易卒中型肾血管性高血压大鼠蓝班NE能神经元系统的研究

用免疫组化(ABC)方法,对易卒中型肾血管性高血压大鼠高血压早期,中期和晚期蓝斑去甲肾上腺素(NE)能神经元系统进行了统计分析.高血压中期手术组和假手术组蓝斑NE能神经元数相似,高血压早期和晚期手术组的稍多于假手术组的,但差异无统计学意义.易卒中型肾血性高血压的各期蓝斑的NE能神经元数目没有多大变化.     

心理应激和体力运动对大鼠海马NE及其合成酶TH、代谢酶MAOA的影响

目的： 本研究通过比较慢性心理应激（以下简称应激）和长期体力运动（以下简称运动）2种过程对大鼠海马去甲肾上腺素（NE)水平及其合成酶酪氨酸羟化酶（TH）和代谢酶单氨氧化酶A（MAOA）的影响,探讨体力运动减缓海马应激损伤的相关机制。方法：动物分别施予自愿跑轮动物模型4周和慢性束缚应激模型3周后,高效液相色谱法比较海马NE变化;蛋白免疫印迹法检测NE合成酶TH、代谢酶MAOA蛋白表达变化。结果：与对照组相比,应激大鼠体重增长缓慢,糖水偏嗜量明显降低,NE水平明显降低［（303.46±24.36) ng/g,P<0.05］,TH、MAOA蛋白表达升高,分别为(1.35±0.94）倍和（2.22±0.62)倍（P<0.05）;运动组大鼠NE水平明显升高｛（418.53±70.64) ng/g,P<0.05］,TH、MAOA蛋白表达亦升高,分别为(1.22±0.59)倍和(1.85±0.36)倍（P<0.05）。结论：结果提示NE水平变化可能与运动减缓应激性海马损伤的关系密切,NE水平的改变与NE的合成与代谢关系不密切,可能存在其它机制。     

易卒中型肾血管性高血压大鼠蓝班NE能神经元系统的研究

用免疫组化方法，对易卒中型肾血管性高血压大鼠高血压早期，中期和晚期蓝斑去甲肾上腺素能神经元系统进行了统计分析。高血压中期手术组和假手术组蓝斑ＮＥ能神经元数相似，高血压早期和晚期手术组的稍多于假手术组的，但差异无统计学意义。易卒中型肾血性高血压的各期蓝斑的ＮＥ能神经元数目没有多大变化。     

内脏高敏感大鼠肠道和脊髓5-羟色胺能神经元及神经纤维分布

目的　研究内脏高敏感大鼠肠道及脊髓内的 5 羟色胺 (5 HT)能神经元及神经纤维的分布 ,肥大细胞脱颗粒对 5 HT能神经元及神经纤维的影响。方法　内脏高敏感大鼠分为两组 (A组和B组 ) ,B组为事先给予促肥大细胞脱颗粒释放剂com pound 4 8 80的内脏高敏感大鼠。对内脏高敏感大鼠和正常对照组 (C组 )结肠和脊髓进行 5 HT免疫组化IHN Envision染色法并作定量分析。结果　A组和C组比较肠道 5 HT免疫阳性神经纤维在黏膜下层阳性指数 (PI)明显增加 (P <0 0 1,0 6 34± 0 2 75vs 0 2 4 5± 0 131) ,肌间神经丛内和脊髓后角 5 H…     

易卒中型肾血管性高血压及高脂血症大鼠脑血管壁NE、NPY能神经纤维的研究

<正>交感神经递质去甲肾上腺素(NE),神经肽Y(NPY)与高血压、动脉粥样硬化及脑卒中等疾病关系密切.实验用健康雄性SD大鼠96只,体重80—110g/只,随机分四组:1.对照组(24只):行假手术、普通饮食;2.高血压组(24只):用双肾双夹法复制易卒中型肾血管性高血压大鼠(RHRSP)模型、普通饮食,3.高脂组(24只):行假手术、高脂饮食;4.高血压+高脂组(24只):复制RHRSP模型、高脂饮食.各组分别手术后1周、4周、12周处死,取双侧大脑中动脉、基底动脉、ABC免疫组化染色,定量分析血管壁NE、NPY阳性神经纤维密集变化,结果发现:1.NE能神经纤维:对照组各期无明显变化;高血压组晚期显著减少(P<0.01);高脂组早     

多重脑震荡大鼠5-羟色胺能神经纤维变化

目的:研究多重脑震荡(MCC)与大鼠认知记忆相关脑区5-羟色胺(5-HT)能神经纤维变化的规律,以探讨MCC大鼠认知行为障碍的神经生物学基础。方法:应用自制单摆式机械打击装置构建MCC大鼠模型,免疫组织化学及半定量方法研究MCC后大鼠5-HT能神经纤维的变化。结果:前额叶皮质、中隔区、丘脑、海马CA1和CA3区5-HT能神经纤维免疫反应阳性物在伤后2d组反应达高峰,杏仁体、海马CA2和CA4区在伤后4d组免疫反应了阳性最强;顶叶皮质、梨状皮质及伏隔核内侧在伤后8d组反应最强。5-HT纤维密度在中隔区和伏隔核内侧以伤后2d最高,杏仁体区以伤后1d最高,大脑皮质和丘脑5-HT纤维密度在伤后各损伤组与对照组相比无显著性差异。结论:MCC可引起与认知功能有关脑区的5-HT表达增高,5-HT的高表达可能由脑损伤后神经元过度兴奋所引起的神经递质紊乱造成,而这种紊乱可能影响大鼠的认知功能。     

N-乙酰半胱氨酸对慢性间歇缺氧大鼠海马神经元凋亡及氧化应激的影响

目的：探讨N-乙酰半胱氨酸(NAC)对慢性间歇缺氧(CIH)模型大鼠海马组织氧化应激及海马神经元凋亡的影响。方法:30只雄性Wistar大鼠随机分为慢性缺氧组、NAC治疗组及正常对照组3组，每组10只。采用化学比色法测定海马组织丙二醛(MDA)、超氧化物歧化酶(SOD)水平，同时应用免疫组化方法检测海马CA1 区磷酸化JNK(p-JNK)表达水平，应用TUNEL法检测海马CA1区神经元凋亡率。结果:NAC治疗组 MDA水平低于CIH组（1.71±0.43 vs 1.37±0.26，P＜0.05）、SOD活性高于CIH组（44.94±14.01 vs 57.66±14.07，P＜0.05），p-JNK表达水平低于CIH组 （0.53±0.10 vs 0.39±0.16，P＜0.05），海马神经元凋亡率显著低于CIH组（0.32±0.18 vs 0.20±0.11，P＜0.05）。结论:NAC能抑制慢性间歇缺氧导致的氧化应激，从而影响JNK信号转导通路，减少海马神经元凋亡。     

不同成熟期大鼠戊四氮反复惊厥模型与癫痫发病机制研究

目的：观察新生大鼠、成熟大鼠反复惊厥后海马结构的变化及NF-κB 表达，探索未成熟脑癫痫发病机制。 方法： 对生后10 d、60 d（P10、P60）两实验组大鼠用戊四氮(PTZ)反复腹腔注射5 d，设P10、P60生理盐水对照组；硫堇染色对CA1、CA3、DG及门区神经元进行细胞计数, 观察海马神经元坏死及凋亡；免疫组化技术检测NF-κB的表达；Timm组织化学染色观察苔藓纤维发芽并评分。 结果： (1) P10实验组与对照组比较，海马齿状回、CA1、CA3区无明显神经元丢失，DG区颗粒细胞数在实验组(23.25±3.06) 多于对照组（16.25±1.58）；P60实验组CA1、CA3区神经元计数（8.22±1.88、5.62±1.68）明显少于对照组（6.31±1.50、3.62±1.40），DG区无明显差异；（2）两实验组CA3区锥体层苔藓纤维发芽均多于对照组，但P60(3.25±1.03)较P10（1.50±0.92）更明显；（3）P10、P60实验组NF-κB 表达高于对照组，且P10组较P60组更为明显（P<0.05）。 结论： 新生大鼠致痫后海马神经元无明显丢失，脑内NF-κB 表达增加，可能是未成熟脑对惊厥性脑损伤具有耐受性的重要神经生物学基础。     

大鼠多重脑震荡后多巴胺能与胆碱能神经元的变化

目的:观察大鼠多重脑震荡(MCC)后中枢神经系统内多巴胺能神经纤维与胆碱能神经元的变化。方法:用金属单摆打击装置复制MCC大鼠模型,随机分为对照组和伤后1、2、4、8、16、24 d多个损伤组,用免疫组织化学双标技术检测大鼠脑内多巴胺能神经纤维中酪氨酸羟化酶(TH)和胆碱能神经元中胆碱乙酰转移酶(ChAT)的表达变化。结果:TH阳性表达在伤后1 d开始上升,8 d达高峰,24 d组基本恢复;ChAT阳性表达在MCC后均呈现下调改变。结论:多重脑震荡后多巴胺能神经纤维和胆碱能神经元的变化可能与中枢神经系统的认知功能障碍有关。     

大鼠蓝斑内P物质、神经降压肽和生长抑素样神经元的上行投射

本实验采用HRP逆行追踪与免疫细胞化学相结合的方法,对大鼠蓝斑内P物质(SP)、神经降压肽(NT)、生长抑素(SOM)三种肽能投射神经元的纤维联系及其局部关系进行了探讨。结果证实:1.HRP注射至丘脑或尾壳核后,大鼠蓝斑内均可见到双标记神经元。2.存在SP样和NT样蓝斑—丘脑纤维,前者终止于丘脑腹后核和板内核,后者终止于丘脑板内核。3.蓝斑至尾壳核的投射呈SOM样免疫反应。4.蓝斑内三种肽能投射的起始和终止部位有所不同,其中SP样和NT样神经元主要位于蓝斑后部背侧区,其纤维终于丘脑;而SOM样投射神经元则见于蓝斑前部背侧区,纤维终止于尾壳核。上述肽能通路均为双侧性,而以同侧占优势。5.蓝斑内三种肽能单标记神经元中以NT样免疫反应阳性胞体最为丰富。以上结果表明;蓝斑上行性投射中除含去甲肾上腺素纤维外,还含有SP样、NT样和SOM样纤维,并存在一定的局部定位投射关系。     

Conditions for adrenergic hyperinnervation in hippocampus: II. Electrophysiological evidence from intraocular double grafts

Summary Following sequential intraocular transplantations of areas containing NE cell bodies (locus coeruleus or superior cervical ganglion) and of NE fiber target areas (hippocampus), both pieces mature in a manner analogous to that observed for individual transplants. NE-containing nerve fibers, derived from either LC or SCG transplants, can be seen to invade the hippocampal formation. When LC is used, the invading fibers markedly hyperinnervate the hippocampus while SCG-derived fiber densities approximate those seen with innervation from the adrenergic ground plexus of the iris. Electrophysiological recordings from neurons in the LC reveal an atropine-sensitive excitatory response to illumination, suggesting innervation of the LC by cholinergic nerve fibers from the iris. This is supported by the fact that dense cholinesterase-positive staining can be found in the LC piece. Application of an epileptogenic agent, such as penicillin, results in a marked excitation of neurons in the LC without inducing epileptiform activity in the hippocampus. In contrast, single hippocampal grafts seize readily after penicillin. Local application of the inhibitory agent GABA into the LC allows penicillin-induced epileptiform activity to generate in the hippocampus, suggesting that functional inhibitory innervation develops between NE fibers derived from LC and pyramidal neurons in the hippocampus. Supporting this, subsequent excitation of LC neurons by iontophoresis of glutamate terminates the hippocampal seizure. Prior administration of reserpine (2.5 mg/kg) disrupts the inhibitory influence of LC innervation on the hippocampal EEG following penicillin. After reserpine, the hippocampal portions of double grafts behave like single hippocampal transplants.It is concluded that sequential transplantations of cell body and target regions of the CNS to the anterior chamber of the eye creates a functional, yet isolated, neuronal pathway which can be utilized to study the development of neuronal connections. Offprint requests to: Dr. Barry J. Hoffer, Dept. of Pharmacology, C236, University of Colorado Health Sciences Center, 4200 E. Ninth Avenue, Denver, CO 80262, USA     

A comprehensive analysis of the effect of DSP4 on the locus coeruleus noradrenergic system in the rat

Degeneration of the noradrenergic neurons in the locus coeruleus (LC) is a major component of Alzheimer's (AD) and Parkinson's disease (PD), but the consequence of noradrenergic neuronal loss has different effects on the surviving neurons in the two disorders. Therefore, understanding the consequence of noradrenergic neuronal loss is important in determining the role of this neurotransmitter in these neurodegenerative disorders. The goal of the study was to determine if the neurotoxin N-(2-chloroethyl)-N-ethyl-2-bromobenzylamine (DSP4) could be used as a model for either (or both) AD or PD. Rats were administered DSP4 and sacrificed 3 days 2 weeks and 3 months later. DSP4-treatment resulted in a rapid, though transient reduction in norepinephrine (NE) and NE transporter (NET) in many brain regions receiving variable innervation from the LC. Alpha₁-adrenoreceptors binding site concentrations were unchanged in all brain regions at all three time points. However, an increase in α₂-AR was observed in many different brain regions 2 weeks and 3 months after DSP4. These changes observed in forebrain regions occurred without a loss in LC noradrenergic neurons. Expression of synthesizing enzymes or NET did not change in amount of expression/neuron despite the reduction in NE tissue content and NET binding site concentrations at early time points, suggesting no compensatory response. In addition, DSP4 did not affect basal activity of LC at any time point in anesthetized animals, but 2 weeks after DSP4 there is a significant increase in irregular firing of noradrenergic neurons. These data indicate that DSP4 is not a selective LC noradrenergic neurotoxin, but does affect noradrenergic neuron terminals locally, as evident by the changes in transmitter and markers at terminal regions. However, since DSP4 did not result in a loss of noradrenergic neurons, it is not considered an adequate model for noradrenergic neuronal loss observed in AD and PD.     

Altered Expression of Tyrosine Hydroxylase in the Locus Coeruleus Noradrenergic System in Citalopram Neonatally Exposed Rats and Monoamine Oxidase A Knock Out Mice

In rodents, noradrenergic (NE) locus coeruleus (LC) neurons are well known to express tyrosine hydroxylase (TH) immunoreactivity. However, due to its very low enzyme activity, NE cortical fibers do not typically express TH immunoreactivity, thus dopamine‐β‐hydroxylase (DBH) immunoreactivity is commonly utilized as a marker for NE cortical fibers. In this study, we performed double and/or triple immunofluorescent staining using antibodies against TH, DBH, and/or norepinephrine transporter (NET) to investigate the altered NE TH expression of cortical fibers in citalopram (CTM)‐exposed rats and monoamine oxidase (MAO) A knock out (KO) mice. We have noted the following novel findings: (1) neonatal exposure to the selective serotonin reuptake inhibitor (SSRI) CTM enhanced NE TH immunoreactive fibers throughout the entire neocortex, and a few of them appeared to be hypertrophic; (2) slightly enhanced NE cortical TH immunoreactive fibers were also noted in MAO A KO mice, and many of them revealed varicosities compared with the rather smooth NE cortical TH immunoreactive fibers in wild‐type (WT) mice; (3) LC dendrites of MAO A KO mice exhibited beaded morphology compared with the smooth LC dendrites in WT mice. Our findings suggest that both genetic and environmental factors during early development may play a critical role in the regulation and proper function of NE TH expression in the neocortex. Anat Rec, 2011. © 2011 Wiley‐Liss, Inc.     

实验性糖尿病神经和微血管形态研究及黄腐酸钠早期干预

目的 :观察糖尿病大鼠神经和神经内膜毛细血管形态变化及黄腐酸钠早期干预效果。方法 :腹腔注射链脲佐菌素建立糖尿病大鼠模型。给予黄腐酸钠治疗 6个月。结果 :(1)糖尿病动物血糖明显增高。(2 )糖尿病组有髓神经纤维密度明显低于对照组。黄腐酸钠治疗组有髓神经纤维密度明显高于糖尿病组。(3)电镜下糖尿病组神经纤维、神经内膜毛细血管出现结构改变 ;黄腐酸钠治疗组上述改变较轻。结论 :提示糖尿病大鼠病程 6个月时出现早期糖尿病神经病变及神经内膜毛细血管结构改变 ,黄腐酸钠可一定程度地抑制其进展。     

Age-induced changes in single locus coeruleus brain transplants grown in oculo: an in vivo electrochemical study.

Brain stem tissue from fetal Sprague-Dawley rats containing the nucleus locus coeruleus (LC) was transplanted into the anterior chamber of the eye of young adult host rats and was studied at 4-6 months (young control) or 24-28 months after grafting (old). High-speed in vivo electrochemical measurements were used to characterize the potassium-evoked synaptic overflow of norepinephrine (NE) in both young and aged LC brain grafts. The amplitudes of potassium-evoked NE overflow were attenuated in the aged grafts as compared to the young LC grafts. In addition, the rise times of potassium-evoked responses were longer in the old LC grafts than in the young transplants. In contrast, the NE content of aged LC grafts, as determined by high-performance liquid chromatography coupled with electrochemical detection (HPLC-EC), was only slightly diminished and not significantly different from the NE levels seen in young LC grafts. However, light microscopical evaluation using tyrosine-hydroxylase immunocytochemistry revealed pyknotic cell bodies and fluorescent accumulations in aged locus coeruleus transplants which were indicative of degeneration in these grafts. The present data demonstrate a significant age-related decline in the presynaptic function of NE-containing neurons in intraocular locus coeruleus transplants of Sprague-Dawley rats.     

氟达拉滨联合瘤苗抗肿瘤生物治疗作用的实验研究

目的探讨氟达拉滨对小鼠CD4＋CD25＋Treg细胞的影响，同时研究其抗肿瘤作用。方法氟达拉滨或生理盐水分别腹腔注射10只小鼠，用流式细胞术检测CD4＋CD25＋Treg细胞相对量。氟达拉滨或生理盐水分别腹腔注射10只小鼠，4d后用丝裂霉素灭活的肿瘤细胞免疫小鼠，观察小鼠抗肿瘤的能力（观察肿瘤发生率和出瘤时间）；用乳酸脱氢酶杀伤实验进一步验证氟达拉滨可增强CTL的杀伤活性。结果氟达拉滨组CD4＋CD25＋Treg细胞占淋巴细胞百分比为（1．150±0．256）％，对照组为（1．488±0．270）％，氟达拉滨组明显低于对照组（P〈0．05）；氟达拉滨＋接种瘤苗组、生理盐水＋接种瘤苗组、氟达拉滨＋未接种瘤苗组和生理盐水＋未接种瘤苗组小鼠第13天肿瘤发生率分别为10％、30％、50％和70％，两两间差异均有统计学意义（P〈0．05）；氟达拉滨＋接种瘤苗组、生理盐水＋接种瘤苗组、氟达拉滨＋未接种瘤苗组和生理盐水＋未接种瘤苗组小鼠分别在接种后第13天、第10天、第8天和第5天发现出瘤现象；对照组的肿瘤生长曲线较为陡直，氟达拉滨组的生长曲线较为平缓。氟达拉滨联合瘤苗接种组和单纯瘤苗接种CTL的活性分别为24．425±13．544和17．575±10．260，两者间差异有统计学意义（P〈0．05）。结论低剂量氟达拉滨降低CD4＋CD25＋Treg细胞，使其抗肿瘤免疫作用增强，联合接种灭活瘤苗进一步增强抵抗肿瘤攻击的能力。     

Reduced responsiveness of locus coeruleus neurons to cutaneous thermal stimuli in capsaicin-treated rats

Recent electrophysiological experiments have shown that brain norepinephrine (NE) neurons in the locus coeruleus (LC) are activated by cutaneous thermal stimuli of both non-noxious and noxious character. In the present study the LC neuronal response to thermal stimuli was used to evaluate cutaneous thermal sensitivity in capsaicin-treated rats, a treatment that is described to cause impaired thermoregulation. Capsaicin treatment, of neonates as well as of adult rats, caused a reduced responsiveness of brain LC neurons to thermal stimuli. The results suggest that a reduction in peripheral thermal afferent transmission may be one mechanism underlying the capsaicin-induced thermoregulatory dysfunction.     

Wortmannin通过抑制癫痫大鼠海马自噬活性发挥神经保护作用

目的:探讨癫痫大鼠海马自噬活性的变化及自噬抑制剂wortmannin(WM)对致痫大鼠海马神经元的保护作用。方法:实验大鼠分为对照组、致痫2 h、8 h、16 h、24 h、72 h组和WM干预组。应用HE染色和Nissl染色观察癫痫大鼠海马神经元损伤的变化。免疫印迹检测海马组织微管相关蛋白1轻链3(LC3)Ⅱ/LC3Ⅰ的比值作为自噬活性的指标。结果:LC3Ⅱ/LC3Ⅰ比值在大鼠癫痫发作2 h时开始升高,24 h达到高峰,持续升高至少72 h。致痫24 h时海马CA1区出现明显神经元损伤和丢失。WM干预组CA1区存活神经元数目(100.88±18.73)显著高于癫痫组(70.16±5.09)(P0.05),LC3Ⅱ/LC3Ⅰ的比值低于癫痫组(P0.05)。结论:癫痫发作导致的海马损伤时存在自噬激活现象;WM通过抑制自噬活性减轻癫痫发作所致的海马损伤,具有神经保护作用。     

巨噬细胞培养上清液促进大鼠坐骨神经损伤后修复的机制研究

目的 探讨巨噬细胞培养上清液促进大鼠坐骨神经损伤后修复的效果及机制.方法 SD雄性大鼠30只,体重200～250 g,随机分成对照组、观察组和模型组,每组10只,成功制作大鼠坐骨神经损伤离断模型后,对照组在吻合口间隙注射给予等量生理盐水,观察组注射巨噬细胞培养上清液,缝合后饲养12周处死。结果 观察组与对照组大鼠均无死亡。与对照组比,观察组大鼠坐骨神经电生理检测波幅[(0.16±0.04)V比(0.33±0.05)V,t=7.45,P=3.87E-05]和传导速度[(13.22±6.23)m/s比(24.54±6.36)m/s,t=4.02,P=3.01E-3)]显著提高,潜伏期[(0.74±0.06)ms比(0.53±0.04)ms,t=9.21,P=7.07E-06) ]缩短,差异有统计学意义.观察组坐骨神经形态学分析平均髓鞘厚度[(0.48±0.07)μm比(1.27±0.08)μm,t=23.50,P=2.18E-09) ]?神经纤维数量[(0.69±0.08)/HP比(3.22±0.06)/HP,t=80.01,P=3.77E-14) ]和有髓神经纤维平均直径[(1.08±0.39)μm比(3.63±0.42)μm,t=14.07,P=1.97E-07) ]显著增加,差异有统计学意义(P<0.05).观察组雪旺细胞表达的神经生长因子mRNA(NGF mRNA)[(0.13±0.04)比(0.42±0.05),t=14.32,P=1.68E-07) ]和层粘连蛋白mRNA[(0.07±0.03)比(0.38±0.06),t=14.61,P=1.41E-07) ]含量显著升高,差异有统计学意义(P<0.05).结论 巨噬细胞培养上清液可以有效促进大鼠坐骨神经损伤后修复,可能与促进雪旺细胞表达NGF和Laminin有关.     

Functional reorganization of the noradrenergic system after partial fornix section: a behavioral and autoradiographic study

Previous experiments revealed that the cholinergic deficit in rats with a partial fornix section was accompanied by an increase in turnover of noradrenaline (NE) in the hippocampus. This noradrenergic hyperactivity contributed to the cognitive deficit in lesioned rats, probably by interaction with the cholinergic system. The present experiment examines the reorganization of the noradrenergic system after the damage induced by partial fornix section and attempts to determine if the increase in NE turnover is of locus coeruleus (LC) origin, or if it is a result of local regulation at the noradrenergic terminals. Rats were submitted to knife-cut section of the fornix, resulting in a decrease in choline acetyltransferase activity in the hippocampus, correlated with a significant behavioral deficit in a spatial memory task. Lesioned rats learned a nonspatial memory task normally. Sections of brains of these rats were submitted to quantitative autoradiography. [¹²⁵I]Iodopindolol binding was assessed in the dorsal and ventral hippocampus to determine availability of receptors. This was found to be significantly lower in lesioned rats. [¹²⁵I]Iodoclonidine was used to determine ₂ receptors binding in dorsal and ventral hippocampus and in LC. There was no difference in ₂ receptors in LC, a significant decrease in dorsal regions of the hippocampus, and a significant increase in ventral regions. Muscarinic M1 receptors in the hippocampus showed no changes after the lesion.