APP5肽类似物165对老年性痴呆模型大鼠行为学及PSD95和Shank 1表达的影响

目的观察5肽类似物165对老年性痴呆（Alzhei mer disease,AD）模型大鼠学习记忆能力及突触后致密区蛋白95（postsynaptic density95,PSD95）和骨架蛋白Shank1表达的影响。方法将45只大鼠随机分为正常对照组、模型组和5肽类似物165治疗组,模型组和治疗组大鼠按体重3mg/kg行双侧侧脑室链脲佐菌素（streptozotocin,STZ）注射,第3天重复注射建立AD模型,对照组以人工脑脊液代替STZ。术后21d治疗组按体重0.34mg/（kg.d）给予APP5肽类似物165灌胃干预,其余两组以蒸馏水代替。3周后应用Morris水迷宫、免疫组织化学和Western blotting方法检测大鼠的学习记忆能力及PSD95和Shank1的表达。结果5肽类似物165治疗组大鼠的平均游泳时间较模型组明显缩短（P〈0.01）,且海马PSD95和Shank1阳性神经细胞数及PSD95和Shank1蛋白表达较模型组明显增加（P〈0.05）。结论5肽类似物165可显著提高大鼠学习记忆能力,增加大鼠海马PSD95和Shank1表达,表明其对突触功能和可塑性具改善作用。     

链脲佐菌素所致高血糖对大鼠缺血性脑损伤的影响

目的 研究链脲佐菌素所致糖尿病性高血糖（ＨＧ）对大鼠缺血性脑损伤的影响。方法 链脲佐菌素引起糖尿病４周的大鼠造成１０ｍｉｎ前脑缺血,观察缺血后脑组织的病理改变和癫痫发作情况。结果 在海马ＣＡ１区和下脚区,ＨＧ组与正常血糖（ＮＧ）组动物脑神经元损伤程度相似。然而,ＨＧ组动物有严重的顶叶皮质神经元坏死和“额外脑结构损伤”。ＨＧ组中４２．１％的动物脑缺血后出现癫痫发作,血浆葡萄糖浓度低于１２ｍｍｏｌ／Ｌ     

链脲佐菌素侧脑室注射后大鼠海马SYP改变

目的研究侧脑室内注射链脲佐菌素（streptozotocin,STZ）对大鼠海马突触素（synaptophysin,SYP）的影响。方法将40只SD大鼠随机分成对照组和模型组,于第1d和第3d侧脑室注射STZ3rag／kg建立大鼠AD模型,15d后进行Morris水迷宫试验,取海马后通过RT-PCR、western-blotting及免疫组织化学法检测SYP表达水平。结果与对照组相比较,侧脑室注射STZ后大鼠逃避潜伏期明显延长,穿越平台次数明显减少,SYP表达明显减少。结论侧脑室注射STZ后大鼠空间记忆力减退与突触结构受损有关。     

侧脑室注射链脲佐菌素对大鼠脑内能量代谢及线粒体功能影响的研究

目的探讨侧脑室注射链脲佐菌素(streptozotocin,STZ)对大鼠脑内能量代谢及线粒体功能的影响。方法 12只SD大鼠随机分为STZ组及生理盐水组。30天后电镜下观察两组大鼠海马及额顶颞皮层,并检测线粒体呼吸链复合物Ⅱ和Ⅳ、Na+ -K+ -ATP酶、活性,及三磷酸腺苷(adenosine triphosphate,ATP)、二磷酸腺苷(adeno- sine diphosphate,ADP)的浓度。结果 STZ组海马及额顶颢皮层电镜下有明显改变,ATP浓度、ATP/ADP比值及线粒体功能降低明显(P>0．05),ADP浓度升高明显(P>0．05)。结论侧脑室注射STZ引起能量代谢明显异常及线粒体活性的下降并有神经细胞受损。     

链脲佐菌素剂量对C57BL/6J小鼠糖尿病神经病理性疼痛诱导效应的影响

目的观察不同剂量链脲佐菌素(STZ)对C57BL/6J小鼠糖尿病神经病理性痛诱导效应的影响,探讨其量效关系及最佳剂量。方法雄性C57BL/6J小鼠85只,随机分为正常对照组(n=10)和A～E 5个实验组(n=15,STZ腹腔注射剂量分别为80、100、120、140、160mg/kg),对照组腹腔注射等体积柠檬酸缓冲液。观察各组血糖、机械痛阈值、热刺痛阈值和28d生存率的变化,分析其与STZ剂量的关系。结果 A～E各组糖尿病成模率均>86.7%。C组神经病理性疼痛成模率(66.7%)均高于其余各组;E组28d生存率(46.7%)与A、B组间均存在显著差异(P<0.05)。结论 C57BL/6J小鼠腹腔注射STZ以120mg/kg为最佳剂量。     

链脲佐菌素对人神经母细胞瘤系SH—SYSY细胞生长的影响

目的 研究体外链脲佐菌素(STZ)对人神经母细胞瘤(SH-SY5Y)细胞生长以及SH-SY5Y细胞胰岛素信号转导通路相关蛋白表达的影响.方法 采用MTT测定法测定SH-SY5Y细胞活性,乳酸脱氢酶(LDH)漏出率测定法观察SH-SY5Y细胞生长情况;应用Western blot法检测胰岛素信号转导通路相关蛋白胰岛素受体-1(IRS-1)、磷脂酰肌醇激酶-3(PI3K)等的改变.结果 STZ与SH-SY5Y细胞共同孵育可抑制SH-SY5Y细胞生长,阻断胰岛素对SH-SY5Y细胞的促生长作用,且STZ抑制细胞生长呈明显的时间-剂量效应.随STZ浓度增加,LDH漏出率也增加.Western blot半定量分析发现IRS-1、PI3K表达减少.结论 体外STZ与SH-SY5Y细胞共孵育可能影响胰岛素/胰岛素样生长因子-1信号转导系统对细胞的促生长作用.STZ与SH-SY5Y细胞共孵育可能作为体外胰岛素信号转导通路的一种细胞模型应用于某些神经药理学研究.     

链脲佐菌素型糖尿病模型大鼠股骨头坏死的血脂、凝血代谢及组织病理学变化

背景：糖尿病性股骨头坏死的病理机制和病变特征至今尚未阐明。有研究证实，骨坏死与低纤溶状态有关，高脂血症是引起股骨头缺血性坏死的重要诱因。 目的：拟观察糖尿病股骨头缺血性坏死模型大鼠的血脂、凝血代谢及组织病理学变化。 设计、时间及地点：随机对照动物实验，于2006-04/2007-10在西安交通大学医学院第二附属医院骨科教研室及西安交通大学医学院病理教研室完成。 材料：选用12周龄SD大鼠100只，适应性喂养2周后，精确称重，按性别抽取24只(雌雄各半)作为空白对照组，其余76只行速发型糖尿病大鼠模型制备。 方法：模型组大鼠将60 mg/kg链脲佐菌素腹腔快速推入，建立速发型糖尿病大鼠模型，使动物进食及饮水时双后腿负重，模拟人类髋关节生理状态。对照组24只给予腹腔针刺，不注入任何液体。 主要观察指标：两组大鼠分别于造模成功后第4，8，12，18周麻醉后分批处死，对照组每次随机处死6只，模型组每次处死18只，进行大体形态、组织病理学、血脂及凝血指标检测。 结果：造模成功后模型组大鼠日渐消瘦，食欲旺盛，饮水量增加，体质量逐渐下降。模型组大鼠电镜下均可见骨小梁表面成骨细胞减少，活性降低；光镜下观察，与同期对照组比较，自造模第4周后空骨陷窝计数显著增加(P < 0.01)。与同期对照组相比，模型组大鼠造模后第8，12，18周活化部分凝血活酶时间、凝血酶原时间及凝血酶时间显著缩短(P < 0.05~0.01)，纤维蛋白原水平显著升高 (P < 0.05~0.01)，血清总胆固醇、三酰甘油浓度显著升高(P < 0.01)，高密度脂蛋白显著降低 (P < 0.05~0.01)。 结论：糖尿病可引起长期骨质疏松、高脂血症、血液高凝状态，这些因素共同作用下将会最终导致糖尿病股骨头缺血性坏死的发生。     

APP17肽对APP转基因小鼠学习记忆能力和海马神经细胞凋亡的影响

目的观察APP(-βamyloid precursop protein,APP)17肽(APP695中319-335肽段)对APP转基因小鼠(APP695V717I)学习、记忆能力及海马神经细胞凋亡的影响。方法3月龄的APP695转基因小鼠随机分为模型组和APP17肽治疗组,正常对照组采用月龄和性别与之相匹配的C57BL/6J小鼠。APP17肽治疗组给予皮下注射APP17肽,每只每次0.34μg,每周3次;模型组和正常对照组给予等体积NS。应用水迷宫试验观察小鼠学习、记忆功能的变化,以流式细胞技术分析海马神经细胞凋亡率和线粒体膜电位的变化。结果(1)水迷宫试验结果显示,模型组小鼠存在明显学习和记忆功能障碍,其第3、4、5天游完全程的时间[(93.22±16.35)、(86.73±20.26)、(77.13±29.35)s]和错误反应次数[(6.63±2.16)、(5.81±2.13)、(5.33±1.41)次]均较正常对照组[分别为(70.89±20.19)、(61.25±21.88)、(54.63±16.92)s和(5.01±1.93)、(2.97±0.96)、(2.31±1.01)次]增多(P<0.05);APP17肽治疗组小鼠的行为学障碍明显轻于模型组(P<0.05),其上述水迷宫检测结果与正常对照组比较差异无统计学意义。(2)与正常对照组小鼠海马神经细胞凋亡发生率[(3.13±1.19)%]和线粒体膜电位[(176.39±13.88)mV]比较,模型组凋亡发生率[(8.06±2.31)%]显著增加(P<0.01)而线粒体膜电位[(97.51±15.73)mV]明显降低(P<0.01);APP17肽治疗组检测结果与正常对照组接近,海马神经细胞凋亡发生率[(4.38±1.26)%]低于模型组(P<0.01),线粒体膜电位[(168.35±19.29)mV]高于模型组(P<0.01)。结论APP695转基因小鼠存在学习和记忆功能障碍,且其海马神经细胞凋亡率增加,线粒体膜电位降低;APP17肽能够明显改善该转基因小鼠的学习和记忆能力,其作用机制可能是通过稳定线粒体膜电位及抑制凋亡发生而实现。     

慢性强迫游泳应激对大鼠海马神经元再生和p-CREB表达的影响

目的 研究慢性强迫游泳应激模型大鼠海马神经元再生和磷酸化环磷酸腺苷反应元件结合蛋白（p-CREB）的表达.方法 30只雄性SD大鼠随机分为3组：强迫游泳7 d组（S1组）、强迫游泳14 d组（S2组）和对照组.S1组和S2组分别连续强迫游泳7 d和14 d,每天5 min,水温（10±0.5）℃.采用免疫组化半定量测定大鼠海马5-溴脱氧尿苷（BrdU）和p-CREB阳性细胞表达情况.结果 免疫组化结果 显示,在整个海马结构中BrdU及p-CREB的阳性细胞主要集中于齿状回的颗粒细胞下层.与对照组比较,S1组、S2组大鼠海马齿状回BrdU和p-CREB阳性细胞数均明显减少（P＜0.01）;而与S1组比较,BrdU阳性细胞数无统计学差异（P＞0.05）,S2组p-CREB阳性细胞数进一步减少（P＜0.01）.结论 慢性强迫游泳应激可导致海马神经元再生功能障碍,其机制可能与p-CREB信号转导通路有关. 基金项目：     

次声对大鼠海马5-HT、5-HTR、RyR表达及恢复的研究

目的研究大鼠海马5-羟色胺(5-HT)、5-羟色胺受体(5-HTR)、兰尼定受体(RyR)表达在8 Hz、130 dB的次声作用后1周和2周时的恢复情况. 方法大鼠暴露于8 Hz、130 dB次声1、7、14、21、28、35、42 d后置安静环境观察1周或2周,取脑组织并进行免疫组织化学染色,光学显微镜下观察海马5-HT、5-HTR、RyR表达. 结果次声作用组大鼠脑组织海马5-HT、5-HTR、RyR表达均较对照组减少,最低值分别出现于21 d、28 d和42 d(P<0.01).次声停止作用后,5-HT、5-HTR、RyR表达均逐渐回升,且随时间延长大部分恢复到正常对照水平.结论 8 Hz、 130 dB次声可引起大鼠海马5-HT、5-HTR、RyR表达减少,其变化规律与观察指标有关;次声引起的5-HT、5-HTR、RyR表达减少在停止次声作用后可逐渐恢复正常.     

糖尿病大鼠海马组织中周期素依赖性激酶5表达的实验研究

目的：分析糖尿病大鼠海马组织中周期素依赖激酶5（CDK5）蛋白表达情况，探讨糖尿病影响学习与记忆的机制。方法：Y-迷宫评估大鼠学习与记忆成绩，用免疫组化法和Western blot检测海马区CDK5蛋白表达。结果：DM组大鼠第4周迷宫作业错误反应次数和全天总反应时间分别为（3．43±2．20）次、（115．42±36．64）s，对照组为（2．17±1．34）次，（90．83±20．27）s；CDK5蛋白表达相对水平为15．62±7．07，对照组为20．61±5．63，两组间差异有统计学意义（P〈0．05）。结论：糖尿病大鼠CDK5蛋白表达水平下降，学习记忆能力下降，提示CDK5可能参与了学习记忆过程。     

Effects of Jiawei Wendan decoction on hippocampal p-CREB protein expression in a rat model of depression☆

BACKGROUND： Jiawei W＇endan decoction can elevate hippocampal brain-derived neurotrophic factor （BDNF） protein expression in rats with depression. It has been hypothesized that Jiawei Wendan decoction can exhibit antidepressant effects through the hippocampal signal transduction pathway of cyclic adenosine monophosphate response element binding protein （CREB）-BDNF. OBJECTIVE： Using phosphorylated-CREB （p-CREB） as an entry point, the present study was designed to observe intervention effects ofJiawei Wendan decoction compared with fluoxetine. DESIGN, TIME AND SETTING： A randomized, controlled, cellular biology experiment was performed at the Central Laboratory of Guangxi University of Traditional Chinese Medicine, MATERIALS： A total of 40 healthy, male, Sprague-Dawley rats were included in the present study. Rhizoma Acori Talarinowii （Shichangpu）, Flos Albiziae （Hehuanhua）, Rhizoma Pinelliae （Banxia）, Caulis Bambusae in Taeniam （Zhuru）, Fructus Aurantii Immaturus （Zhishi）, Poria （Fuling）, and Radix Bupleuri （Chaihu）, the primary ingredients ofJiawei Wendan decoction, were purchased from First Affiliated Hospital of Guangxi University of Traditional Chinese Medicine. The raw drug was decocted at a concentration of 1.5 g/mL. Fluoxetine capsules were purchased from Shanghai Zhongxi Pharmaceutical Co., Ltd., China. METHODS： Following behavioral testing, 36 rats were selected from the initial 40 rats according to similar behavioral scores, and were randomly divided into 4 groups： model （n = 8）, Jiawei Wendan decoction-treated （n = I 0）, fluoxetine-treated （~ = 10）, and normal control （n = 8）. All rats, except for those in the normal control group, were separately raised in a chronic and unpredictable, mild-stimulation environment for 2 l days to establish a depression model. The Jiawei Wendan decoction-treated and fluoxetine-treated groups were intragastrically administered Jiawei Wendan decoction （12 g/kg/d） and fluoxetine （1.8 mg/kg/d）, resp     

Effects of Jiawei Wendan decoction on hippocampal p-CREB protein expression in a rat model of depression

BACKGROUND: Jiawei Wendan decoction can elevate hippocampal brain-derived neurotrophic factor (BDNF) protein expression in rats with depression. It has been hypothesized that Jiawei Wendan decoction can exhibit antidepressant effects through the hippocampal signal transduction pathway of cyclic adenosine monophosphate response element binding protein (CREB)-BDNF. OBJECTIVE: Using phosphorylated-CREB (p-CREB) as an entry point, the present study was designed to observe intervention eftects ofJiawei Wendan decoction compared with fluoxetine. DESIGN, TIME AND SETTING: A randomized, controlled, cellular biology experiment was performed at the Central Laboratory of Guangxi University of Traditional Chinese Medicine. MATERIALS: A total of 40 healthy, male, Sprague-Dawley rats were included in the present study. Rhizoma Acori Talarinowii (Shichangpu), Flos Albiziae (Hehuanhua), Rhizoma Pinelliae (Banxia), Caulis Bambusae in Taeniam (Zhuru), Fructus Aurantii Immaturus (Zhishi), Poria (Fuling), and Radix Bupleuri (Chaihu), the primary ingredients ofJiawei Wendan decoction, were purchased from First Affiliated Hospital of Guangxi University of Traditional Chinese Medicine. The raw drug was decocted at a concentration of 1.5 g/mL. Fluoxetine capsules were purchased from Shanghai Zhongxi Pharmaceutical Co., Ltd., China. METHODS: Following behavioral testing, 36 rats were selected from the initial 40 rats according to similar behavioral scores, and were randomly divided into 4 groups: model (n = 8), Jiawei Wendan decoction-treated (n = 10), fluoxetine-treated (n = 10), and normal control (n = 8). All rats, except for those in the normal control group, were separately raised in a chronic and unpredictable, mild-stimulation environment for 21 days to establish a depression model. The Jiawei Wendan decoction-treated and fluoxetine-treated groups were intragastrically administered Jiawei Wendan decoction ( 12 g/kg/d) and fluoxetine ( 1.8 mg/kg/d), respectively. The model and normal control groups received double-distilled water (2 mL/kg/d) once a day for a total of 21 days. MAIN OUTCOME MEASURES: Prior to depression induction and subsequent to administration, memory and degree of depression were measured by open field and tail suspension tests, respectively. At 24 hours after administration, p-CREB expression in the hippocampal CA3 region was analyzed by the immunohistochemical SABC method. RESULTS: Two rats from the Jiawei Wendan decoction-treated group, and one from the fluoxetine-treated group, died of unknown causes. The remaining rats in each group, except for one rat that was randomly selected from the fluoxetine-treated group, were included in the final analysis. Following administration, the model group exhibited significantly decreased numbers of hippocampal p-CREB-positive cells, horizontal and vertical movements, and significantly prolonged duration of immobility in tail-suspension test, compared with the normal control group (P < 0.01 ). The numbers of p-CREB-positive cells, as well as horizontal and vertical movements, were significantly greater, and the duration of immobility was significantly shorter, in the Jiawei Wendan decoction and fluoxetine groups, compared with the model group (P < 0.05-0.01 ). There was no significant difference in the above-mentioned three indices between the Jiawei Wendan decoction and fluoxetine groups (P>0.05). CONCLUSION: Jiawei Wendan decoction increased hippocampal p-CREB expression by influencing the rat hippocampal signal transduction pathway of CREB-BDNF. Jiawei Wendan decoction exhibited antidepressant effects equivalent to fluoxetine.     

托吡酯长期应用对癫痫幼鼠学习记忆及海马NMDAR1 mRNA表达的影响

目的:研究托吡酯(TPM)长期应用对癫癎幼鼠学习记忆功能的影响,并观察海马NMDAR1 mRNA表达的变化。方法:以锂-匹罗卡品制作癫癎模型,TPM灌胃,5周后使用Morris水迷宫评价大鼠的学习记忆功能,通过核酸原位杂交法观察海马各区NMDAR1mRNA表达的变化。结果:癫癎幼鼠的学习记忆功能较正常幼鼠明显下降;癫癎幼鼠学习记忆功能经中、小剂量TPM干预后未发生变化,而经大剂量TPM于预后则明显下降;大鼠海马各区NMDAR1 mRNA的表达在TPM干预后无明显变化。结论:癫癎持续状态(SE)后的慢性癫癎对幼鼠的视觉空间学习记忆功能造成损害,长期大剂量应用TPM可加重这一损害,该损害可能与海马CA1、CA3区NMDAR1 mRNA的表达无关。     

行为学训练对海马损伤梗死大鼠齿状回区BrdU和Nestin表达的影响

目的 探讨行为学训练对海马损伤梗死大鼠齿状回区神经干细胞增殖的影响。方法 采用光化学法制成单侧海马损伤梗死模型大鼠72只,随机分为训练组（n=36）和自由活动组（n=36）,每个组设1、7、14、21、28及35d 6个亚组。另设正常对照组36只,与模型组对应分为1、7、14、21、28及35d 6个亚组。训练组大鼠于造模1d后给予水迷宫训练,自由活动组大鼠自由活动,不予水迷宫训练。免疫荧光双标记法观察各不同时间点大鼠海马齿状回区溴脱氧尿嘧啶核苷（Bromodeoxyuridine,BrdU）与巢蛋白（Nestin）的双标记表达情况。结果 正常对照组大鼠海马齿状回区有少量BrdU/Nestin双标记阳性细胞,训练组及自由活动组大鼠在7、14、21及28d海马损伤梗死侧齿状回区BrdU/Nestin双标记阳性细胞数量均有显著增多（P <0.01）;训练组大鼠7、14、21、28d时海马损伤梗死侧齿状回区的BrdU/Nestin双标记阳性细胞数量显著高于自由活动组（P <0.01）;至35d时,训练组及自由活动组大鼠海马损伤梗死侧齿状回区BrdU/Nestin双标记阳性细胞数量与正常对照组无明显差异（P >0.05）。结论 行为学训练能显著增强海马损伤梗死大鼠齿状回区神经干细胞的增殖,促进神经功能恢复。     

Protection of Vasoactive Intestinal Peptide on the Blood-Brain Barrier Dysfunction Induced by Focal Cerebral Ischemia in Rats

ObjectiveTo investigate the effects of vasoactive intestinal peptide on the blood brain barrier function after focal cerebral ischemia in rats.Materials and methodsRats were intracerebroventricular injected with vasoactive intestinal peptide after a two hours middle cerebral artery occlusion. Functional outcome was studied with the neurological severity score. The brain edema and the infarction were evaluated via histology. The blood brain barrier permeability was assessed using Evans Blue dye injection method. We also measure the apoptosis of brain microvascular endothelial cells and brain levels of B-cell leukemia-2 protein by immunohistochemical analysis and western blotting, respectively.ResultsIn contrast to the cases treated with vehicle at 72 h after middle cerebral artery occlusion, the treatment with vasoactive intestinal peptide significantly (P < 0.05) reduced the neurological severity score, the brain edema and infarct volume. The Evans Blue leakage and brain water content were obviously reduced (P < 0.05) in vasoactive intestinal peptide-treated rats compared with those of control rats at 72 and 96 h after stroke. In addition, vasoactive intestinal peptide decreased the numbers of terminal deoxynucleotidyl transferase-mediated dUTP-nick end labeling positive endothelial cells and increased the protein levels of B-cell leukemia-2 in the ischemic hemisphere at 72 h after ischemia.ConclusionsOur data suggest that treatment with vasoactive intestinal peptide ameliorates the blood brain barrier function, contributing to reduce in brain damage both morphologically and functionally in the ischemic rat. This amelioration may be associated with attenuation in apoptosis of brain microvascular endothelial cells by increased B-cell leukemia-2 expression.