β)、白细胞介素-6(Interleukin-6,IL-6)、戊四氮致痫和免疫抑制剂抗痫效应过程中谷氨酸(glutamate,Glu)和γ-氨基丁酸(γ-aminobutyric acid,GABA)在大脑皮质及海马内表达的变化,探讨IL-1β、IL-6在癫痫发病中的作用机制及免疫抑制剂的抗痫效应。方法将实验大鼠随机分为6组;即:对照组;IL-1β组;IL-6组;戊四氮组;IL-1ra(IL-1受体拮抗剂) 戊四氮组;地塞米松 戊四氮组。行侧脑室注射相应试剂120min后观察大鼠行为变化,并采用免疫组织化学方法检测大脑皮质及海马内Glu及GABA的表达变化。结果动物行为学观察,IL-1β组、IL-6组癫痫发作程度达中度;戊四氮组癫痫发作程度达重度。IL-1ra 戊四氮组癫痫发作较戊四氮组减轻,地塞米松 戊四氮组无明显癫痫发作。免疫组化染色显示,IL-1β组、IL-6组、戊四氮组Glu表达在大脑皮质及海马较对照组明显升高,GABA表达较对照组明显降低,差异具有显著性意义。IL-1ra 戊四氮组及地塞米松 戊四氮组与单独注射戊四氮组比较,Glu免疫染色减弱,GABA免疫染色增强,有显著性差异。结论IL-1β或IL-6可能通过升高Glu含量并降低GABA的含量参与致痫过程,从而使神经元兴奋性升高促进癫痫发作。免疫抑制剂具有抗痫效应。     

脑室内注射乙酰胆碱受体抗体IgG对大鼠神经元凋亡及一氧化氮合酶表达的影响

目的研究乙酰胆碱受体抗体(AchRab)对大鼠脑内神经元的损害及一氧化氮合酶(NOS)在损害中所起的作用，探讨重症肌无力(MG)中枢神经系统损害的机制。方法将AchRab IgG或健康人的IgG注入大鼠侧脑室。HE染色、TUNEL法检测细胞凋亡；免疫组化方法观察大鼠皮质、海马及杏仁核神经元型一氧化氮合酶(nNOS)和诱导型一氧化氮合酶(iNOS)表达变化。结果2周后实验组皮质、海马及杏仁核凋亡细胞明显增多，对照组仅见少量凋亡。实验组皮质、海马及杏仁核nNOS神经元数目明显减少。实验组及对照组脑内细胞均来见iNOS表达。结论AchRab脑内注射可诱导神经元凋亡；损伤皮质。海马及杏仁核nNOS神经元；但未能诱导脑内细胞iNOS表达。神经元凋亡损害参与了AchRab对中枢神经损害的机制；nNOS神经元的减少，可能与MG认知功能障碍有密切关系；而神经元的损伤可能与NO的毒性作用无关。     

三七皂苷对大鼠缺血脑组织一氧化氮合酶表达的影响——免疫细胞化学与图像分析系统在免疫药理学研究中的应用

应用免疫细胞化学技术与图像分析系统研究三七皂苷的免疫药理学机制。正常组大鼠;实验组大鼠(左颈总动脉结扎并于术前4小时肌注三七皂苷,7mg/kg~(-1))与对照组大鼠(左颈总动脉结扎用等量生理盐水取代三七皂苷)各分4小时组、12小时组、24小时组。各组鼠大脑皮质、海马与尾壳核内神经元性一氧化氧合酶(nNOS)的含量用LSAB法与计算机数字图像分析系统检测。结果显示对照组大鼠海马、尾壳核与大脑皮质内nNOS含量显著下降,三七皂苷可恢复缺血脑组织的nNOS表达,并对缺血脑组织发挥保护作用。同时,本实验还提示免疫细胞化学技术与图像分析系统用于免疫药理学研究是可行的。     

低剂量伽玛刀照射癫癎大鼠脑神经元的超微结构研究

目的探索伽玛刀治疗原发性癫的细胞学机理。方法建立大鼠皮质青霉素局灶性癫癎模型,将SD大鼠随机分为实验组、实验对照组和对照组。照射周边剂量12 Gy, 等剂量曲线为50%。分别于0.5 h~2个月后取靶区皮质及海马标本制备光镜、常规透射电镜样品。结果癫癎模型鼠可见较多凋亡神经元,而癫癎模型鼠经低剂量伽玛刀照射后同时程的神经元改变较轻微,凋亡细胞少见。结论凋亡参与了青霉素致癫癎发作后海马神经元的死亡过程,低剂量伽玛刀照射对抑制神经元的死亡过程有重要作用。     

伽玛刀低剂量照射对致疒间大鼠海马中氨基酸递质的影响

目的观察伽玛刀低剂量照射对致疒间大鼠海马中谷氨酸(Glu)、γ-氨基丁酸(GABA)的影响,探讨伽玛刀治疗癫疒间的作用机制。方法将50只锂-匹罗卡品致疒间大鼠随机分为照射组和非照射组,各25只,另取正常大鼠25只(腹腔注射生理盐水)作为对照组。照射组大鼠进行伽玛刀低剂量照射(周边剂量20Gy),对照组和非照射组大鼠不进行伽玛刀照射。用高效液相色谱-质谱-质谱(HPLC-MS-MS)分析法检测各组大鼠海马Glu、GABA的含量。结果伽玛刀照射2周后,照射组和非照射组大鼠海马中Glu含量均高于对照组,照射组GLu含量低于非照射组,差异具有统计学意义(P0.05);照射组和非照射组GABA含量均低于对照组,照射组GABA含量高于非照射组,差异具有统计学意义(P0.05)。结论伽玛刀低剂量照射通过调节海马中Glu与GABA的平衡,以达到抗癫疒间的作用。     

柴胡疏肝汤对戊四氮致痫大鼠海马及额叶皮质c-fos表达的影响

目的 观察柴胡疏肝汤对戊四氮致痫大鼠海马及额叶皮质c-fos表达的影响.方法 选取经戊四氮诱导制作的癫痫模型大鼠48只,按随机数字表法分成6组:癫痫模型组、德巴金组、定痫丸组、柴胡疏肝汤低剂量组、柴胡疏肝汤中剂量组和柴胡疏肝汤高剂量组,每组各8只;另设正常对照组8只.正常对照组和癫痫模型组常规饲养,其他各组给予药物德巴金,定痫丸,低、中、高剂量柴胡疏肝汤处理,均连续灌胃治疗5周.免疫组化染色检测各组大鼠海马及额叶皮质c-fos的表达情况.结果 戊四氮致痫后大鼠海马及额叶皮质c-fos表达明显增强,而应用中、高剂量的柴胡疏肝汤,德巴金和定痫丸治疗后,大鼠海马及额叶皮质c-fos表达均明显减弱,与癫痫模型组比较差异均有统计学意义(P<0.05),而柴胡疏肝汤低剂量组大鼠海马及额叶皮质c-fos表达无明显减弱.结论 柴胡疏肝汤的抗癫痫作用机制可能与其含有柴胡皂甙及其他多种有效的抗癫痫成份多靶点干预海马及额叶皮质c-fos表达水平有关.

Abstract：

Objective To observe the effect of chaihushugantang on the expression of c-fos in the hippocampus and frontal lobe cortex of pentylenetetrazole (PTZ)-induced epileptic rats. Methods Forty-eight PTZ-induced epileptic rats were randomly divided into 6 groups: epileptic model group,Valproate treatment group, Dingxianwan treatment group, and lower-dose, medium-dose and high-dose chaihushugantang treatment groups (n=8). Normal control group was also employed (n=8). The epileptic rats in the normal control group and epileptic model group were fed normally. Rats of the treatment groups were performed intragastric administration of medicines (Valproate, Dingxianwan and chaihushugantang) for 5 weeks in succession respectively. The expression of c-fos in the hippocampus and frontal lobe cortex of all the rats was observed. Results The expression of c-fos in the hippocampus and frontal lobe cortex of rats in the epileptic model group was significantly increased, while the c-fos expression in the hippocampus and the frontal lobe cortex of rats in the medium-dose and high-dose chaihushugantang treatment groups and Valproate treatment group and Dingxianwan treatment group was significantly decreased as compared with that in epileptic model group (P<0.05). But the c-fos expression in the hippocampus and the frontal lobe cortex of rats in the low-dose chaihushugantang treatment group showed no obvious decrease. Conclusion Chaihushugantang has good antiepileptic effect, might through affecting the c-fos expression in the epileptic rats. The antiepileptic mechanism of chaihushugantang can be related to saikosaponins and other antiepileptic constituents in it.     

Pinealectomy stimulates and exogenous melatonin inhibits harmful effects of epileptiform activity during pregnancy in the hippocampus of newborn rats: an immunohistochemical study

Objectives Epilepsy during the pregnancy is an important problem in clinical practice for newborn individuals. Recently, it has been demonstrated that mothers’ epileptic seizures have some harmful effects on newborns, but present data concerning the effects of epileptic phenomena in pregnant mothers on newborn pups are still limited. The current study was undertaken to investigate the morphological changes in the hippocampus of newborn pups of pinealectomized rats subjected to experimental epilepsy during pregnancy.Methods In this study, rats were randomly divided into four groups (ten animals each): intact control group, epilepsy control group, surgical pinealectomy + epilepsy group, and group with melatonin treatment following pinealectomy procedure. The animals in surgical pinealectomy + epilepsy and melatonin treatment groups underwent a surgical intervention consisting of pineal gland removal. At 1 month after surgical pinealectomy, an acute grand mal epileptic seizure was induced by 400 IU penicillin G administration into their hippocampal CA3 region on the 13th day of their pregnancy in all animals except the intact control animals. On the first neonatal day, the hippocampi were removed and processed for microscopic examination. Nestin expression was analysed in the developing hippocampal tissue.Results Normal migration and hippocampal maturation were determined in the postnatal rat hippocampus in intact control group, but the morphological structure of the hippocampus in the epilepsy control group corresponded to the early embryonal period. It was found that experimental epilepsy and pinealectomy enhanced nestin immunoreactivity, whereas exogenous melatonin treatment (30 μg/100 g body weight, intraperitoneal) inhibited pinealectomy-stimulated nestin expression in CA1 region of the hippocampus.Conclusion These findings suggest that epileptic seizures during pregnancy may cause an impaired hippocampal neurogenesis and neuronal maturation in the newborn, and the negative effects in the postnatal rat hippocampus are more dramatic after pinealectomy of the mother; conversely, melatonin administration suppresses these negative changes. This is the first report investigating the effects of maternal epilepsy during pregnancy in pinealectomized rats on nestin immunoexpression in the newborn rat hippocampus.Presented in part at the 4th Asian-Pacific International Congress of Anatomists (APICA), Kuşadası, Turkey, 7-10 September 2005.     

去松果体对大鼠学习能力及大脑皮质NOS表达的影响

目的探讨去松果体后其功能减退致褪黑素（ＭＴ）分泌减少对大鼠学习能力及大脑皮质一氧化氮合酶（ＮＯＳ）表达的影响。方法将大鼠进行Ｙ型迷宫测试,淘汰学习障碍的大鼠,将学习正常的大鼠随机分二组,实验组手术摘除松果体,对照组给予假手术,饲养４０天后再行Ｙ型迷宫测试,ＳＡＢＣ法检测神经元型一氧化氮合酶（ｎＮＯＳ）表达。结果实验组大鼠学习能力明显低于对照组大鼠（Ｐ〈０．０１）,而大脑皮质、内侧隔核－斜角带核（Ｓ     

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.     

普瑞巴林对慢性颞叶癫癎大鼠海马凋亡调控基因的影响

目的通过观察普瑞巴林对匹罗卡品慢性癫癎大鼠海马区Bcl-2和Bax表达的影响,探讨普瑞巴林治疗癫癎的药理学机制及对大鼠海马神经元的抗凋亡作用。方法采用氯化锂-匹罗卡品化学诱导方法建立慢性颞叶癫癎模型。经腹腔注射普瑞巴林40mg(/kg·d)连续治疗3周,免疫组织化学染色和Western blotting法检测不同处理组大鼠海马区Bcl-2和Bax表达变化。结果与生理盐水对照组比较,模型组大鼠海马区Bcl-2和Bax表达水平显著升高(均P=0.000);与模型组比较,普瑞巴林治疗组大鼠海马区Bcl-2表达水平升高、Bax表达水平降低,组间差异具有统计学意义(均P=0.000)。结论新型抗癫癎药物普瑞巴林可通过降低慢性颞叶癫癎大鼠海马区Bax表达、上调Bcl-2表达而抑制细胞凋亡,发挥神经元保护作用。     

海人酸致大鼠海马结构GABA_B受体亚单位表达的研究

目的:探讨海人酸诱导大鼠颞叶癫(EP)发作后2种γ-氨基丁酸(GABA)受体亚单位GABABR亚单位1a(GBR1a)和GABABR亚单位2(GBR2)在EP发生、发展中的作用。方法:运用原位杂交及免疫组化法,检测EP发作后GABABR亚单位mRNA及蛋白在海马的表达。结果:致早期CA1和CA3区2种亚单位mRNA表达持续低下后逐渐增加,DG区则暂时性下降后很快回升;而免疫反应早期却未见明显改变,随后CA1和CA3区表达处于低水平,DG区和颞叶皮质表达下降后很快恢复。结论:致后2种GABAB受体亚单位基因和蛋白表达上调为颞叶EP的内源性自我保护机制。