丙戊酸钠对戊四氮致癎大鼠海马Bax和Bcl-2表达的影响

目的探讨丙戊酸钠(VAP)对戊四氮(PTZ)致癎大鼠海马Bax和Bcl-2表达的影响.方法将24只成年Wistar大鼠随机分为正常对照(NC)组、PTZ组和VAP组,PTZ组和VAP组大鼠腹腔注射阈下剂量的PTZ 35 mg/(kg·d),直至达到点燃标准.点燃后,VAP组大鼠经腹腔注入VAP 15 mg/(kg·d),PTZ组大鼠经腹腔注入生理盐水,30 min后,再腹腔注射PTZ诱发癫癎发作.应用免疫组化法检测大鼠海马神经元Bax和Bcl-2蛋白的表达.结果大鼠海马神经元Bax阳性细胞数和光密度,PTZ组均明显高于VAP组和NC组(均P<0.01),VAP组明显高于NC组(P<0.05);大鼠海马神经元Bcl-2阳性细胞数和光密度,PTZ组明显高于NC组(P<0.05),VAP组均明显高于PTZ组和NC组(均P<0.01).结论 VAP可以增强癫癎大鼠海马神经元Bcl-2的表达和降低Bax的表达,有对抗癫癎发作导致细胞凋亡的作用.     

锂-毛果芸香碱致痫大鼠早期大脑NG2和O4表达及意义

目的探讨氯化锂-毛果芸香碱(匹罗卡品)致疒0 间大鼠早期大脑少突胶质前体细胞变化及意义.方法对雄性成年SD大鼠先后腹腔注射氯化锂、毛果芸香碱,制成癫癎持续状态动物模型;用免疫荧光组织化学法检测癎性发作后早期大鼠大脑皮质和海马CA1区NG2和O4阳性细胞数量.结果和对照组相比,除癫癎后1 d组外其余各组大鼠脑皮质内NG2和O4阳性细胞都有明显的增加;癫癎后1 d组海马CA1区的阳性细胞数明显减少;癫癎后7 d组皮质和海马CA1区NG2和O4阳性细胞数最多.结论氯化锂-毛果芸香碱致癎大鼠早期大脑NG2和O4表达增加,少突胶质前体细胞增多,并且和观测时间相关.     

电刺激丘脑底核对大鼠杏仁核电点燃的抑制作用

目的 在大鼠杏仁核电点燃癫痫模型中研究丘脑底核高频深部电刺激对点燃的抑制作用及其对Glu、GABA浓度的影响.方法 建立大鼠杏仁核电刺激点燃模型,观察丘脑底核高频深部电刺激对点燃发作的抑制作用,应用高效液相色谱（HPLC）测定大鼠脑组织中Glu、GABA的浓度.结果 对丘脑底核高频深部电刺激（130 Hz,0.2 ms,5 V）能够有效抑制大鼠杏仁核电点燃（P＜0.05）;并可以降低纹状体内Glu浓度,升高纹状体内GABA浓度（P＜0.05）.结论 丘脑底核高频深部脑电刺激能有效抑制大鼠杏仁核电点燃,其作用机制可能与改变Glu、GABA浓度平衡有关.     

托吡酯对癫痫大鼠海马细胞外液氨基酸和神经元凋亡的影响

目的研究托吡酯对癫癎大鼠海马区细胞外液氨基酸和神经元凋亡的影响.方法采用戊四氮(PTZ)致癎模型,大鼠癫癎发作后连续给予托吡酯(TPM)80 mg·kg-1·d-1和卡马西平40 mg·kg-1·d-1,共14 d.以TUNEL方法标记DNA片段,原位检测海马凋亡的神经细胞.脑内微透析技术采集大鼠海马细胞外液,反相高效液相色谱技术测定氨基酸类神经递质的含量.结果TPM组、卡马西平组与对照组比较,凋亡细胞数存在显著差异(P＜0.001),TPM组与卡马西平组相比无显著差异(P＞0.05).TPM可明显升高海马细胞外液γ-氨基丁酸(GABA)水平,并降低谷氨酸(Glu)浓度.结论TPM可减轻大鼠癫癎发作后的神经元损伤,这种作用可能是氨基酸变化的结果;但在我们的实验中,没有发现TPM对癫癎后脑损伤比卡马西平有更明显的神经保护作用.     

生酮饮食对海人酸点燃癫癎模型大鼠海马神经元保护作用研究

目的探讨生酮饮食对海人酸点燃癫癎模型大鼠海马神经元的保护作用。方法经海人酸制备SD大鼠癫癎模型,分别给予生理盐水+正常膳食(C组)、生理盐水+生酮饮食(K组)、海人酸+正常膳食(E组)和海人酸+生酮饮食(EK组),连续观察21 d后记录不同处理组大鼠体重、观察Ⅳ或Ⅴ级癫癎发作频率和持续时间,并通过HE染色和Nissl染色计数E组和EK组大鼠海马CA3区正常锥体神经元数目。结果 C组和K组大鼠均无癫癎发作,且海马CA3区锥体神经元数目正常。E组和EK组大鼠在观察过程中均出现Ⅳ或Ⅴ级癫癎发作,但EK组大鼠在饲养第21天时与E组相比,癫癎发作频率减少[(17.90±4.12)次对(30.50±4.40)次,P=0.000]、发作持续时间缩短[(212.70±17.75)s对(335.00±14.21)s,P=0.000],差异有统计学意义;EK组海马CA3区正常锥体神经元数目与E组相比增加[(117.67±7.51)个对(71.33±6.11)个,P=0.000],差异亦有统计学意义。结论生酮饮食对海人酸点燃癫癎模型大鼠海马神经元具有保护作用。     

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。     

Orexin受体拮抗剂对睡眠剥夺的戊四氮致(癎)大鼠癫(癎)发作及脑组织病理学变化的影响

目的 探讨orexin-1受体(OX1R)和orexin-2受体(OX2R)拮抗剂对睡眠剥夺(SD)的戊四氮(PTZ)致(癎)大鼠癫(癎)发作及脑组织病理学变化的影响.方法 雄性Wistar大鼠48只,随机分为正常对照(NC)组、PTZ组、SD+ PTZ( SD)组、SD+ PTZ+二甲基亚砜(DMSO)组、SD+ PTZ+ OX1R拮抗剂SB334867(SB)组和SD+ PTZ+ OX2R拮抗剂TCS OX229 (TCS)组.采用改良多平台SD法,SD前及SD 48 h分别给予相应组大鼠侧脑室注射DMSO、SB或TCS.SD 72 h给予各组腹腔注射PTZ 50 mg/kg诱导癫(癎)发作;观察各组大鼠癫(癎)发作的潜伏期、发作等级评分、发作持续时间及死亡率;应用常规染色法观察海马的病理学变化,免疫荧光法(BrdU标记)观察神经细胞增殖的变化.结果 (1)与PTZ组比较,SD组及DMSO组(癎)性发作的潜伏期明显缩短,发作等级评分、持续时间及死亡率明显增加(均P ＜0.001),海马CA3区神经元损害加重,海马齿状回门区和颗粒细胞下层BrdU阳性细胞数显著增多(P＜0.001);SD组与DMSO组间差异无统计学意义.(2)与SD组比较,SB组和TCS组大鼠(癎)性发作的潜伏期明显延长,发作等级评分、持续时间及死亡率明显下降(均P＜0.05),海马CA3区神经元损害明显减轻,齿状回门区和颗粒下层BrdU阳性细胞数减少(均P ＜0.05);TCS组的变化较SB组更显著(P＜0.05 ～0.01).结论 Orexin受体拮抗剂尤其是OX2R拮抗剂可通过减轻海马CA3区神经元的损害和抑制齿状回区细胞增殖减轻SD对PTZ诱导癫(癎)发作的不利影响.     

STAT3在匹罗卡品致(癎)大鼠星形胶质细胞增生中的作用

目的 探讨致(癎)状态下大鼠海马内信号转导与转录激活因子3(STA3)与星形胶质细胞增生的关系.方法 匹罗卡品(PILO)腹腔注射建立大鼠颞叶癫(癎)模型,免疫组织化学方法观察阻滞JAK/STAT通路前后大鼠海马p-STAT3与胶质纤维酸性蛋白(GFAP)阳性细胞的表达规律,双重免疫荧光方法观察p-STAT3与GFAP阳性细胞的关系.结果 癫(癎)发作3 h(SE 3 h)时即出现STAT3在海马内被激活,SE 3 d时达高峰,之后渐降低,至SE 30 d时仍维持在较正常时略高的水平上;GFAP阳性细胞数的变化规律与之类似.预先用AG490阻断STAT3通路后,海马区p-STAT3乃及GFAP阳性细胞数均明显减少.双重免疫荧光结果发现p-STAT3阳性胞核位于GFAP阳性细胞胞浆中.结论 匹罗卡品导致的癫(癎)伴有大鼠海马星形胶质细胞内STAT3的激活,STAT3的活化可能促进星形胶质细胞的反应性增生.     

戊四氮点燃癫癎大鼠空间学习记忆改变及海马NR2B表达

目的观察戊四氮点燃癫癎大鼠空间学习记忆功能变化及海马NMDA2型受体(NR2)B亚单位(NR2B)表达,探讨二者的关系及PTZ致癎大鼠认知障碍发生的分子机制。方法采用戊四氮(PTZ)慢性癫癎(CE)模型,Y-迷宫对两组大鼠进行行为学检测,免疫组织化学方法观察两组大鼠海马CA3区NR2B表达的变化,反转录多聚酶链反应(RT-PCR)方法检测大鼠海马NR2B mRNA的表达。结果癫癎组大鼠空间学习记忆能力受损;其海马CA3区NR2B阳性细胞较对照组明显减少(P<0.01),同时伴有海马NR2B mRNA表达下降(P<0.01)。结论戊四氮点燃癫癎大鼠空间学习记忆受损可能与海马神经元NR2B的表达减少有关。     

雌激素和克罗米酚对致癎大鼠海马γ-氨基丁酸免疫反应细胞和GABA_A受体α1亚单位表达的影响

目的了解雌激素和克罗米酚对海人藻酸(KA)致癎大鼠癫癎发作行为学的影响及其影响癫癎活动的部分机制。方法将去势的雌性大鼠添加雌激素(20mg/kg)或添加雌激素和克罗米酚治疗,比较各组大鼠致癎后癫癎发作的行为学变化;并采用间接免疫荧光法检测各组大鼠海马中γ-氨基丁酸(GABA)免疫反应细胞及GABAA受体α1亚单位表达的变化。结果添加雌激素治疗组大鼠癫癎发作的潜伏期和到达4/5级(4级或5级)的时间[分别为(24.63±11.44)min和(41.50±16.22)min]均较去势组[分别为(46.75±14.61)min和(65.13±12.99)min]明显缩短,而同时添加雌激素和克罗米酚治疗组的潜伏期[(43.50±5.75)min]比单纯添加雌激素组明显延长。雌激素组的阳性免疫反应细胞数在大鼠海马的某些区域也较去势组明显减少,克罗米酚组与雌激素组相比则有所增多。结论高水平的雌激素可促进癫癎发作,克罗米酚添加治疗具有一定的抗癫癎作用。这可能与脑内GABA能系统某些功能的改变有关。     

黄体酮对脑梗死大鼠缺血半暗带内IL-6的表达

目的观察黄体酮对大脑中动脉闭塞(MCAO)大鼠缺血半暗带内炎症反应的抑制作用以及脑梗死体积的影响。方法选择成年雄性S-D大鼠,体质量为250~300g,并将其随机分为假手术组、缺血组、溶剂治疗组、黄体酮治疗组。分别于6h、24h、72h三个时间点处死大鼠,取出脑组织,行免疫组化染色观察白介素-6(IL-6)的表达情况,并通过TTC染色检测脑梗死体积的变化。采用单因素方差分析,P0.05,差异有统计学意义。结果脑梗死发生后,缺血半暗带内IL-6的表达水平明显增高,经黄体酮治疗后,其表达水平明显下降(P0.05),脑梗死体积较治疗前亦明显缩小(P0.05)。结论黄体酮可以通过抑制大脑中动脉闭塞大鼠脑内炎症反应的程度,减轻梗死侧半球的脑损伤程度。     

Effects of allopregnanolone on sedation in men, and in women on oral contraceptives

Allopregnanolone is a known GABA(A) receptor agonist not previously given to men, or to women using oral contraceptives (OC). The effects of metabolites of sex hormones on the GABA(A) receptor are different between men and women. OC are known to change GABA(A) receptor subunit composition. These factors might play a role in the differential effect of allopregnanolone in men and women, and in women with or without OC. To study the sedative effect of and sensitivity to allopregnanolone in men and in women with OC, nine healthy men (mean age 24.6 years) and nine healthy women on OC (mean age 21.8 years) were given three, increasing, intravenous dosages (0.015, 0.03, and 0.045 mg/kg) of allopregnanolone. Saccadic eye velocity (SEV), subjective ratings, and electroencephalography (EEG) were used to evaluate the response to allopregnanolone. Repeated blood samples for analyses of serum allopregnanolone levels were drawn throughout the study day. Allopregnanolone decreased SEV more in women than in men, and increased subjective ratings of 'sedation'. The results in women on OC are similar to earlier results in women without OC. Subjective ratings of 'contentedness' decreased in men but increased in women. Serum levels of allopregnanolone were more highly increased in men compared to women. Other pharmacokinetic parameters were not different between sexes. On the EEG, beta power increased in men. In conclusion, men and women on OC reacted differently to allopregnanolone.     

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.     

The effect of B-vitamins on hyperhomocysteinemia in patients on antiepileptic drugs

Patients on antiepileptic drugs (AEDs) may have elevated levels of plasma total homocysteine (p-tHcy). The aim of this study was to assess the effect of B-vitamin supplementation on the levels of p-tHcy and markers of endothelial activation and lipid peroxidation. A total of 33 adult patients on AEDs were identified with either fasting (Group 1, n=23) or post methionine load (PML) (Group 2, n=10) hyperhomocysteinemia. Subjects were supplemented with B-vitamins for 30 days: folic acid 0.4 mg, pyridoxine 120 mg and riboflavin 75 mg per day. After supplementation, serum folate and pyridoxal phosphate had increased, while fasting and PML p-tHcy had decreased (P<0.0001) by 36 and 26%, respectively. Prior to supplementation, the Group 1 patients had elevated levels of P-selectin and von Willebrand factor (vWF) (P=0.05 and 0.03, respectively). After supplementation, the levels of intercellular cell adhesion molecules had decreased (P=0.01) and E-selectin decreased nonsignificantly (P=0.07). However, the levels of vascular cell adhesion molecules had increased (P<0.0001), while lipid peroxidation were unchanged. In conclusion, the combined supplementation with folic acid, pyridoxine and riboflavin reduced fasting and PML hyperhomocysteinemia in patients on AEDs. Patients with fasting hyperhomocysteinemia had elevated levels of P-selectin and vWF, which may indicate an increased risk of cardiovascular disease. Furthermore, B-vitamin supplementation influenced endothelial activation, although the clinical implication is uncertain.     

雌激素对实验性变态反应性脑脊髓炎大鼠发病影响的研究

目的探讨雌激素对实验性变态反应性脑脊髓炎(EAE)的影响。方法将30只大鼠随机分为EAE组及大、小剂量雌激素干预组,每组10只,制作EAE模型,大、小剂量雌激素干预组分别给予皮下注射苯甲酸雌二醇1 mg/kg/d、250μg/kg/d,连续10 d,观察各组的发病情况并采用HE染色观察脑和脊髓组织病理变化。结果大、小剂量雌激素干预组的临床症状均较EAE组轻,表现为发病率减少、潜伏期延长、进展期缩短、高峰期神经功能损害较轻。病理切片提示雌激素干预组脑和脊髓炎症细胞浸润明显减少,其中以大剂量组更明显。结论雌激素对多发性硬化动物模型EAE具有保护作用,且与剂量相关。     

Effect of hypercholesterolemia on thrombosis in rabbits

White mural thrombosis has been very rarely observed in the rabbit's aorta in experimental cholesterol atherosclerosis. Apparently cholesterol atherosclerosis by itself is not the appropriate damage or alteration required to trigger or initiate thrombosis in rabbits. It appeared to be of interest to determine if the hyperlipidemic state of the rabbit fed cholesterol could modify the amount of white thrombus formed after thrombosis was initiated by insertion of a standard length of polyethylene tube into the abdominal aorta. Twenty rabbits fed 0.5 gm cholesterol per day for 24 days developed severe hyperlipidemia. Polyethylene tubes were inserted into their abdominal aortae on the 24th day and rabbits were sacrificed on the 28th day. In this group, 61.1 mg ± 17.1 S.D. fresh wet weight of white thrombus or 17.0 mg ± 4.6 S.D. freeze-dried weight were found. In comparison, 20 normolipidemic control rabbits showed 67.2 mg ± 4.6 S.D. fresh weight or 15.7 ± 8.8 freeze-dried weight (P > 0.5). It is concluded that hypercholesterolemia induced by feeding cholesterol to rabbits does not modify the amount of white thrombus induced to form by a foreign plastic surface.     

新型抗癫痫药唑尼沙胺对点燃的影响

目的研究唑尼沙胺对大鼠杏仁核点燃及戊四唑(PTZ)化学性点燃模型的抗癫痫作用。方法建立大鼠杏仁核电刺激点燃、化学性点燃模型,探讨唑尼沙胺的抗点燃作用;测定唑尼沙胺对小鼠最大电休克的影响。结果唑尼沙胺50~200mg·kg-1ig可抑制大鼠杏仁核点燃发作,降低Racine's分级唑尼沙胺50,100mg·kg-1ig抑制PTZ化学性点燃发作,降低Ono分级(P<0.01)与OnoⅣ级发作百分率(P<0.01)。ZNS10.0mg·kg-1ig降低小鼠最大电休克诱发的,惊厥发生率(P<0.01)。结论唑尼沙胺对点燃发作具有拮抗作用。