次声作用对大鼠海马PSA-NCAM表达的影响

目的研究实验大鼠海马区多聚唾液酸神经细胞黏附因子（PSA-NCAM）经次声作用后的表达情况。 方法将96只SD大鼠随机分为次声作用组和对照组,将次声作用组大鼠暴露于16 Hz,130 dB次声环境中,2 h/d;对照组大鼠于相同时间置于次声压力舱内,但期间不给予次声干预。采用免疫组织化学染色法观察次声作用后不同时间点（次声作用后第1、7、14及21天）及次声结束后即日、第7天和第14天时大鼠海马区PSA-NCAM的表达情况。 结果大鼠海马区PSA-NCAM于次声作用1 d后即开始增加,于次声连续作用第14天时达到峰值,第21天后有所降低,但仍显著高于对照组水平（P<0.05）。当停止次声作用后,发现实验大鼠海马区PSA-NCAM表达水平逐渐下降,于第14天后达到最低值,但仍显著高于对照组水平（P<0.05）。 结论16 Hz 130 dB次声作用可引起大鼠海马区PSA-NCAM表达增加;当停止次声作用后,实验大鼠海马区PSA-NCAM表达随时间延长而逐渐恢复,提示次声所致脑损伤能促进神经干细胞迁移,可能与受损神经修复有关。     

吗啡依赖性大鼠海马CA4区神经细胞的改变

目的：观察吗啡依赖性大鼠海马CA4区神经激肽B及神经肽Y细胞的变化，探讨其发病机制。方法：用皮下注射吗啡法建立雄性大鼠吗啡依赖模型。用免疫组织化学和图像分析方法观察大鼠CA4区神经激肽B和神经肽Y细胞的变化。结果：正常对照组大鼠海马CA4区单位面积神经激肽B，神经肽Y阳性反应物平均灰度值为98．3&;#177;6．3，197．5&;#177;6．14，实验组为215．2&;#177;9．1，238．4&;#177;8．35，与正常对照组比较，实验组神经激肽B，神经肽Y阳性反应物大量减少，差异有显著性意义(P&;lt;0．01)。结论：神经激肽B及神经肽Y细胞减少与吗啡依赖性的发生、发展有关。     

吗啡依赖性大鼠海马CA4区神经细胞的改变

目的:观察吗啡依赖性大鼠海马CA4区神经激肽B及神经肽Y细胞的变化,探讨其发病机制。方法:用皮下注射吗啡法建立雄性大鼠吗啡依赖模型。用免疫组织化学和图像分析方法观察大鼠CA4区神经激肽B和神经肽Y细胞的变化。结果:正常对照组大鼠海马CA4区单位面积神经激肽B,神经肽Y阳性反应物平均灰度值为98.3±6.3,197.5±6.14,实验组为215.2±9.1,238.4±8.35,与正常对照组比较,实验组神经激肽B,神经肽Y阳性反应物大量减少,差异有显著性意义(P<0.01)。结论:神经激肽B及神经肽Y细胞减少与吗啡依赖性的发生、发展有关。     

人骨髓间充质干细胞移植促进脑梗死大鼠神经细胞再生和神经功能恢复

背景：有研究表明,骨髓间充质干细胞或神经干细胞同种移植可改善脑梗死模型鼠神经功能。目的：观察人骨髓间充质干细胞在移植大鼠脑神经功能梗死局部的内存活、迁移情况及神经细胞再生。方法：将人骨髓间充质干细胞经静脉注入免疫抑制的大脑中动脉闭塞模型大鼠,每周观察、记录移植大鼠的行为和神经功能状况（NSS评分、转棒实验）;于移植后1~8周处死大鼠,采用免疫组织化学染色,免疫荧光染色和RT-PCR检测检测移植大鼠大脑人细胞核抗原、神经干细胞标志物（巢蛋白）及神经细胞标志物（神经元核心抗原、脑型微管蛋白、胶质纤维酸性蛋白）的表达。结果与结论：骨髓间充质干细胞移植后2周,脑梗死模型鼠NSS评分明显降低（P〈0.05）、转棒实验停留时间明显延长（P〈0.01）,神经功能得到改善（P〈0.05）。人骨髓间充质干细胞移植后1~8周,在移植大鼠病变侧室管膜下区、海马齿状回、梗死灶周围均可检测到人细胞核抗原阳性细胞。移植后一二周,在移植大鼠病变侧室管膜下区、海马齿状回（CA1,CA3区）、梗死灶周围可见部分巢蛋白阳性细胞和mRNA表达,存续至8周。移植后2周,在移植大鼠病变侧室管膜下区、海马齿状回、梗死灶周围可见部分神经元核心抗原、脑型微管蛋白阳性细胞和mRNA的表达。移植后4~8周,在海马、梗死灶周围可见少量胶质纤维酸性蛋白阳性细胞和少量mRNA的表达。结果证实,静脉移植人骨髓间充质干细胞能在脑梗死模型大鼠脑内存活、并迁移至病变部位,继而分化为神经细胞,促进新生神经细胞的形成,改善移植大鼠的神经功能。     

人骨髓间充质干细胞移植促进脑梗死大鼠神经细胞再生和神经功能恢复

背景:有研究表明,骨髓间充质干细胞或神经干细胞同种移植可改善脑梗死模型鼠神经功能.目的:观察人骨髓间充质干细胞在移植大鼠脑神经功能梗死局部的内存活、迁移情况及神经细胞再生.方法:将人骨髓间充质干细胞经静脉注入免疫抑制的大脑中动脉闭塞模型大鼠,每周观察、记录移植大鼠的行为和神经功能状况(NSS评分、转棒实验);于移植后1~8周处死大鼠,采用免疫组织化学染色,免疫荧光染色和RT-PCR检测检测移植大鼠大脑人细胞核抗原、神经干细胞标志物(巢蛋白)及神经细胞标志物(神经元核心抗原、脑型微管蛋白、胶质纤维酸性蛋白)的表达.结果与结论:骨髓间充质干细胞移植后2周,脑梗死模型鼠NSS评分明显降低(P < 0.05)、转棒实验停留时间明显延长(P < 0.01),神经功能得到改善(P < 0.05).人骨髓间充质干细胞移植后1~8周,在移植大鼠病变侧室管膜下区、海马齿状回、梗死灶周围均可检测到人细胞核抗原阳性细胞.移植后一二周,在移植大鼠病变侧室管膜下区、海马齿状回(CA1,CA3区)、梗死灶周围可见部分巢蛋白阳性细胞和mRNA表达,存续至8周.移植后2周,在移植大鼠病变侧室管膜下区、海马齿状回、梗死灶周围可见部分神经元核心抗原、脑型微管蛋白阳性细胞和mRNA的表达.移植后4~8周,在海马、梗死灶周围可见少量胶质纤维酸性蛋白阳性细胞和少量mRNA的表达.结果证实,静脉移植人骨髓间充质干细胞能在脑梗死模型大鼠脑内存活、并迁移至病变部位,继而分化为神经细胞,促进新生神经细胞的形成,改善移植大鼠的神经功能.     

电针调控海马头蛋白mRNA的表达对慢性脑低灌注大鼠学习记忆及海马神经发生的影响

目的:通过电针刺激慢性脑低灌注模型大鼠百会穴和大椎穴,观察海马组织头蛋白(Noggin)mRNA的表达,以及电针对慢性脑低灌注模型大鼠学习记忆和海马神经发生的影响。方法:选取雄性Sprague Dawley(SD)大鼠120只,采用改良永久性结扎双侧颈总动脉法制作慢性脑低灌注模型,将造模成功的104只大鼠分为模型组和电...     

人胚神经干细胞植入大鼠脑梗死区后的存活和分化

背景:神经干细胞移植可明显改善受损的神经功能,但在体内外多种因素的影响下,移植的神经干细胞其分化数量和分化方向受到限制.目的:观察人胚神经干细胞植入大鼠脑梗死区后的存活、迁移和分化情况.设计、时间及地点:细胞学体内观察,于2007-04/2008-04在泸州医学院附属医院分子生物实验室完成.材料:SD雌性大鼠30只,由中国医学科学院实验动物研究提供.流产胎儿由泸州医学院附属医院妇产科提供.方法:取流产胎儿大脑皮质,剪碎后消化离心,过滤后取沉淀细胞重新悬浮,加入含成纤维细胞生长因子、表皮细胞生长因子、白血病抑制冈子的DMEM/F12培养基,体外培养获得人胚神经干细胞.30只大鼠通过结扎颈外动脉建立脑梗死模型,行走时向右侧转圈者为成功模型,剔除5只无上述明显症状的失败模型鼠.于造模后24 h,以前囟尾侧0.8 mm,中线右外侧1.2 mm为注射点,每只大鼠移植行BrdU标记的1×109 L-1人胚神经干细胞悬液10 μL,深度为硬脑膜下3.0～3.2 mm.主要观察指标:免疫组织化学及免疫荧光染色观察植入的人胚神经干细胞在脑梗死区的存活、迁移和分化状态.结果:人胚神经干细胞体外培养48 h后聚集成球悬浮生长,传三四代后加入血清诱导可见巢蛋白免疫荧光染色呈阳性,发出绿色荧光并聚集成球.BrdU阳性细胞为椭圆形棕褐色,移植后第2,4 周均可见其在脑梗化区存活片向周围迁移,且第4周时迁移的范围更广.移植后2周,皮质颗粒层和皮质下均见较多的BrdU阳性细胞,目BrdU/微管相关蛋白2双阳性细胞多于BrdU/胶质纤维酸件蛋白双阳性细胞;移植后4周脑梗死区BrdU阳性细胞数明显减少,主要存在于脉络丛和微血管中,且BrdU/胶质纤维酸性蛋白双阳性细胞多于BrdU/微管相关蛋白2双阳性细胞.结论:人胚神经干细胞能存活于脑梗死的环境中,并逐渐分化成神经元或星形胶质细胞.移植后期脑实质内存活的神经干细胞明显减少,大量迁移到侧脑室脉络丛和脑表面微血管内,被内皮吞噬系统消化.     

大鼠骨髓基质干细胞诱导分化神经细胞的实验研究

目的探讨骨髓源性神经干细胞分化为神经细胞的可行性，为中枢神经组织损伤修复寻找种子细胞的理想来源。方法无菌取成熟大鼠长骨骨髓，密度梯度离心分离后获得骨髓基质细胞，在神经干细胞培养液及细胞因子等条件下诱导其分化为神经元样细胞，通过免疫细胞化学方法对诱导后细胞进行鉴定。结果骨髓基质细胞能在神经干细胞条件培养基中增殖、分化，表达巢蛋白抗体抗原，在适宜诱导分化因子及条件下进一步分化为神经元样细胞，免疫细胞化学检测可见有神经细胞特异性核蛋白抗体抗原表达阳性。结论在适宜培养条件及诱导因子联合作用下，骨髓基质细胞可成功诱导出神经元样细胞。     

中药干预对创伤性应激障碍大鼠海马区移植间充质干细胞的保护作用

背景：骨髓间充质干细胞移植修复损伤海马具有较高的研究潜力,但是目前移植后干细胞的成活率较低这一问题尚未得到有效解决。目的：综述中药干预对创伤性应激障碍大鼠海马区移植间充质干细胞的保护作用。方法：应用计算机检索2000年1月至2012年1月PubMed数据库相关文章,检索词为＂mesenchymal stem cells,transplantation,survival,post-traumatic stress disorder（PTSD）,hippocampus＂,并限定文章语言种类为English,同时检索2000年1月至2012年1月CNKI数据库相关文章,检索词为＂间充质干细胞,移植,存活率,创伤后应激障碍,海马＂,并限定文章语言种类为中文;共检索文献109篇。最终纳入符合标准的文献23篇。结果与结论：创伤后应激障碍作为一种由生物、心理、社会等多种因素作用的复杂疾病,治疗是亟待解决的问题,中医药在此领域具有独特的优势。创伤后应激障碍所造成的海马损伤可以通过骨髓间充质干细胞移植进行修复,运用调肝方药逍遥散干预,预期可有效提高移植干细胞的存活率。     

中药干预对创伤性应激障碍大鼠海马区移植间充质干细胞的保护作用

背景:骨髓间充质干细胞移植修复损伤海马具有较高的研究潜力,但是目前移植后干细胞的成活率较低这一问题尚未得到有效解决。目的:综述中药干预对创伤性应激障碍大鼠海马区移植间充质干细胞的保护作用。方法:应用计算机检索2000年1月至2012年1月PubMed数据库相关文章,检索词为"mesenchymal stem cells,transplantation,survival,post-traumatic stress disorder(PTSD),hippocampus",并限定文章语言种类为English,同时检索2000年1月至2012年1月CNKI数据库相关文章,检索词为"间充质干细胞,移植,存活率,创伤后应激障碍,海马",并限定文章语言种类为中文;共检索文献109篇。最终纳入符合标准的文献23篇。结果与结论:创伤后应激障碍作为一种由生物、心理、社会等多种因素作用的复杂疾病,治疗是亟待解决的问题,中医药在此领域具有独特的优势。创伤后应激障碍所造成的海马损伤可以通过骨髓间充质干细胞移植进行修复,运用调肝方药逍遥散干预,预期可有效提高移植干细胞的存活率。     

Cannabinoids promote embryonic and adult hippocampus neurogenesis and produce anxiolytic- and antidepressant-like effects

The hippocampal dentate gyrus in the adult mammalian brain contains neural stem/progenitor cells (NS/PCs) capable of generating new neurons, i.e., neurogenesis. Most drugs of abuse examined to date decrease adult hippocampal neurogenesis, but the effects of cannabis (marijuana or cannabinoids) on hippocampal neurogenesis remain unknown. This study aimed at investigating the potential regulatory capacity of the potent synthetic cannabinoid HU210 on hippocampal neurogenesis and its possible correlation with behavioral change. We show that both embryonic and adult rat hippocampal NS/PCs are immunoreactive for CB1 cannabinoid receptors, indicating that cannabinoids could act on CB1 receptors to regulate neurogenesis. This hypothesis is supported by further findings that HU210 promotes proliferation, but not differentiation, of cultured embryonic hippocampal NS/PCs likely via a sequential activation of CB1 receptors, G(i/o) proteins, and ERK signaling. Chronic, but not acute, HU210 treatment promoted neurogenesis in the hippocampal dentate gyrus of adult rats and exerted anxiolytic- and antidepressant-like effects. X-irradiation of the hippocampus blocked both the neurogenic and behavioral effects of chronic HU210 treatment, suggesting that chronic HU210 treatment produces anxiolytic- and antidepressant-like effects likely via promotion of hippocampal neurogenesis.     

Thyrotropin-releasing hormone: neurogenesis of actions in the pentobarbital narcotized rat

Possible neuroanatomical substrates mediating some of the effects seen with thyrotropin-releasing hormone (TRH) in the pentobarbital (PB) narcotized rat were examined. This was accomplished by microinjecting picomole concentrations of TRH into 20 different brain sites. The behavioural effects examined were the capacity of TRH to antagonize PB-induced narcosis and hypothermia as well as TRH-induced shaking behavior. Microinjection of TRH into the septum was found to be significantly more effective in the reversal of PB narcosis than any other site examined. In contrast, the temperature and shaking response were evoked with approximately equal efficacy by TRH microinjection into a number of brain sites; including the preoptic/anterior hypothalamus, medial thalamus, thalamic, periventricular gray, interpeduncular nucleus and locus ceruleus. These results demonstrate the septal region to be the site of action for TRH reversal of PB narcosis and suggest the involvement of the septohippocampal system. In addition, they indicate that the neurogenesis of the shaking response is similar to that of the temperature response and that this differs from the neurogenesis of the analeptic response.     

成年大鼠弥漫性脑损伤后海马齿状回神经元再生动态变化的实验研究

目的观察弥漫性脑损伤后大鼠海马齿状回神经前体细胞的增殖规律。方法使用BrdU标记分裂细胞、免疫组织化学方法和激光共聚焦显微镜观察弥漫性脑损伤后大鼠海马齿状回神经元再生水平的变化规律。结果成年大鼠弥漫性脑损伤后4～12d时间段内,齿状回BrdU免疫阳性细胞数量表现出明显的统计学差异(P<0.01),其中第6天时达到峰值。这些BrdU阳性细胞大部分分化为神经元。结论弥漫性脑损伤可诱导成年脑海马齿状回细胞再生水平在一定时间内上调。     

Opioid-mediated facilitation of long-term depression in rat hippocampus

Previous studies have demonstrated that opioid substances are often inhibitors of the gamma-aminobutyric acid (GABA) transmitter system in the hippocampal formation, and that GABA-mediated inhibition is a potent modulator of synaptic plasticity. Field excitatory postsynaptic potentials were recorded from the CA1 region of rat hippocampal slices in response to stimulation of the Schaffer collateral fibers to monitor the effects of acute opioid exposure on the induction of long-term depression (LTD) at excitatory synapses in the stratum radiatum. Exogenous application of a selective mu-opioid agonist resulted in a greater than 2-fold enhancement of LTD, whereas kappa- and delta-agonists did not significantly affect LTD magnitude. Costimulation of the opioid peptide-containing stratum lacunosum-moleculare during LTD induction also resulted in a facilitation of LTD in the stratum radiatum, an effect prevented by prior administration of an opioid antagonist. These results suggest that both exogenously applied and endogenously released opioids can act to facilitate LTD of the Schaffer collateral input to CA1 pyramidal neurons.     

β-淀粉样肽对小鼠海马神经发生的影响及中药的干预作用

目的观察Aβ2 5 35对痴呆小鼠模型海马神经发生的影响及中药“脑复聪”的干预作用。方法脑室内注射Aβ2 5 35造成痴呆模型小鼠。用Morris水迷宫测空间学习记忆功能 ;同位素配基测受体结合活性 ;BrdU标记的免疫组化检测神经发生。结果模型小鼠的学习记忆功能降低 (P <0 0 5 ) ;海马NMDA受体结合活性显著升高而胆碱能M受体结合活性显著降低 (P <0 0 0 1) ;海马齿状回BrdU阳性细胞减少 (P <0 0 5 )。经“脑复聪”治疗后上述变化均得到改善 (P <0 0 5 ,P <0 0 0 1,P <0 0 5 )。结论Aβ2 5 35的神经毒机理包括对神经发生的抑制作用 ;提高神经发生率可能是中药治疗神经退行性疾病的重要机理之一。     

Kynurenic acid and quinolinic acid act at N-methyl-D-aspartate receptors in the rat hippocampus

Responses evoked by several amino acid excitants, including the tryptophan metabolite quinolinic acid, were recorded intracellularly from CA1 pyramidal neurons in rat hippocampal slices. Quinolinate, N-methyl-D-aspartate (NMDA), ibotenate and (+/-)-cis-1-amino-1,3-dicarboxycyclopentane produced excitations characterized by burst firing of action potentials, tetrodotoxin-resistant spiking and apparent increases in input resistance measured with brief hyperpolarizing current pulses. L-Glutamate, kainate, quisqualate and (+/-)-2'-amino-3-hydroxy-5-methyl-4-isoxazole-3'-propionate depolarized CA1 pyramidal neurons and induced apparent decreases in input resistance. Quinolinate-, NMDA-, and ibotenate-induced focal depolarizations, but not L-glutamate, kainate- or quisqualate-induced responses, were strongly antagonized by specific NMDA receptor antagonists. The tryptophan metabolite kynurenic acid, at concentrations that antagonized focal depolarizations produced by NMDA, ibotenate and the endogenous excitant quinolinate, did not antagonize quisqualate or L-glutamate responses. In addition to its NMDA-type antagonist action, kynurenate blocked kainate-induced focal depolarizations.     

Effects of chronic administration of adipokinetic and hypertrehalosemic hormone on animal behavior,BDNF, and CREB expression in the hippocampus and neurogenesis in mice

Neurosecretory cells in corpus cardiacum of insects synthesize a set of hormones that are called adipokinetic, hypertrehalosaemic or hyperprolinaemic, depending on insect in question. This study investigated effects of chronic administration of Anax imperator adipokinetic hormone (Ani‐AKH), Libellula auripennis adipokinetic hormone (Lia‐AKH), and Phormia‐Terra hypertrehalosaemic hormone (Pht‐HrTH) on depression, anxiety, analgesy, locomotion in forced swimming (FST), elevated plus‐maze (EPM), hot plate, and locomotor activity tests. Ani‐AKH (1 and 2 mg/kg), Lia‐AKH (1 and 2 mg/kg), and Pht‐HrTH (1 and 2 mg/kg) had antidepressant effects in forced swimming test. Lia‐AKH (2 mg/kg) and Pht‐HrTH (1 and 2 mg/kg) had anxiolytic effects when given chronically in elevated plus‐maze test. Ani‐AKH (1 and 2 mg/kg) and Pht‐HrTH (2 mg/kg) had antinociceptive effects in hot plate test in male balb‐c mice. Ani‐AKH (2 mg/kg), Lia‐AKH (1 and 2 mg/kg), and Pht‐HrTH had locomotion‐enhancing effects in locomotor activity test in male balb‐c mice. Drug treatment significantly increased brain‐derived neurotrophic factor (BDNF) and cyclic adenosine monophosphate (cAMP) response element binding protein (CREB) gene expression levels compared to control levels. Pht‐HrTH and Ani‐AKH groups had significantly increased numbers of BrdU‐labeled cells, while neurodegeneration was lower in the Pht‐HrTH group. Our study showed that AKH/RPCH family peptides may be used in treatment of psychiatric illness such as depression and anxiety, in treatment of pain and in diseases related to locomotion system. AKH/RPCH family peptides increase neurotrophic factors in brain and have potential proliferative and neuroprotective effects in hippocampal neurogenesis and neurodegeneration.     

Eltoprazine suppresses hyperpolarizing responses to serotonin in rat hippocampus

In this study we report the effects of eltoprazine, a phenylpiperazine derivative with high affinity for 5-hydroxytryptamine1 (5HT1) binding sites, on membrane properties of hippocampal neurons. Intracellular recordings were made from cornus ammoni-1 pyramidal neurons in rat hippocampal slices. Responses to eltoprazine were compared with 5HT-induced responses. Superfusion with 5HT induced a dose-dependent hyperpolarization of the membrane accompanied by a resistance decrease. Eltoprazine evoked membrane changes that were similar to but much weaker than those induced by 5HT. Both the 5HT- and eltoprazine-evoked membrane hyperpolarizations were largely suppressed in the presence of spiperone. The eltoprazine-induced effects persisted in the presence of tetrodotoxin and tetraethylammonium and also when haloperidol and phentolamine were added to the medium, indicating that the small agonistic effects of eltoprazine are not due to an indirect activation of dopamine or alpha adrenergic receptors. Superfusion with eltoprazine furthermore resulted in a marked reduction of the response to concomitantly applied 5HT. Dose-response curves for 5HT were shifted to the right in the presence of eltoprazine, while the maximal response was diminished. Hyperpolarizations induced by baclofen, which presumably activates the same K+ conductance as 5HT, were not significantly reduced by eltoprazine. Our data, added to the previously demonstrated high affinity of eltoprazine for 5HT1 sites, suggest that in the hippocampal cornus ammoni-1 area eltoprazine acts as a partial 5HT1 agonist with a relatively low intrinsic activity but a considerable potency to suppress hyperpolarizing responses to 5HT.