3-硝基丙酸诱导继发性肌张力障碍大鼠模型的行为学研究

目的 观察3-硝基丙酸(3-NP)诱导的继发性肌张力障碍大鼠模型的行为学变化,以建立一个可靠的继发性肌张力障碍动物模型.方法 36只雄性SD大鼠按随机数字表法分为3组,每组各12只,前2组为实验组(低剂量组立体定向注射2000 nmol 3-NP至大鼠左侧尾壳核,高剂量组立体定向注射4000nmol 3-NP),第3组为对照组(注射等体积生理盐水).术后观察大鼠头部扭转和自主行为变化,平板实验观察大鼠通过时间及步幅.结果 实验组大鼠给药后开始出现姿势障碍,头部向同侧扭转,其中高剂量组扭转评分明显大于低剂量组,比较差异有统计学意义(P＜0.05).平板实验结果显示:实验组大鼠通过时间较对照组明显延长,步幅较对照组明显变小,比较差异有统计学意义(P＜0.05),且偏于木板一侧行走,症状随剂量增加而加重.对照组大鼠术后第2天基本恢复术前状态,无头部偏转,均顺利通过木板实验.结论 大鼠尾壳核注射3-NP可诱导产生稳定的继发性肌张力障碍动物模型.     

鱼藤酮帕金森病大鼠模型建立的研究

目的探讨鱼藤酮帕金森病大鼠模型的建立方法.方法采用背部皮下注射鱼藤酮,然后观察大鼠的行为学及黑质-纹状体的酪氨酸羟化酶免疫活性变化、海马病理学变化.结果经过1～8周观察,鱼藤酮处理大鼠出现明显的行为学、黑质-纹状体酪氨酸羟化酶免疫活性变化,海马无明显病理学变化,模型显示出帕金森病的典型特征.结论背部皮下注射鱼藤酮的方法可成功制作帕金森病大鼠模型.     

抑郁状态下大鼠海马Notch1信号系统的改变

目的 了解大鼠抑郁模型中海马神经重塑障碍与Notch1信号系统功能改变的关系.方法 54只大鼠随机分为CUMS 14 d组、CUMS 28 d组和对照组,前两组接受慢性不可预知温和应激和孤养(chronic unpredictable mild stress,CUMS)14 d和28 d建立抑郁模型.采用免疫组化、免疫荧光、RT-PCR和Western blot 法.测定大鼠海马神经干细胞的增殖、存活和分化以及Notch1信号通路各个因子的基因及蛋白表达水平的改变.结果 与对照组比较,CUMS 14 d组和CUMS 28 d组大鼠海马神经干细胞增殖与存活明显减少(P<0.001).CUMS 28 d组大鼠海马神经干细胞分化NeuN/BrdU、GFAP/BrdU比例无明显差异(P>0.05).与对照组比较.CUMS 14 d组和CUMS 28 d组Notch1信号通路各因子(NICD、Hes1、Hes5和Jag1)基因表达和蛋白水平明显降低(P<0.01).结论 抑郁大鼠海马齿状回神经干细胞增殖和存活受到抑制,但分化无改变;同时,大鼠海马Notch1功能下调.提示Notch1信号系统可能与抑郁症海马神经再生障碍有关.     

慢性应激抑郁模型大鼠海马GSK-3β的表达

目的 研究慢性应激对大鼠海马糖原合成酶激酶-3β(GSK-3β)表达的影响.方法 实验组大鼠通过21d的应激刺激制作抑郁动物模型,此期间对照组大鼠正常饲养.Open-field法检测行为学改变,Western blot法测定大鼠海马GSK-3β的表达.结果 慢性应激后,实验组大鼠体重、水平穿越格数、直立次数和修饰次数均显著低于对照组(P<0.01);实验组大鼠海马GSK-3β表达的整合光密度值(28413.30±2501.31)高于对照组(23149.50±3107.25),差异有统计学意义(P<0.05).结论 慢性应激使大鼠海马GSK-3β的表达上调.     

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。     

β-淀粉样蛋白诱导大鼠海马神经元凋亡及海马亲环素A表达的变化

目的 探讨Aβ25-35对大鼠海马神经元的损伤及对亲环素A(CyPA)表达的影响. 方法 健康Wister大鼠60只按照随机数字表法分为实验组(n=30)和对照组(n-30),实验组大鼠采用Aβ25-35注射入双侧海马制造阿尔茨海默病的动物模型,对照组大鼠同样位置注射入等量生理盐水.HE染色观察2组大鼠海马CA1区神经元形态,TUNEL染色检测细胞凋亡,RT-PCR检测CyPAmRNA的表达,Western blotting检测CyPA蛋白的表达. 结果 实验组大鼠海马CA1区神经元遭到破坏,细胞凋亡增加,数量减少,且随时间延长细胞凋亡情况进一步加重,造模后1d、7d、14d较对照组比较差异均有统计学意义(P＜0.05).实验组大鼠海马组织CyPA mRNA和CyPA蛋白表达量随时间呈现出先增长后降低的趋势,1d、7d时明显高于对照组,差异均有统计学意义(P＜0.05).14d时明显低于对照组大鼠,其中CyPA蛋白的差异具有统计学意义(P＜0.05). 结论 Aβ25-35通过加重大鼠海马神经元凋亡诱发神经毒性作用,CyPA表达的增加可能是一种内源性保护因素.     

帕金森病临床前期小鼠模型的建立

目的建立帕金森病临床前期的小鼠动物模型,了解帕金森病发病前的组织病理学改变,特别是小鼠中脑黑质、纹状体的超微病理及突触数量的变化。方法60只C57Bl/6j小鼠,采用小剂量MPTP多次慢性给药方法(4mg/kg.d)建立帕金森病临床前期小鼠动物模型。通过光学显微镜及电子显微镜观察小鼠黑质及纹状体的超微病理变化;利用色谱分析测定模型小鼠纹状体的多巴胺含量;通过动物爬杆试验(pole test)检测小鼠肢体运动协调的行为学改变。结果鼠脑黑质部分神经细胞变性及纹状体内突触数量减少;各给药组小鼠纹状体内多巴胺的含量均有不同程度的下降,同时各实验组小鼠行为学的定量及定性观察均未出现明显的行为学改变。结论低剂量MPTP慢性给药方式可以建立帕金森病临床前期的小鼠动物模型。该模型的超微形态学、生物化学及动物的行为学等结果可以作为研究帕金森病发病早期的参考指标。     

粒细胞集落刺激因子对血管性痴呆大鼠海马神经细胞凋亡的影响

背景：研究发现，粒细胞集落刺激因子可激活脑内成体神经干细胞，刺激其增殖和分化，还能促进脑内各种神经营养因子分泌，减小脑缺血动物模型的缺血灶，促进慢性脑卒中模型缺失神经功能的长期恢复。 目的：观察粒细胞集落刺激因子对血管性痴呆大鼠海马神经细胞凋亡的作用。 方法：采用永久性双侧颈总动脉结扎法建立大鼠血管性痴呆模型，抽签法随机分为4组：实验组(皮下注射粒细胞集落刺激因子干预治疗)、对照组(注射生理盐水)、假手术组(仅行颈前正中切开，不结扎颈总动脉)。采用Morris水迷宫进行定向航行观察大鼠逃避潜伏期，评价大鼠空间学习记忆能力，TUNEL染色和图像分析大鼠海马神经细胞的凋亡。 结果与结论：对照组和实验组大鼠脑缺血后7 d，大鼠平均逃避潜伏期较假手术组明显延长(P < 0.01)，海马组织TUNEL阳性凋亡细胞较假手术组显著增高(P < 0.01)，随着时间的延长，14，28 d时大鼠学习记忆功能障碍逐渐加重，相应海马组织TUNEL阳性凋亡细胞逐渐增加。14，28 d时实验组大鼠平均逃避潜伏期比对照组明显缩短，海马组织TUNEL阳性凋亡细胞也较对照组显著减少。说明脑缺血后早期给予外源性粒细胞集落刺激因子有助于减少海马组织神经细胞的凋亡，改善大鼠学习记忆能力。     

TauN368和磷酸化α-synuclein在3种帕金森病动物模型脑区的表达

目的探讨3种不同帕金森病(Parkinson disease,PD)动物模型黑质、纹状体和海马中磷酸化α-synuclein(S129)[以下简称为p-α-syn(S129)]和天冬酰胺内切酶(asparagine endopeptidase,AEP)特异性切割产生的tauN368片段的表达水平。方法建立慢性1-甲基-4-苯基-1,2,3,6-四羟吡啶(MPTP)诱导的PD小鼠模型(n=10)、鱼藤酮诱导的小鼠PD模型(n=10)以及鱼藤酮诱导的大鼠PD模型(n=10)共3种模型,每种模型均设立对照组(n=10)。采用酪氨酸羟化酶(tyrosine hydroxylase,TH)免疫染色评估不同PD动物模型黑质多巴胺能神经元损伤情况,采用Western blot方法检测各组动物中脑黑质、纹状体、海马p-α-syn(S129)和tauN368的表达。结果 3种PD动物模型中脑黑质致密部TH阳性神经元数目均较对照组明显减少,提示造模成功。Western blot检测结果显示,MPTP诱导的小鼠PD模型黑质、纹状体及海马tauN368表达均较对照组升高(P0.05),黑质和海马p-α-syn(S129)表达无统计学差异(P0.05);鱼藤酮诱导的小鼠PD模型黑质及纹状体p-α-syn(S129)和tauN368表达均较对照组增加(P0.05),而海马p-α-syn(S129)和tauN368表达与对照组比较均未见统计学变化(P0.05);鱼藤酮诱导的大鼠PD模型黑质、纹状体及海马p-α-syn (S129)表达较对照组增加(P0.05),而黑质和海马tauN368的表达无统计学差异(P0.05)。结论鱼藤酮诱导的小鼠PD模型黑质和纹状体同时存在tauN368和p-α-syn(S129)的差异性表达,提示该模型可能是研究PD发病中tauN368和α-synuclein相互作用机制的理想动物模型。     

多巴胺失调综合征

<正>帕金森病(Parkinson’s disease,PD)是一种常见的中枢神经系统变性疾病,平均发病年龄为60岁左右。最主要的病理改变是中脑黑质多巴胺能神经元的变性死亡,由此而引起纹状体多巴胺(DA)含量显著性减少。临床主要表现为静止性震颤、运动迟缓、肌强直和姿势步态障碍及其他非运动     

谷氨酸能低下精神分裂症模型大鼠不同脑区核因子-кB表达分析

目的 了解地卓西平马来酸盐[Dizocilpine maleate,MK-801]所致谷氨酸能低下精神分裂症大鼠前额皮质、海马、纹状体核因子-кB(Nuclear factor-кB,NF-кB)基因的表达,探讨其参与精神分裂症的病理机制.方法 12只雄性Spragus-Dawley大鼠随机分为模型组和对照组,前者给予腹腔注射Mк-801(0.8 mg/kg)建立精神分裂症模型,后者只注射生理盐水.采用小动物行为分析系统记录大鼠90 min内自发活动,采用TransAM技术检测大鼠前额皮质、海马、纹状体的NF-кB p65亚基蛋白活性,半定量PCR检测NF-кB p65亚基基因mRNA的表达.结果 ①与对照组相比,模型组大鼠白发活动增加(P＜0.05),并有典型的刻板行为和共济失调,表现出了类似精神分裂症的行为.②模型组前额皮质、海马、纹状体的NF-кB p65亚基蛋白活性均低于对照组,差异均具有统计学意义(P＜0.05).③模型组前额皮质、海马、纹状体3个脑区NF-кB p65亚基mRNA表达均不同程度高于对照组相应脑区,但只有在前额皮质和纹状体两组之间的差异具有统计学意义(P＜0.05).结论 谷氨酸能低下精神分裂症模型人鼠前额皮质、海马、纹状体NF-кB的活性受到了抑制,而mRNA表达上凋,提示NF-кB可能参与了精神分裂症的病理过程.     

Beta-endorphin and methionine-enkephalin levels in discrete brain regions, spinal cord, pituitary gland and plasma of morphine tolerant-dependent and abstinent rats

The effect of morphine tolerance-dependence, protracted and naloxone-precipitated abstinence on the levels of beta-endorphin and methionine-enkephalin in discrete brain regions, spinal cord, pituitary gland and plasma was determined in the male Sprague-Dawley rats. Among the brain regions examined, the levels of beta-endorphin in descending order were: hypothalamus, amygdala, midbrain, hippocampus corpus striatum, pons and medulla and cortex. The levels of beta-endorphin in midbrain, hypothalamus, and pituitary of morphine tolerant-dependent rats were decreased significantly. During protracted withdrawal beta-endorphin levels were decreased in amygdala, spinal cord and pituitary. During naloxone-precipitated abstinence beta-endorphin levels were increased in corpus striatum, midbrain and cortex. In addition, in naloxone-precipitated abstinence beta-endorphin levels were decreased in pituitary gland and hippocampus but increased in plasma. The levels of methionine-enkephalin in brain regions in decreasing order were: corpus striatum, pons and medulla, amygdala, hypothalamus, midbrain, hippocampus and cortex. The levels of methionine-enkephalin in pons and medulla, amygdala, hippocampus and pituitary gland were decreased in morphine tolerant-dependent rats. During protracted abstinence from morphine, methionine-enkephalin levels in spinal cord, amygdala, pons and medulla, midbrain, cortex, corpus striatum and pituitary gland were decreased. The levels of methionine-enkephalin in hypothalamus and corpus striatum of naloxone-precipitated abstinent rats were increased but were decreased in amygdala and pituitary gland. These results suggest that during morphine tolerance-dependence and during protracted abstinence beta-endorphin and methionine-enkephalin levels in discrete brain regions and pituitary gland are decreased.(ABSTRACT TRUNCATED AT 250 WORDS)     

慢性应激大鼠中枢5-羟色胺含量及色氨酸羟化酶-2的表达变化

目的 探讨慢性不可预知轻度刺激（CUMS）大鼠模型中枢5-羟色胺（5-HT）含量和色氨酸羟化酶-2（TPH-2）的表达变化以及抗抑郁药的影响.方法 24只大鼠被随机分成3组,每组8只,A组为对照组,不给予刺激;B组为刺激不给药组,即CUMS应激鼠;C组为刺激给药组,即CUMS应激＋西酞普兰治疗组.实验为期6周,每周为大鼠称重,每3周测大鼠的糖水偏爱性,造模前后通过旷场试验评价大鼠行为,6周后对大鼠进行处死.处死后脑组织取样测定各脑区5-HT浓度及TPH-2 mRNA的表达水平.结果 经过6周干预,与A组比较,B组和C组体重明显降低（P＜0.05）;与A组及C组比较,B组糖水偏爱度显著降低（P＜0.05）,同时行为增多;B组5-HT浓度在海马中高于A组和C组,差异有统计学意义（P＜0.05）;在纹状体中,B组和C组5-HT浓度低于A组,差异有统计学意义（P＜0.05）;B组大鼠TPH2 mRNA在中缝核中表达低于A组,差异有统计学意义（P＜0.05）.结论 慢性应激可能引起中枢海马5-HT浓度增高及中缝核TPH-2表达降低,抗抑郁药可能引起中枢海马5-HT浓度降低,但未能影响中枢TPH-2的表达.     

β-endorphin and methionine-enkephalin levels in discrete brain regions, spinal cord, pituitary gland and plasma of morphine tolerant-dependent and abstinent rats

The effect of morphine tolerance-dependence, protracted and naloxone-precipitated abstinence on the levels of β-endorphin and methionine-enkephalin in discrete brain regions, spinal cord, pituitary gland and plasma was determined in the male Sprague-Dawley rats. Among the brain regions examined, the levels of β-endorphin in descending order were: hypothalamus, amygdala, midbrain, hippocampus, corpus striatum, pons and medulla and cortex. The levels of β-endorphin in midbrain, hypothalamus, and pituitary of morphine tolerant-dependent rats were decreased significantly. During protracted withdrawal β-endorphin levels were decreased in amygdala, spinal cord and pituitary. During naloxone-precipitated abstinence β-endorphin levels were increased in corpus striatum, midbrain and cortex. In addition, in naloxone-precipitated abstinence β-endorphin levels were decreased in pituitary gland and hippocampus but increased in plasma. The levels of methionine-enkephalin in brain regions in decreasing order were: corpus striatum, pons and medulla, amygdala, hypothalamus, midbrain, hippocampus and cortex. The levels of methionine-enkephalin in pons and medulla, amygdala, hippocampus and pituitary gland were decreased in morphine tolerant-dependent rats. During protracted abstinence from morphine, methionine-enkephalin levels in spinal cord, amygdala, pons and medulla, midbrain, cortex, corpus striatum and pituitary gland were decreased. The levels of methionine-enkephalin in hypothalamus and corpus striatum of naloxone-precipitated abstinent rats were increased but were decreased in amygdala and pituitary gland. These results suggest that during morphine tolerance-dependence and during protracted abstinence β-endorphin and methionine-enkephalin levels in discrete brain regions and pituitary gland are decreased. During precipitated abstinence β-endorphin levels are increased in brain regions (except hippocampus) and plasma but decreased in pituitary, whereas methionine-enkephalin levels in amygdala and pituitary gland are decreased except in corpus striatum and hypothalamus where they are increased. The pituitary levels of β-endorphin where reduced in all three conditions. However, the levels after withdrawal were not significantly different from those in tolerant—dependent animals.     

Regional brain concentrations of vasoactive intestinal polypeptide in normotensive Wistar-Kyoto and spontaneously hypertensive rats

The regional brain and spinal cord concentrations of vasoactive intestinal polypeptide immunoreactivity (VIP) were measured in age-matched normotensive Wistar-Kyoto (WKY) and spontaneously hypertensive (SH) rats. The relative order of distribution of VIP in the WKY strain was cortex (44 pmol/g) greater than hippocampus = striatum greater than midbrain = hypothalamus greater than medulla oblongata/pons = lumbar spinal cord (SC) greater than cervical SC greater than thoracic SC (2.5 pmol/g) whereas in the SH strain this order was cortex (35 pmol/g) greater than striatum = midbrain greater than hippocampus = hypothalamus greater than medulla oblongata/pons = lumbar SC greater than cervical SC greater than thoracic SC (1 pmol/g). The VIP concentrations of the thalamus, cerebellum and pituitary were at the level of assay sensitivity (0.5 pmol/g) in both strains. In comparison to the WKY, the SH rats had significantly lower VIP levels in the hippocampus (-42%) and cervical (-46%) and thoracic (-56%) spinal cord but significantly higher levels in the midbrain (+64%).     

褪黑素对母婴分离新生鼠行为发育及海马糖皮质激素受体的调节

目的 探讨外源性褪黑素(MT)对母婴分离(MS)新生大鼠行为发育及海马糖皮质激素受体(GR)表达的调节作用.方法 按随机数字表法将新生同窝雄性SD大鼠随机分为母婴分离(MS)组、母婴分离+褪黑素干预(MT)组和对照组,每组12只.MT组和MS组于出生后第1～14天进行母婴分离实验;MT组于出生后第7～20天腹腔注射MT 10 mg/(kg·d),注射容积10 ml/kg,MS组和对照组分别腹腔注射相同容积的生理盐水.于出生后第28天,采用水迷宫试验测试大鼠学习记忆能力;于出生后第30天,取3组大鼠海马组织,采用二步法免疫组织化学染色检测GR阳性细胞表达,采用逆转录聚合酶链式反应法检测GR mRNA表达.结果 (1)MS组大鼠水迷宫测试学会正确上台所需的训练次数[(13.3±3.7)次],较对照组[(8.9±2.1)次]明显增加(P＜0.01),MT组大鼠学会正确上台所需的训练次数[(9.2±2.6)次],较MT组明显减少(P＜0.01),与对照组比较差异无统计学意义(P＞0.05);(2)MS组海马GR蛋白(78.56±4.05)及mRNA表达(0.65±0.28)较对照组[(136.04±5.16)、(0.95±0.34)]均明显下降,差异有统计学意义,MT组大鼠海马GR蛋白( 129.11±5.03)及mRNA (0.94±0.32)表达明显升高(P＜0.05),与对照组比较差异无统计学意义(P＞0.05).结论 外源性MT可能通过上调海马GR水平减轻早期母婴分离对远期行为发育的损伤.     

经侧脑室注射IL-1ra对脑挫裂伤大鼠血浆S-100β蛋白和神经功能的影响

目的 研究白细胞介素-1受体拮抗剂(IL-1ra)对大鼠脑挫裂伤后血浆S-100β蛋白含量变化和大鼠神经功能行为评分的影响,探讨IL-1ra脑保护作用的可能机制.方法 雄性Wistar大鼠采用随机数字表法分为生理盐水组(n=30)及IL-1ra组(n=30),分别经侧脑室注射生理盐水、IL-1ra 5 μL,并设立正常对照组(n=5).30 min后按Feeney法将生理盐水组及IL-1ra组大鼠制作出脑挫裂伤模型,正常对照组不做任何处理.采用酶联免疫吸附试验(ELISA)测定脑挫裂伤后6 h、12 h、24 h、2 d、3 d、7 d各组大鼠血浆S-100β蛋白含量,Faden评分标准评定大鼠颅脑创伤后各时间点神经功能行为.结果 (1)正常对照组大鼠血浆S-100β蛋白含量为(0.43±0.04)μg/L,神经功能评分为35分.(21伤后各时间点生理盐水组及IL-1ra组大鼠血清S-100β蛋白含量显著高于正常对照组(P<0.05).(3)IL-1ra组各时间点大鼠血清S-100β蛋白含量低于生理盐水组,且差异有统计学意义(P<0.05).(4)IL-1ra组伤后各时间点神经功能评分显著高于生理盐水组(P<0.05).结论 (1)IL-1ra可降低脑挫裂伤大鼠血浆S-100β蛋白含量,提高大鼠神经功能评分.(2)IL-1ra对脑挫裂伤大鼠具有脑保护作用,可能与IL-1ra阻断IL-1β介导的损伤性脑细胞炎症反应有关.     

A single meal elicits regional changes in bombesin-like peptide levels in the gut and brain.

Several studies have demonstrated that exogenous bombesin (BN) elicits a potent satiety effect when administered centrally or systemically. It has been suggested that BN-like peptides may play a physiological role in the control of food intake. The objective of the present study was to determine whether the levels of endogenous BN-like peptides change in response to a meal. All rats were food deprived overnight for a 12 h period. Half the animals were then allowed to feed for 35 min (postprandial group) and the remainder formed the preprandial group. Regions of the brain (hypothalamus, cerebellum, medulla, pons, neocortex, hippocampus, olfactory bulbs, striatum, midbrain and pituitary), gut (oesophagus, fundus, antrum, duodenum, jejunum, ileum, colon) and adrenal glands were analyzed for BN-like peptide levels using radioimmunoassay. Our results indicate significant increases in the levels of BN-like peptides in the hypothalamus and hippocampus, as well as in the antrum of the stomach, after food ingestion. These results are the first to demonstrate the activation of endogenous BN-like peptide mechanisms in response to ingestion. These rapid alterations in peptide levels may support the contention that BN-like peptides play a physiological role in the regulation of ingestive behavior.