实验性癫痫大鼠不同脑区的L-ENK表达及意义

目的 采用戊四氮（PTZ）雳发大鼠癫痫。动态观察了部分脑区亮氨酸-脑啡肽（L-ENK）含量的变化，以探讨L-ENK与癫痫的关系。方法 清洁级近交系雄性SD大鼠50只，分为对照组（A组，10只）及实验组（40只）。实验组按体重50mg／kg腹腔注射PTZ1次，对照组腹腔注射同等容量的生理盐水。根据癫痫发作分级，0～1级7只为B组，立即取脑；2级以上发作33只，随机于癫痫发作后0h（C组，10只），6h（D组，11只），24h（E组，10只）取脑；最后剩下2只（F组）72h取脑。采用放射免疫方法，动态观察脑组织中海马、中脑、纹状体和额叶中L-ENK的改变；S-P免疫组化染色法染色，观察各组大鼠海马CAI区L-ENK的表达。结果 发现应用PTZ后，癫痫大发作大鼠及末出现大发作的大鼠额叶、海马、中脑及纹状体中L-ENK含量均明显升高，两组间除中脑外无明显差异；但大发作大鼠的海马、中脑及纹状体L-ENK含量显著高于对照组。大发作后各脑区的L-ENK含量呈逐渐下降的过程。结论 L-ENK可能参与了癫痫的发生过程。     

尼莫地平对戊四氮慢性点燃癫痫大鼠行为及脑电图的影响

目的:探讨尼莫地平(nimodipine,NIM)对戊四氮(Pentylenetetrazole,PTZ)慢性点燃癫痫大鼠行为及脑电图的影响。方法:64只动物随机分为正常对照组(A组)、PTZ单纯致痫组(B组)、NIM1.25mg/kg干预组(C组)、NIM2.5mg/kg干预组(D组),通过腹腔注射PTZ建立慢性癫痫动物模型,观察大鼠行为、脑电图及海马组织学改变。结果:腹腔注射NIM可以明显抑制大鼠的痫性放电,减小其痫性发作级别,与正常对照组相比重型发作率明显降低(P<0.01),此外,NIM还能减轻海马的损伤程度。结论:NIM在PTZ慢性点燃癫痫大鼠模型中不仅具有抗惊厥作用,还能减轻海马的损伤程度。     

氯喹对戊四氮慢性致痫大鼠脑皮质及海马腺苷激酶表达的影响

目的 观察氯喹对戊四氮致痫大鼠皮质和海马区腺苷激酶(ADK)表达的影响,探讨ADK与癫痫发作的关系及氯喹在癫痫发生过程中的作用. 方法 30只健康雄性SD大鼠按照随机数字表法分为对照组、戊四氮(PTZ)致痫组和氯喹干预组,每组10只.观察大鼠行为学表现,记录其脑电改变,采用免疫组化法检测3组大鼠皮质和海马区ADK的表达. 结果 对照组大鼠无癫痫发作,PTZ致痫组大鼠出现Racine评分中Ⅳ～Ⅴ级严重发作,氯喹干预组大鼠出现Ⅰ～Ⅲ级发作,差异有统计学意义(P＜0.05).PTZ致痫组大鼠脑电记录呈频发高幅的痫样波,氯喹干预组大鼠脑电记录显示慢波、小棘波.PTZ致痫组大鼠脑内ADK表达明显增强,以海马区最为显著,与对照组相比差异均有统计学意义(P＜0.05).氯喹干预组大鼠脑内ADK表达降低,但距离正常水平仍有较大差距,与对照组相比差异有统计学意义(P＜0.05). 结论 癫痫脑组织中存在ADK的表达异常,氯喹可以抑制这种表达,有效控制癫痫发作.     

PTZ致痫大鼠海马区神经元损伤的研究

目的:为了解癫痫(Epilepsg,EP)发生后大脑海马区的损害情况,本文采用戊四氮诱导建立大鼠癫痫模型, 采用酶标免疫吸附法(EIA)和免疫组化染色法检测NSE的改变,使用电子显微镜研究超微结构的改变,探讨EP后脑组织的损坏情况。方法:雄性SD大鼠54只,随机分为对照组(A=13)及实验组(41只)。实验组腹腔注射(IP)PTZ 50mg/ kg体重1次,对照组IP生理盐水。根据EP发作分级,0-1级7只,视为B组,立即取脑:2级以上发作34只,随机抽样于EP发作后0h(C=10);6h(D=14);24h(E=10)断头。取脑前抽取躯干血,分离海马。A、D组随机抽取3只进行电镜观察。采用EIA法测试血清和海马NSE-值;以S-P免疫组化染色法染色,观察各组大鼠海马的NSE表达。结果:实验组海马和血清的NSE均明显高于对照组:即使无EP大发作,但有兴奋症状的大鼠,NSE亦高于对照组,表明也有脑组织损害。免疫组化染色显示:对照组海马组织几乎未见NSE的表达,即大发作后海马的NSE表达明显增加,但随时间推移而降低。电镜观察发现EP后大鼠海马CA1区的神经元细胞出现水肿、基质密度变淡、微丝溶解消失、细胞器减少、毛细血管周围间隙扩大等明显改变。结论:EP发作对大鼠海马区神经细胞造成了明显的损害。     

癫痫动物模型脑中神经肽Y的动态含量变化及意义

目的:探讨匹罗卡品(PILO)诱发的癫痫大鼠模型脑组织中神经肽Y(Neuropeptide Y,NPY)含量的动态变化及意义,进一步明确NPY与癫痫的关系,为抗癫痫治疗,研制抗癫痫药物提供新途径。方法:健康成年雄性SD大鼠120只,随机分为两组:单纯腹腔注射匹罗卡品组(癫痫模型组);单纯腹腔注射生理盐水组(对照组);注射后根据Racine制定的标准判定是否有癫痫发作,并行脑电图检查,观察有无癫痫样波(棘波,尖波,棘慢波,尖慢波)发放。两组大鼠分别于给药后1h,3h,6h,24h,3d,7d,15d,30d,60d将大鼠麻醉,取出脑组织,对脑组织中的NPY含量进行测定。结果:癫痫模型组60只大鼠中,2只死于癫痫持续状态,其余大鼠可观察到边缘发作行为表现,脑电图有典型的癫痫样波发放,对照组无癫痫发作及癫痫样波发放。癫痫模型组脑NPY含量与对照组相比,差异有显著性P<0.05;癫痫模型组急性期(1h-7d)与慢性期(15d-60d)比较差异有显著性P<0.05,对照组差异无显著性;癫痫模型组与对照组脑中的NPY含量在12h,24h,15d,30d,60d.差异有显著性P<0.05或P<0.01,癫痫模型组脑中的NPY含量各组(各时间段)比较有差异有显著性P<0.05,对照组差异无显著性,癫痫模型组大鼠Ⅳ-Ⅴ级发作与Ⅱ-Ⅲ级发作,脑NPY含量比较,差异有显著性P<0.05。结论:1.神经肽Y与癫痫密切相关,癫痫发作后?     

丙戊酸钠对戊四氮致痫大鼠海马神经元凋亡的影响

目的 探讨丙戊酸钠(VAP)对戊四氮(PTZ)致痫大鼠海马神经元凋亡的影响.方法 将48只成年Wistar大鼠随机平均分为正常对照(NC)组、PTZ组和VAP组,PTZ组和VAP组大鼠腹腔注射阈下剂量的PTZ 35mg/(kg·d),直至达到点燃标准.点燃后,VAP组大鼠经腹腔注入VAP15mg/(kg·d),PTZ组大鼠经腹腔注入生理盐水,30min后,再腹腔注射PTZ诱发癫痫发作.应用免疫组化法检测大鼠海马神经元Fas、Caspase-3和Survivin的表达.结果 PTZ组大鼠海马神经元Fas、Caspase-3阳性细胞数和光密度明显高于VAP组和NC组(均P<0.01),Survivin阳性细胞数和光密度明显低于VAP组(P<0.01),但高于NC组(P<0.05);VAP组大鼠海马神经元Fas、Caspase-3阳性细胞数和光密度明显高于NC组(均P<0.05),Survivin阳性细胞数和光密度明显高于NC组(P<0.01).结论 癫痫发作可以导致大鼠海马神经元的凋亡,而丙戊酸钠有对抗癫痫发作导致细胞凋亡的作用,主要通过促进Survivin的表达和抑制Fas和Caspase-3的表达发挥作用.     

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。     

柴胡总皂甙对戊四氮慢性点燃大鼠海马谷氨酸细胞的影响

目的研究柴胡总皂甙对戊四氮(PTZ)慢性点燃癫痫模型大鼠海马区谷氨酸(Glu)阳性细胞表达的影响。方法48只健康SD大鼠被随机分为6组,即空白组(A组)、生理盐水组(B组)、丙戊酸钠(VPA)组(C组)和柴胡总皂甙高、中、低三种剂量组(D组、E组、F组),每组8只,除A组不做处理外,其他各组采用腹腔注射PTZ慢性点燃造模,造模同时给予VPA、柴胡总皂甙等不同处理因素,连续4周后取脑组织切片进行Glu免疫组化染色,从阳性细胞数、灰度值分析结果。结果在CAl区,B组海马阳性细胞数高于A、C、D、E、F组,有显著性差异(P＜0．05),B组海马各区阳性细胞灰度值低于其他各组,与A、C、D各组比较,有显著性差异(P＜0．05)；而在CA2区和DG区,B组阳性细胞数、灰度值与各组差异无统计学意义。结论柴胡总皂甙可以影响PTZ点燃大鼠海马CA1区的Glu表达水平,从而抑制PTZ慢性点燃大鼠的痫性发作。     

白藜芦醇对戊四氮致痫大鼠脑脊液、血清S1OOB蛋白的影响

目的 研究白藜芦醇(Res)对戊四氮致痫大鼠脑脊液、血清S100B蛋白的影响.方法 采用戊四氮(PTZ)腹腔注射建立慢性癫痫模型,造模成功后予以Res(15 mg/kg·d)灌胃干预10 d;采用酶联免疫吸附法测定脑脊液、血清S100B蛋白含量,海马标本行Nissl染色.结果 经28 d连续给药,18只大鼠符合Racine点燃标准,Res干预组大鼠痫性发作潜伏期明显延长,且发作时间明显低于癫痫模型组、二甲基亚砜组(P＜0.05).海马Nissl染色提示Res干预对大鼠海马CA1、CA3区神经元有保护作用(P＜0.05),而对齿状回保护作用不明显(P＞0.05).Res干预组大鼠脑脊液、血清S100B蛋白含量低于癫痫模型组、二甲基亚砜组(P＜0.05).结论 Res降低PTZ致痫大鼠脑脊液、血清S100B蛋白含量,或许减缓癫痫发作脑损伤发挥神经保护作用.     

亚低温对癫痫持续状态大鼠NPY表达影响的实验研究

目的 研究亚低温对癫痫持续状态大鼠神经肽Y(NPY)表达的影响.方法 45只Wistar成年大鼠随机分为常温对照组(A组,n=5),常温SE组(B组,n=20),亚低温SE组(C组,n=20).建立锂-匹罗卡品大鼠癫痫持续状态模型,应用免疫组织化学染色和逆转录PCR方法检测NPY蛋白和基因在癫痫持续状态形成后0h、6h、12h和24h的表达.结果 免疫组织化学染色和RT-PCR显示,癫痫持续状态形成后0h,大脑皮质区和海马区NPY免疫反应活性、mRNA在低水平表达,3组间差异无显著性(P＞0.05);SE后6h,B组和C组NPY蛋白和基因表达水平均上调;12h时,B组、C组NPY蛋白和基因表达水平明显上调;24h时,B组和C组NPY蛋白和基因在高水平表达.但C组表达水平明显较B组高,差异有显著性(P＜0.05),A组NPY表达水平无明显变化.NPY蛋白和mR-NA的表达呈高度正相关(r=0.87,P＜0.05).同时,亚低温组抽搐发作次数和持续时间均较常温组SE组大鼠短少,死亡率下降.结论亚低温可减轻癫痫持续状态,促进癫痫持续状态大鼠的NPY表达,其具体机制仍需深入研究.     

脑室内灌注神经肽Y对戊四氮致痫大鼠海马结构内脑源性神经营养因子表达的影响

目的 :探讨脑室内注入神经肽 Y(Neuropeptide Y,NPY)对戊四氮 (PTZ)致痫大鼠海马结构内脑源性神经营养因子 (Brain- derived Neurotrophic Factor,BDNF)表达的影响。方法 :将 18只健康雄性 Wistar大鼠随机分为 NPY实验组 (n=10 )和对照组 (n=8)。 NPY实验组 ,于 PTZ造模前给予侧脑室注射 NPY(6 nmol/ 10μl) ,对照组给予等容量生理盐水。侧脑室注射后 5分钟 ,腹腔注射 PTZ(6 0 mg/ kg) ,观察痫性发作持续时间并于痫性发作后 2小时处死动物。用免疫组化方法观察海马齿状回 (DG)和 CA1区 BDNF的免疫反应性。结果 :NPY实验组痫性发作时间短于对照组 (P<0 .0 0 1)。实验组 DG颗粒细胞层、分子层的 BDNF免疫反应性高于对照组相应各层 ,各组比较有显著性差异。实验组CA1区锥体细胞层、放射层及腔隙层免疫反应性均高于对照组相应各层 ,各组比较有显著性差异。实验组 DG分子层和 CA1区放射层及腔隙层免疫反应性分别高于颗粒细胞层和锥体细胞层免疫反应性 ,各组比较有显著性差异。 CA1区分子层未见 BDNF免疫反应性表达。结论 :脑室内注射 NPY可缩短痫性发作时间 ,并促进致痫大鼠海马内 BDNF的表达。     

钾通道Kv1.2与致痫大鼠发病相关性的研究

目的通过检测戊四唑(PTZ)致痫大鼠钾通道Kv1.2蛋白表达,探讨钾通道Kv1.2与癫痫发病的相关性。方法 40只SD大鼠分成实验组30只和正常对照组10只。实验组30只大鼠通过腹腔注射PTZ建立全身强直阵挛发作大鼠癫痫模型,取成功致痫鼠24只均分成3组,分别于致痫后3个时间段(1h、24h、48h)取脑组织。用免疫组化法和Western blot法检测大鼠钾通道Kv1.2蛋白。结果实验组大鼠海马区钾通道Kv1.2蛋白表达水平在致痫后3个时间段(1h、24h、48h)均明显低于对照组(P0.05)。实验组大鼠海马区钾通道Kv1.2蛋白表达水平在致痫后3个时间段(1h、24h、48h)之间无显著性差异(P0.05)。结论大鼠海马区钾通道Kv1.2表达的减少与全身强直阵挛发作大鼠癫痫发病密切相关。     

Erdosteine ameliorates PTZ-induced oxidative stress in mice seizure model

The role of oxygen-derived free radicals has been suggested in genesis of epilepsy and in the post seizure neuronal death. The aim of this study was to investigate whether erdosteine has a preventive effect against epilepsy and postepileptic oxidative stress. The mice (n=27) were divided into three groups: (i) PTZ-induced-epilepsy group (n=9); (ii) PTZ-induced-epilepsy+erdosteine group (n=9); (iii) control group (n=9). The animals were observed for a period of 30 min for latency to first seizure onset, total seizure duration, the number of seizure episodes. Then they were sacrificed and the brains were quickly removed, and frozen for biochemical analysis. Malondialdehyde (MDA), nitric oxide (NO), superoxide dismutase (SOD) and xanthine oxidase (XO) activities were carried out in the brain tissue. The latent period between PTZ induction and seizure are longer in the PTZ+erdosteine group than in PTZ-induced-epilepsy group (P<0.05). Biochemical analyses of brain tissue, revealed a significant increase in the MDA, XO and NO levels in the PTZ group according to erdosteine group. SOD level did not change in this group. While MDA and XO levels are significantly lower, SOD level is significantly higher in the PTZ+erdosteine group compared to PTZ and control groups (P<0.01). The present study demonstrated that erdosteine treatment both may increase latent interval between seizures and may decrease oxidative stress, thus may ameliorate neuronal death in brain during seizures. It may be used as an adjunct therapy in epilepsy.     

硫丙咪胺对戊四氮致痈幼鼠海马GFAP、c—fos表达和学习认知的影响

目的：观察硫丙咪胺（Thi）对戊四氮（PTZ）致痫幼大鼠海马胶质原纤维酸性蛋白（GFAP）、c-fos表达及学习认知的影响。方法：发育期SD幼鼠40只随机分为对照组、PTZ致瘸组、TM 30mg·kg^-1干预组和Thi15mg·kg^-1干预组（均n=10）。观察各组大鼠瘸样行为，水迷宫实验观察学习认知能力、免疫组化检测海马GFAP和c-fos的变化。结果：对照组无痫样发作，阿z致痫组有重度发作，Thi30mg·kg^-1干预组有轻度发作（P〈0．05）；水迷宫实验中，PTZ致痫组寻找平台潜伏时间延长和通过平台次数减少，Thi30和15mg·kg^-1干预组寻找平台潜伏时间明显缩短，通过平台次数增加明显，差异有统计学意义（P〈0．05）；PTZ致痫组GFAP和c-fos表达明显强于Thi干预组，差异有统计学意义（P〈0．05），其中Tiff30与15mg·kg^-1干预组相比差异也有统计学意义（P〈o．05）；GFAP和c—fos免疫组化表达与大鼠空间学习记忆能力呈正相关。结论：Thi可能通过抑制PTZ致瘸幼大鼠海马GFAP、c—fos表达，减轻癫痫发作程度，提高其学习认知能力。     

Somatostatin, neuropeptide Y, GABA and cholinergic enzymes in brain of pentylenetetrazol-kindled rats

We studied the effect of pentylenetetrazol (PTZ)-induced kindling (35 mg/kg, i.p., daily) on somatostatin-like immunoreactivity (SOM) with special attention to the duration of changes (rats were sacrificed either 10 days or 4 months after the development of kindling) and to transmitters or modulators related to somatostatin (neuropeptide Y (NPY), GABA, choline acetyltransferase (ChAT), acetylcholinesterase (AchE]. In rats sacrificed 10 days after the last kindled seizure, SOM was elevated in frontal cortex and striatum (p less than 0.01); NPY was elevated in frontal cortex, striatum and hippocampus (p less than 0.05) of kindled or prekindled rats (i.e., rats which were treated daily with PTZ but did not express three consecutive generalized seizures). ChAT activity was slightly decreased (p less than 0.05) in cortex. GABA levels and AchE activity were unchanged in kindled cortex. In rats sacrificed 4 months after the development of kindling none of the parameters analyzed differed from controls. The present study suggests that the cortical and striatal neurons containing SOM/NPY are affected by PTZ-kindling. The cortical cholinergic system is affected to a much smaller extent. The neuropeptide changes are not persistent, as is the lowered seizure threshold, so they are probably not involved in the maintainance of the latter.     

匹罗卡品诱发成年大鼠普遍癫痫与难治性癫痫海马神经发生的变化

背景：神经发生包括细胞增殖、迁移、分化和存活等,是人和多数哺乳动物部分脑区产生新生神经元的过程,主要分布在侧脑室下区和海马齿状回。癫痫可导致海马齿状回的神经发生改变,BrdU是目前公认最理想检测未成熟细胞增殖的标记物之一。 目的：观察匹罗卡品诱发成年大鼠癫痫后神经发生的特点,以及普通癫痫与难治性癫痫神经发生的差异。 设计、时间及地点：随机分组,细胞分子生物学实验,于2006-08/2007-06在华中科技大学同济医学院中心实验室及附属协和医院中心实验室完成。 材料：选用健康Sprague-Dawley雄性大鼠100只,随机分为2组,对照组8只,实验组92只,分为普通癫痫组,自发发作组,难治性癫痫组和非耐药组。匹罗卡品为武汉晶美公司产品,兔抗鼠BrdU抗体为美国sigma公司产品,辣根过氧化物酶标记的羊抗兔IgG为武汉博士德公司产品。 方法：对照组大鼠腹腔注射生理盐水;实验组腹腔注射匹罗卡品15mg/kg,最多注射4次,出现癫痫持续状态的大鼠注射水合氯醛终止发作。致癫大鼠癫痫发作终止后6h腹腔注射BrdU,观察自发发作情况。以脑电图、自发发作频率持续时间,无自发发作为普通癫痫组,有自发发作为自发发作组。行卡马西平灌胃2周并记录发作频率,对卡马西平治疗无效,发作频率减少<50%的为难治性癫痫组,治疗有效即发作频率减少>50%为非耐药组。普通癫痫组分别于注射后第1,2,3,7,14,21和28d取鼠脑海马部做冠状连续切片。非耐药组和难治性癫痫组的大鼠,再次行腹腔注射BrdU,每次为50mg/kg,连续注射4次,每次间隔2h,48h后取脑海马部制作石蜡切片。 主要观察指标：脑海马部切片行免疫组化染色,镜下观察不同时间点BrdU阳性细胞的分布、形态和数量及注射匹罗卡品后大鼠癫痫发作状况。 结果：匹罗卡品第1次注射后所有大鼠无癫痫发作,第2次注射发作16只,第3次注射发作42只,第4次注射后发作11只,14只大鼠4次注射仍无癫痫发作,9只大鼠癫痫持续状态后死亡,77只进入结果分析。神经发生主要位于海马颗粒细胞层和齿状回。与对照组比较,普通癫痫组BrdU阳性细胞明显增多（P<0.01）,难治性癫痫组新生细胞较普通癫痫组明显减少（P<0.01）,神经发生减少。癫痫后第2天BrdU 阳性细胞数目开始增加,14-15d达到高峰,1个月后回到初始水平。 结论：与普通癫痫相比,难治性癫痫可导致神经发生减少。     

Elevated neuropeptide Y and corticotropin-releasing factor in the brain of a novel epileptic mutant rat: Noda epileptic rat

Noda epileptic rat (NER) is a new epileptic rat strain, which was developed by inbreeding rats with spontaneous tonic-clonic seizures in a stock of Crj:Wistar. In the present study, possible changes of two neuropeptides, neuropeptide Y (NPY) and corticotropin-releasing factor (CRF), in the brains of NER were investigated. Increased contents of immunoreactive (IR) NPY were found in the striatum and amygdala of 8-week NERs with partial seizure, while these changes extended to the limbic region including hippocampus in 16-week NERs with fully developed generalized tonic-clonic seizure. IR-CRF were elevated only in the entorhinal and pyriform cortex of both 8-week and 16-week NERs. Generalized tonic-clonic seizure in NERs induced a transient increase of NPY mRNA in the granular layer of dentate gyrus. These results suggest that NPY metabolism in the limbic brain contributes to the seizure susceptibility in this model of epilepsy.     

养血清脑颗粒和丙戊酸联用对戊四氮点燃癫痫大鼠的影响

目的 探讨养血清脑颗粒(YXQNG)联用丙戊酸(VPA)对戊四氮(PTZ)慢性点燃模型大鼠癫痫发作、EEG、认知功能及颞叶、海马T型Ca2+通道蛋白(Cav3.2)表达的影响. 方法 成年雄性SD大鼠40只按随机数字表法分为PTZ组、VPA组、VPA+YXQNG组、NS组,每组10只.前3组大鼠腹腔注射PTZ溶液制作慢性点燃模型,VPA组大鼠在注射PTZ前1 h给予VPA灌胃;VPA+YXQNG组除给予VPA外,注射PTZ前1.5 h给予YXQNG灌胃;NS组腹腔注射生理盐水,每天一次.8周后观察各组大鼠的行为学变化、Y型电迷宫检查大鼠的正确反应率、捕记EEG并应用免疫组化染色检测颞叶和海马Cav3.2的表达. 结果 给药8周后PTZ组大鼠全部达到完全点燃(连续3 d出现Ⅳ级发作或达到Ⅴ级发作),VPA组和VPA+YXQNG组大鼠仅出现0～Ⅱ级发作;Y型电迷宫检查结果显示VPA+YXQNG组大鼠正确反应率高于PTZ组,差异有统计学意义(P＜0.05);EEG结果显示PTZ组大鼠癫痫发作时EEG有明显异常放电,总功率高于用药前,差异有统计学意义(P＜0.05).VPA组、VPA+YXQNG组大鼠用药前后EEG总功率的差值均高于PTZ组,差异有统计学意义(P＜0.05);免疫组化染色结果显示VPA组、VPA+YXQNG组大鼠颞叶和海马Cav3.2表达低于PTZ组,VPA+YXQNG组大鼠颞叶和海马Cav3.2表达低于VPA组,差异均有统计学意义(P＜0.05). 结论 YXQNG和VPA联用能降低癫痫大鼠发作级别、改善认知功能、减少脑部异常放电并降低脑组织Cav3.2水平,有抗癫痫和脑保护作用.

Abstract：

Objective To explore the effet of Yangxue Qingnao granule (YXQNG) on seizures and cognition function of pentylenetetrazole (PTZ)-kindled chronic epileptic rats models, expression of Cav3.2 in the hippocampus and the temporal lobe of these rats, and EEG features of the rats. Methods Forty healthy adult male SD rats were equally divided into 4 groups at random: PTZ group, VPA treatment group, VPA+YXQNG treatment group, normal saline (NS)-control group (n=10). PTZ solution was intraperitoneally injected for 8 weeks to induce the kindling model in the above 3 groups except the NS-control group. VPA by intragastric administration was given to the rats in the VPA treatment group 1 h before PTZ injection; YXQNG and VPA by intragastric administration were given to the rats in the VPA+YXQNG treatment group 1.5 h before PTZ injection. Behavioral changes of the rats were observed 8 weeks after PTZ injection; accuracy rate of response of the rats were examined by electric maze test;EEG was performed; and the expression ofT-type Ca2+ channel protein (Cav3.2) in the temporal lobe and hippocampus was detected by immunohistochemical staining. Results Rats in the PTZ group appeared grade Ⅳ or Ⅴ seizures for 3 consecutive d, and rats in the VPA treatment group, VPA+YXQNG treatment group appeared grade 0-Ⅱ seizures. The accuracy rate of response of the rats in the VPA+YXQNG treatment group was significantly higher than that in the PTZ group (P＜0.05). EEG indicated that paradoxical discharge was noted in rats of PTZ group when seizures appeared, and the total power at the time was obviously higher than that before PTZ injection (P＜0.05). The D-value of total power of EEG in rats of the VPA treatment group and VPA+YXQNG treatment group before and after treatment was significantly higher than that in the PTZ group (P＜0.05). And the level of Cav3.2 in the temporal and hippocampus in rats of the VPA treatment group and VPA+YXQNG treatment group was significantly lower than that in the PTZ group (P＜0.05); as compared with that in the VPA treatment group, the expression of Cav3.2 in the temporal and hippocampus in rats of the VPA+YXQNG treatment group was significantly reduced (P＜0.05). Conclusion The combination use of YXQNG and VPA can decrease the seizure stage, the paradoxical discharge of the brain and the level of Cav3.2 in brain tissue,and improve the cognitive function of the PTZ-kindled rats, indicating that using VAP and YXQNG simultaneously can treat epileptic seizure and protect the neurons.