活化淋巴细胞及其染色质诱导抗核抗体生成

系统性红斑狼疮 (ＳＬＥ)是自身免疫病的原型 ,其最重要特征是血清中出现以抗双链ＤＮＡ(ｄｓＤＡＮ)、抗Ｓｍ等为主的致病性抗核抗体 (ＡＮＡ)。阐明抗ｄｓＤＮＡ和抗Ｓｍ抗体生成的机制 ,对了解ＳＬＥ发病机制有重要意义。虽然多种假说来解释ＡＮＡ生成的机理 ,但许多证据提示是抗原驱动ＡＮＡ生成。令人困惑不解的是哺乳类动物ｄｓＤＮＡ、组蛋白、核小体等核成分并不具有免疫原性 ,因此启动生成的激发原迄今不明。一、活化的淋巴细胞同系免疫诱导抗核抗体生成1 细菌活化的淋巴细胞 :1 996年 ,我们首次发现经空肠弯曲菌 (ＣＪ- 1 0 1 )…     

NMDA受体拮抗剂MK-801抑制大鼠甲醛炎症性痛过程中 脊髓后角NOS及NO含量的增加

痛及痛过敏的形成 ,与脊髓伤害性信息传递通路中兴奋性氨基酸 (ＥＡＡｓ)的释放及其受体的活动增强以及表达上调有关 ,这种由于ＥＡＡ受体 ,特别是Ｎ -甲基 -Ｄ -天门冬氨酸 (ＮＭＤＡ)受体活动所引起的痛及痛过敏可被ＮＯＳ抑制剂所阻断。此外 ,一些实验结果也表明痛及痛过敏的形成中脊髓后角神经元中一氧化氮合酶 (ＮＯＳ)的表达增多 ,但是 ,这种ＮＯＳ表达增多的机制尚不清楚 ,是否是ＥＡＡｓ及其受体活动的结果有待于证实。目的 :观察ＮＭＤＡ受体拮抗剂ＭＫ - 80 1对甲醛炎症性痛及痛过敏过程中脊髓后角ＮＯＳ及一氧化氮 (ＮＯ)含量变…     

BDNF、NGF对AD模型鼠脑移植区一氧化氮合酶阳性神经元发育生长的影响

用使君子酸（ＱＡ）损毁ＳＤ 大鼠左侧Ｍ ｅｙｎｅｒｔ基底大细胞核,制成老年性痴呆症（ＡＤ）模型。将同种鼠胚基底前脑制成的细胞悬液不加神经营养因子和分别加入脑源性神经营养因子（ＢＤＮＦ）、神经生长因子（ＮＧＦ）及ＢＤＮＦ＋ ＮＧＦ者植入ＡＤ 模型鼠额、顶叶皮质,隔日通过脑室灌注人工脑脊液和相应神经营养因子共７ 次。移植后４ 个月,取脑切片作Ｎｉｓｓｌ染色、ＮＡＤＰＨ ｄ和ＮＡＤＰＨ ｄ＋ ＡＣｈＥ 组化染色,计数移植区中ＮＯＳ阳性神经元数及其纤维在１６９００ μｍ ２ 网格中的交点数,并用ＭＩＡＳ ３００ 计算机图像分析系统对移植区中ＮＯＳ阳性神经元的细胞面积进行处理。结果显示：给予神经营养因子的动物,移植区中的ＮＯＳ阳性神经元数、ＮＯＳ阳性纤维交点数和ＮＯＳ阳性神经元的细胞面积等形态学指标均较不给予神经营养因子的对照组为佳,而在应用神经营养因子的各组中又以ＢＤＮＦ＋ ＮＧＦ组为优,提示ＢＤＮＦ、ＮＧＦ能促进移植区中ＮＯＳ阳性神经元的发育生长,ＢＤＮＦ与ＮＧＦ联合使用可发挥协同作用。本文对ＢＤＮＦ和ＮＧＦ促进移植区中ＮＯＳ阳性神经元发育生长的机制进行了探讨。     

非胰岛素依赖型糖尿病人血浆内皮素水平与游离脂肪酸浓度的关系

为探讨非胰岛素依赖型糖尿病（Ｎｏｎｉｎｓｕｌｉｎ－ｄｅｐｅｎｄｅｎｔｄｉａｂｅｔｅｓｍｅｌｌｉｔｕｓ,ＮＩＤＤＭ）人内皮细胞损伤的可能机制,作者采用放射免疫法和反相高效液相色谱技术（ＨＰＬＣ）同时测定了３４例正常人和５６例ＮＩＤＤＭ病人血浆内皮素（ＥＴ）和游离脂肪酸（ＦＦＡ）浓度。结果显示：ＮＩＤＤＭ病人血浆ＥＴ和ＦＦＡ浓度较正常对照明显升高（均ｐ＜０．０１）,且血浆ＥＴ与空腹血糖（ＦＢＧ）水平和ＦＦＡ总浓度呈明显正相关（ｒ＝０．３２４和,ｒ＝０．３５１,均Ｐ＜０．０１）。结果提示：ＮＩＤＤＭ患者高血糖、脂肪酸代谢紊乱及胰岛素抵抗在其血管内皮损伤中可能有重要作用。     

大鼠室旁核和视上核NMDAR_1及NADPH-d阳性神经元分布的性别差异

为了研究ＮＭＤＡＲ１ 及ＮＡＤＰＨ－ｄ 阳性神经元在室旁核及视上核分布的性别差异,本研究采用雌雄Ｗ ｉｓｔａｒ 大鼠,恒冷箱切片,分别进行ＮＭＤＡＲ１ 免疫细胞化学和ＮＡＤＰＨ－ｄ 组织化学反应,并进行ＮＭ ＤＡＲ１ 免疫细胞化学和ＮＡＤＰＨ－ｄ 组织化学双重反应,光镜下观察室旁核及视上核的阳性细胞分布状况。观察结果发现,雄性组室旁核中的ＮＭＤＡＲ１ 和ＮＡＤＰＨ－ｄ 双重反应阳性细胞明显多于雌性组。ＮＭ ＤＡＲ１ 免疫反应组可见免疫阳性神经元,但雌雄两组之间未见明显差异。ＮＡＤＰＨ－ｄ 组化反应组可见高尔基样的阳性细胞,雌雄两组之间也未见明显差异。ＮＭ ＤＡＲ１、ＮＡＤＰＨ－ｄ 及ＮＭＤＡＲ１ 和ＮＡＤＰＨ－ｄ 双重反应各组在视上核未见性别差异。室旁核ＮＭＤＡＲ１ 和ＮＡＤＰＨ－ｄ 双重反应的分布有明显性别差异的研究结果表明,雄性大鼠室旁核可能通过ＮＭ ＤＡＲ－ＮＯ 途径介导雄性大鼠的某些性行为。     

大鼠第三脑室视前区NADPH-d阳性和NMDAR_1免疫阳性脑脊液接触神经元的形态(英文)

为探讨第三脑室接触脑脊液神经元在脑脊液与下丘脑之间信息传递过程中的作用以及脑脊液中谷氨酸对其影响的机制，本研究应用ＮＡＤＰＨ－ ｄ 组织化学方法、免疫组化方法以及组化双重反应方法研究了一氧化氮合酶和ＮＭ ＤＡＲ１ 在大鼠第三脑室视前区接触脑脊液神经元的表达。结果表明：一氧化氮合酶和ＮＭ ＤＡＲ１ 均在第三脑室接触脑脊液神经元表达，并且发现共存现象，在雌、雄大鼠间不存在明显的性别差异。尽管接触脑脊液神经元的生理功能尚不十分清楚，但本研究为接触脑脊液神经元作为脑－脑脊液环路的一部分、介导下丘脑对脑脊液中化学变化的感受提供了形态学依据，并提示谷氨酸－ＮＭ ＤＡＲ－Ｃａ２＋ －ＮＯ通路可能参与了这一过程。     

精神症状和单胺递质相互关系

应用高效液相色谱液对精神分裂症患者脑脊液中的ＤＡ、ＮＥ、Ａ、５－ＨＴ及其代谢产物进行测试，并和症状量表评分进行相关分析，发现ＢＰＲＳ评分和ＭＨＰＧ显著正相关，ＳＡＰＳ和ＭＨＰＧ、ＤＡ、ＨＶＡ显著正相关，ＳＡＮＳ与ＭＡＰＧ、ＤＡ、ＨＶＡ显著负相关，５－ＨＴ显著正相关。提示阳性症状可能与ＤＡ、ＮＥ功能亢盛有关，阴性症状可能与ＤＡ、ＮＥ功能低下及５－ＨＴ功能亢盛有关，但症状的变化和递质浓度的变化相关无显     

反义硫代寡核苷酸体外抑制柯萨奇病毒B3增殖的研究

目的 在ＣＶＢ３易感的Ｖｅｒｏ细胞系中观察与ＣＶＢ３ＲＮＡ５’ＮＣＲ的ｎｔ５８１－６０１区域区补的２１聚硫代ＡＯＤＮ抗病毒活性。方法 采用ＭＴＴ法测定细胞活性,直接观察ＣＰＥ,测定培养上清ＴＣＩＤ５０,并分别用ＥＬＩＳＡ和往点杂交法检测ＣＶＢ３抗原及其ＲＮＡ。结果 １．ＡＯＤＮ可推迟和减轻ＣＰＥ,且随ＡＯＤＮ浓度增加,对ＣＰＥ的抑制作用增强,感染细胞存活率也随ＡＯＤＮ浓度升高而升高;２．ＡＯＤＮ可     

大鼠隔外侧核和隔内侧核接受蓝斑一氧化氮合酶阳性神经元的投射

目的：探讨大鼠蓝斑核一氧化氮合酶(NOS)阳性神经元向隔外侧核(S1)和隔内侧核(Sm)的投射。方法：采用辣根过氧化物酶(HRP)逆行追踪法与还原型辅酶Ⅱ黄递酶(NADPH-d)组织化学法相结合的技术。结果：S1接受来自双侧蓝斑核NOS阳性神经元的投射,占蓝斑核向S1投射神经元的60％左右,同侧居多;双侧蓝斑核都有NOS阳性神经元投射至Sm,占蓝斑核向Sm投射神经元的55％左右。结论：S1和Sm接受蓝斑核NOS阳性神经元的投射,蓝斑核NOS阳性神经元至S1的投射呈同侧优势,至Sm的投射双侧无明显差别。     

脑创伤与缺血后大鼠脑内NOS阳性细胞变化和神经元损害的形态学观察

脑创伤与缺血后大鼠脑内ＮＯＳ阳性细胞变化和神经元损害的形态学观察李红丽蔡文琴（第三军医大学组胚教研室）本实验应用ＮＡＤＰＨ－ｄ组织化学技术结合尼氏染色观察了大鼠皮层针刺损伤及全脑缺血／再灌注术后皮层、海马及下丘脑室旁核等敏感区域神经元受损与ＮＯＳ阳性...     

Number of neurons in nucleus locus ceruleus in demented and non-demented patients: rapid estimation and correlated parameters

The total number of neurons in nucleus locus ceruleus (nLC) was counted in 20 patients with dementia (19 with dementia of the Alzheimer type) and 10 non-demented patients with the assistance of an image analyzer. The average number of neurons in a single representative section of nLC through the site of maximal neuronal density was then determined in each case. Both the mean total neuronal count and the average number of neurons in single sections correlated significantly with age, severity of dementia and length of nLC. The mean total number of neurons for demented (5571.700 +/- 4151.073) and non-demented (12533.700 +/- 2336.320) subjects differed significantly. The mean number of neurons derived by averaging counts of single representative sections from demented subjects (67.9 +/- 31.19) correlated significantly with the total number of nLC neurons determined by Quantimet-assisted analysis and differed significantly from the mean number of neurons derived by averaging counts of single representative sections of non-demented subjects of comparable age (110.0 +/- 16.59). With both methods of estimating neuronal numbers, the number of surviving nLC neurons in demented patients fell into 2 distinct groups, with cut-off points at 5,000 neurons (mean total counts) and 67 neurons (average of single representative section counts). These findings provide clear evidence that samples of the nLC neuronal population derived from single section counts produce data of comparable utility to those obtained from more laborious total neuronal counts in the human locus ceruleus.     

Role of noradrenergic terminals of the rat brain in passive-avoidance conditioning

Changes in the noradrenalin concentration in noradrenergic terminals were studied by a fluorescence-histochemical method in pars ventralis nuclei interstitialis striae terminalis, nucleus septi lateralis, and nucleus preopticus medialis in rats during passive-avoidance conditioning. In preliminarily trained animals the simple reproduction of the conditioned-reflex response (without electrical stimulation of the skin) led to a decrease in the noradrenalin concentration in all structures studied. The results indicate a role in the noradrenergic system of the brain in the reproduction of the passive-avoidance conditioned reflex.     

Increase in cytoplasmic free Ca2+ elicited by noradrenalin and serotonin in cultured local interneurons of mouse olfactory bulb.

Effects of noradrenalin and serotonin on cytoplasmic free Ca2+ concentrations ([Ca2+]i) were studied by using the fluorescent indicator fura-2 in cultured local interneurons of mouse olfactory bulb. Application of noradrenalin (0.1-100 microM) caused a rapid and concentration-dependent rise in [Ca2+]i, while isoproterenol was ineffective at concentrations up to 100 microM. The noradrenalin (1 microM)-induced increase in [Ca2+]i was completely inhibited by pretreatment with alpha 1-antagonist, prazosin (100 nM), whereas the inhibitory effect of alpha 2-antagonist, yohimbine, was about 100-times less potent. Serotonin (0.1-100 microM) also caused the dose-dependent rise in [Ca2+]i, which was inhibited by serotonin2 antagonist, ketanserin. Even in the absence of the extracellular calcium, the noradrenalin- or serotonin-induced increase in [Ca2+]i was observed. These results indicate that both noradrenalin and serotonin elicit the rise in [Ca2+]i in local interneurons of the olfactory bulb. They also suggest that the rise in [Ca2+]i is mediated by alpha 1-adrenergic and serotonin2 receptors, and that the increased calcium is mainly derived from intracellular calcium storage sites. The above results provide evidence to suggest that in the olfactory bulb, noradrenergic and serotonergic centrifugal fibers exert modulatory influences on synaptic interactions between mitral/tufted cells and local interneurons by increasing cytoplasmic Ca2+ in local interneurons.     

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.     

Activation of A1 and A2 noradrenergic neurons in response to running in the rat

Since running accompanied with blood lactate accumulation stimulates the release of adrenocorticotropic hormone (ACTH), running above the lactate threshold (LT) acts as stress (running stress). To examine whether A1/A2 noradrenergic neurons that project to the hypothalamus activate under running stress, c-Fos immunohistochemistry was used to compare the effects of running with or without stress response on A1/A2 noradrenergic neurons. Blood lactate and plasma ACTH concentrations significantly increased in the running stress group, but not in the running without stress response and control groups, confirming different physiological impacts between different intensity of running with or without stress. Running stress markedly increased c-Fos accumulation in the A1/A2 noradrenergic neurons. Running without stress response also induced a significant increase in c-Fos expression in the A1/A2 noradrenergic neurons, and the percentage of the increase was smaller than that of running stress. The extent of c-Fos expression in the A1/A2 noradrenergic neurons correlates with exercise intensity, signifying that this neuronal activation is running speed-dependent. We thus suggest that A1/A2 noradrenergic neurons are activated in response to not only running stress, but also to other physiological running, enhanced by non-stressful running. These findings will be helpful in studies of specific neurocircuits and in identifying their functions in response to running at different intensities.     

Antinociception induced by microinjection of substance P into the A7 catecholamine cell group in the rat.

Stimulation of neurons in the ventromedial medulla produces antinociception that is mediated in part by indirect activation of pontospinal noradrenergic neurons. Substance P-containing neurons located in the ventromedial medulla project to the A7 catecholamine cell group and may serve as an excitatory link between these two cell groups. Thus, the antinociception induced by stimulation of the neurons in ventromedial medulla may be mediated by substance P released from these projections which activates spinally projecting noradrenergic neurons in the A7 cell group. This hypothesis was tested by determining whether microinjection of various doses of substance P into the A7 cell group of the rat could induce antinociception. The results indicated that substance P induced dose-dependent antinociception that was more pronounced in the hindlimb ipsilateral to the microinjections. This observation is consistent with anatomical observations that noradrenergic A7 neurons project predominantly to the ipsilateral spinal cord dorsal horn. Moreover, the antinociceptive effects of substance P microinjection appear to be mediated at least in part by activation of spinally projecting noradrenergic neurons in the A7 cell group, because intrathecal injections of the alpha-2 noradrenergic antagonists yohimbine and idazoxan blocked these antinociceptive effects. The results of these experiments support the hypothesis that the antinociception induced by stimulation of neurons in the ventromedial medulla is mediated in part by activation of substance P-containing neurons that project to, and activate, spinally projecting noradrenergic neurons located in the A7 catecholamine cell group.     

Catecholaminergic innervation of guinea pig superior olivary complex

In mammals, olivocochlear neurons in the superior olivary complex project to the cochlea, providing input to outer hair cells and auditory afferents contacting inner hair cells. In the rat it has been demonstrated that olivocochlear neurons receive noradrenergic input, arising from the locus coeruleus and it has been demonstrated in this species using in vitro brain slices that noradrenaline exerts a direct, mostly excitatory effect on an olivocochlear subpopulation. The guinea pig is a more commonly used animal in auditory physiology than the rat and anatomical data on noradrenaline in the auditory brainstem in this species are lacking. Because it has been shown that a compact locus coeruleus is not present in the guinea pig, subtle species differences might be expected. Therefore, using immunohistochemical and tracing techniques we have investigated in the guinea pig (1) the noradrenergic and dopaminergic innervation of the superior olivary complex, (2) the anatomical relationship between noradrenergic fibres and olivocochlear neurons and (3) the origin of the noradrenergic input to this brainstem region. The results show that the guinea pig superior olivary complex receives moderately dense noradrenergic innervation and no dopaminergic innervation. In addition, noradrenergic fibres and varicosities were observed in close contact with both somata and dendrites of olivocochlear neurons, strongly suggestive of synaptic contacts. Finally the results show that a significant component of the noradrenergic innervation of the guinea pig superior olivary complex arises in the locus subcoeruleus, which is a structure likely to be the homologue of the locus coeruleus in rats and other species.     

Microinjected morphine suppresses the activity of locus coeruleus noradrenergic neurons in freely moving cats

Microinjection of morphine (1.0 microgram/0.1 microliter) produced a significant suppression (approximately 60%) of the single unit activity of locus coeruleus noradrenergic neurons in freely moving cats. This effect was reversible by systemic administration of the opioid receptor antagonist, naloxone (1.0 mg/kg i.v.). The microinjection of naloxone (1.0 microgram/0.1 microliter), however, was without effect on the spontaneous activity of noradrenergic neurons in the locus coeruleus. Non-noradrenergic neurons recorded in the same vicinity showed no consistent response to the microinjection of morphine. These results suggest that the direct effect of opioids in the locus coeruleus is an inhibition of noradrenergic neuronal activity. Furthermore, it appears that opioid influences upon these neurons are not tonically active.     

Neuronal composition of the intramural ganglia of the gastro-intestinal tract

The neuronal composition of the intramural plexuses of the gastro-intestinal tract in cats was studied by luminescence microscopy. The intramural ganglia were found to contain three populations of neurons. The neurons of the first population showed no specific catecholamine fluorescence. Those of the second population had fluorescence of the perikaryon and processes. The perikarya of the third neuron population were not fluorescent but adrenergic synapses were seen on their surface. Intraperitoneal injection of exogenous noradrenalin (1 mg) led to an increase in the number of neurons of the third population with specific fluorescence. On this basis it was concluded that as well as parasympathetic intramural neurons the intramural ganglia of the gastro-intestinal tract also contain sympathetic cells, most of which exhibit specific fluorescence only after their mediator content has been artificially increased.     

Noradrenergic and non-noradrenergic nerves containing small granular vesicles in Auerbach's plexus of the guinea-pig: evidence against the presence of noradrenergic synapses

The appearance and distribution of varicosities containing small granular vesicles in Auerbach's plexus of the guinea-pig ileum, distal colon and rectum has been studied with the electron-microscope. Two types of varicosity were recognised. The first type was located predominantly at the surface of the plexus and did not form synapses on intrinsic neurons. This type became labelled with 5-hydroxydopamine, a specific marker for noradrenergic axons, and was destroyed by 6-hydroxydopamine and extrinsic denervation, procedures which lead to degeneration of noradrenergic nerves in the gut. The second type formed axodendritic and axosomatic synapses on intrinsic neurons and the morphology of its synaptic vesicles differed subtly from that of the first type. The second type was unaffected by 5-hydroxydopamine, 6-hydroxydopamine, or extrinsic denervation. It is concluded that the two types of small granular vesicle-containing varicosities belong to different neurons and that the first type is noradrenergic. Noradrenergic varicosities do not, therefore, form synapses in Auerbach's plexus. This conclusion is in accord with the electrophysiological findings. The second type of small granular vesicle-containing varicosity is not noradrenergic although it was formerly thought to be so. It is intrinsic to the gut and is resistant to the serotoninergic neurotoxin, 5,6-dihydroxytryptamine.