大鼠脑干面口部运动神经元与5—HT样,SP样,L—ENK样,GABA样和 …

为了观察５－羟色胺样、Ｐ物质样、亮氨酸－脑啡肽样，γ－氨基丁酸和甘氨酸样阳性终末与脑干面口部运动核间的突触联系，用ＨＲＰ逆行追踪免疫组织化学染色相结合的双标技术，在电镜下对５－ＨＴ样、ＳＰ样、Ｌ－ＥＮＫ样、ＧＡＢＡ样和Ｇｌｙ样阳性终末与ＨＲＰ逆标面口部运动神经元间的突触联系进行了观察。５－ＨＴ样、ＳＰ样、Ｌ－ＥＮＫ样和Ｇｌｙ样阳性终末在脑干面口部运动内呈均匀分布，没有区域特异性。将ＨＲＰ注入面口部     

大鼠脑干5-羟色胺能和儿茶酚胺能神经元向孤束核的投射——荧光金标记结合免疫荧光技术研究

应用荧光金逆行追踪和免疫荧光技术相结合的方法,对投射至大鼠孤束核的５－羟色胺能和儿茶酚胺能神经纤维的脑干来源进行了研究。结果证明,将荧光金注入一侧孤束核的中尾段,逆标神经元分布于脑干下行抑制系统的大部分核团。结合５－羟色胺和酪氨酸羟化酶免疫荧光组织化学技术研究发现：荧光金／５－ＨＴ 双重反应阳性神经元分布于中脑导水管周围灰质的腹外侧区、中缝背核、脑桥被盖网状核、脑桥尾侧网状核、中缝桥核、中缝正中核、中缝大核、巨细胞网状核α部、中缝隐核和中缝苍白核等核团,其中中央灰质腹外侧区、中缝大核、巨细胞网状核α部、中缝隐核、中缝苍白核等处的荧光金／５－ＨＴ 双重反应阳性神经元数量占脑干向孤束核投射的５－ＨＴ 阳性神经元总数的近７０％ （６９．９１％ ）;荧光金／酪氨酸羟化酶双重反应阳性神经元主要分布于脑干中脑中央灰质腹外侧区、Ａ７、蓝斑及蓝斑下核（Ａ６）、Ａ５、巨细胞网状核α部和Ａ１ 等核团,其中中央灰质腹外侧区、Ａ５、巨细胞网状核α部和Ａ１ 内的荧光金／酪氨酸羟化酶神经元数量占脑干向孤束核投射的酪氨酸羟化酶阳性神经元总数的８８．１７％ 。本研究提示,大鼠脑干下行抑制系统形成了至ＮＴＳ中尾段的以中脑中央灰质腹外侧区及中缝大核、?     

大鼠延髓内脏带的化学神经解剖学

应用免疫组织化学技术（ＰＡＰ法和ＡＢＣ法），对儿茶酚胺类（以酪氨酸羟化酶０ＴＨ作标记物）、５－羟色胺、胆碱类（以胆碱乙酰化酶－ＣｈＡＴ作标记的）以及神经肽类（Ｐ物质、降钙素基因相关肽、神经降压素、胆膈 、生长抑素和神经肽Ｙ）等物质在ＳＤ大鼠延髓内脏带内的分布进行了观察。结果证明，上述物质均较集中地分布于延髓内脏带，即位于延宫尾段（从闩尾侧１．０ｍｍ到闩则０．８ｍｍ）的一条从人则至腹外     

大鼠中缝背核三种递质传入纤维的脑干起源—免疫细胞化学和酶组织…

实验用ＨＲＰ逆行追踪法和免疫细胞化学ＡＢＣ法，研究了大鼠中缝背核内５－羟色胺，Ｐ物质以及亮脑啡肽三种传入纤维的脑干起源。发现中央上核，中缝大中脑导水管周围灰质，脚间核和第Ⅴ脑室中央灰质均有数量不等的５－羟色胺，Ｐ物质和亮脑啡肽能神经元向中缝背核投射。     

大鼠孤束核内降钙素基因相关肽能神经的分布及其与儿茶酚胺能神经元的关系

用双重免疫染色包埋前免疫电镜法，在光镜和电镜水平观察了大鼠孤束核内降钙素基因相关肽神经元的分布及其神经末梢与儿茶酚胺能神经元之间的关系。降钙素基因相关肽阳性神经元呈棕色，主要分布于孤束核的连合亚核、内侧亚核以及背侧周边区。在孤束核的尾侧部、连合亚核的腹外侧部及背侧周边区观察到蓝绿色的酪氨酸羟化酶阳性神经元和棕色的降钙素基因相关肽阳性神经元和神经纤维，其中一些神经元内两者共存。电镜观察结果：酪氨酸羟化酶阳性神经元细胞质内有棒状或卵圆形的电子密度较高的吲哚-β-半乳糖苷酶反应产物，降钙素基因相关肽阳性神经末梢内见到电子密度较高的DAB反应产物以及清亮小泡和颗粒小泡。降钙素基因相关肽阳性神经末梢和酪氨酸羟化酶阳性神经元之间形成对称性轴一体突触。     

大鼠延髓内含前—原脑啡肽mRNA,甲硫氨酸—脑啡肽和亮氨酸—脑啡肽神…

应用原位杂交和免疫组织化学方法，对成年大鼠延髓内含前－原脑啡肽（ＰＰＥ）ｍＲＮＡ和甲硫氨酸－脑啡肽（Ｍ－ＥＮＫ）与亮氨酸－脑啡肽（Ｌ－ＥＮＫ）免疫反应神经元进行观察。结果表明：含ＰＰＥｍＲＮＡ的神经元胞体，多数分布在孤束核、腹外侧压以及两者之间的网状结构等形成的一条从背内侧到腹外侧区的弧形带内。Ｍ－ＥＮＫ与Ｌ－ＥＮＫ样免疫反应阳性结构（神经元胞体、纤维和终末）也主要密集分布在该带内。本结果对ＥＮＫ     

大鼠孤束核内降钙素基因相关肽能神经的分布及其与儿茶酚胺 …

用双重免疫染色包埋前免疫电镜法，在光镜和电镜水平观察了大鼠孤束核内降钙素基因相关肽神经元的分布及其神经末梢与儿茶酚胺能神经元之间的关系。降钙素基因相关肽阳性神经元呈棕色，主要分布于孤束的连合亚核、内侧亚核以及背侧周边区。在孤束核的尾侧部、连合亚核的腹外侧部及背侧周边区观察到蓝绿色的酪氨酸羟化酶阳性神经元和棕色的降钙素基因相关肽阳性神经元和神经纤维，其中一些神经元内两者共存。电镜观察结果：酪氨酸羟化     

一氧化氮合酶在自发性高血压大鼠视上核及室旁核的分布

目的：探讨自发性高血压大鼠视上核及室旁核内一氧化氮合酶阳性神经元的改变。方法：用ＮＡＤＰＨ－ｄｉ－ａｐｈｏｒａｅ组织化学方法,观察和比较了一氧化氮合酶在自发性高血压大鼠和正常对照大鼠视上核和室旁核的分布间下观察下捕脑视上核及室旁核的一氧化氮合酶阳性细胞形态、分布并进行计数。用图像分析仪测其灰度显差异。视上核及室旁核内的一氧化氮合酶阳性细胞灰度值在高血压组均明显高于对照组。提示这两上核内一氧化氮合酶     

癫痫发作的大鼠延髓内脏带神经元的FOS表达

为研究大鼠"延髓内脏带"神经元在红藻氨酸诱导癫痫发作时的反应,本文用抗FOS蛋白和抗酪氨酸羟化酶双重免疫组织化学方法,对下列三组实验动物进行了观察：（1）红藻氨酸（10mg／kg）腹膜腔注射组;（2）L－精氨酸慢性腹膜腔注射给药（40mg／kg,2次／d,共4d）后,再注射红藻氨酸组;（3）L－硝基精氨酸慢性腹膜腔注射给药（50mg／kg,2次／1d,4d）后,再注射红藻氨酸组。所有动物均在红藻氨酸注射3h后处死,制片,染色。结果：红藻氨酸注射组动物出现癫痫发作,"延髓内脏带"内显示密集的FOS阳性神经元,其中FOS／酪氨酸羟化酶神经元占酪氨酸羟化酶神经元总数的85％,占FOS阳性神经元总数的15％;酪氨酸羟化酶神经元出现胞体变形,胞浆内有空泡,突起短缺等现象。L－精氨酸＋酪氨酸羟化酶组癫痫发作较轻,"延髓内脏带"内FOS阳性神经元较少;L-硝基精氨酸＋酪氨酸羟化酶组癫痫发作加重,死亡率增高,"延髓内脏带"内FOS阳性神经元极多。以上结果表明,"延髓内脏带"内神经元在癫痫发作时反应强烈;一氧化氮介质可能有抗惊厥作用。     

一氧化氮合酶在自发性高血压大鼠脊髓中间带灰质的分布

目的 探讨自发性高血压大鼠脊髓中间带灰质内一氧化氮合酶阳性细胞和纤维的分布及其与自发性高血压的关系。方法 采用ＮＡＤＰＨ－ｄ组织化学方法,并以计算机图像分析系统对脊髓胸腰部中间外侧核本部区ＮＡＤＰＨ－ｄ染色强度（Ａ值）进行半定量分析。结果１．在脊髓中间带灰质区,一氧化氮合酶阳性神经细胞和纤维主要见于胸腰部中间外侧核本部区和中央管周围,在中介核区也有少量分布。胸腰部中间外侧核本部区染色阳性细胞成群分布,细胞群的大小、间距随节段而稍有不同。一氧化氮合酶阳性细胞分布模式在自发性高血压大鼠和正常鼠两组之间未发现明显差异。２．在脊髓Ｔ２、Ｔ５、Ｔ８、Ｔ１１、Ｌ１各节段中间外侧核本部区,一氧化氮合酶阳性物质的Ａ值,自发性高血压大鼠比正常大鼠低。统计结果显示,差异有显著性（Ｐ〈０．０５）。结论 脊髓中间带灰质的交感神经节前     

Monoamine contents and norepinephrine turnover in brain stem nuclei of young and adult spontaneously hypertensive and Wistar-Kyoto rats

To investigate the brain stem monoamine mechanism in the development and maintenance of hypertension of spontaneously hypertensive rats (SHR), we determined monoamine contents and norepinephrine turnover in discrete brain stem nuclei which are known to relate with cardiovascular control. Specific areas and brain stem nuclei were dissected from serial frozen slices of 300 microns thickness according to the atlas of Palkovits and Jacobowitz. The dissected tissues were homogenized, centrifuged and the supernatants were injected into high performance liquid chromatography with electrochemical detection (HPLC-ECD). Norepinephrine (NE), dopamine (DA), serotonin (5-HT), and 5-hydroxyindoleacetic acid (5-HIAA) contents were determined. NE turnover was also determined 2 hour after alpha-methyl-p-tyrosine administration (250 mg/kg, i.p.). In 4-week old SHR, the only significant change observed was decreased NE contents in the nucleus tractus solitarii (NTS). Such decreases in NE contents of the NTS were also found in 8- and 16-week old SHR. However, there were no differences in NE turnover in the NTS between SHR and WKY. Increased NE contents were found in the A1, A5, and nucleus reticularis gigantocellularis (RG) in the later stages (8 and 16 weeks) in SHR. Furthermore, increased NE turnover was seen in the RG of SHR at 16-week old, indicating increased neuronal activity. Dopamine, 5-HT and 5-HIAA showed no consistent changes between SHR and WKY. Increased NE levels were observed in later stages after development of hypertension, suggesting the increased NE in adult SHR may represent a central adaptive change secondary to the established hypertension. Since increased NE levels were consistently found in or around the regions which are known as vasomotor centers, we assume that these increased NE might serve to maintain hypertension or to inhibit a further increase in blood pressure. In contrast, the NE contents were decreased with constant turnover in NTS of SHR aged 4, 8, and 16 weeks. Constant turnover in NE could not compensate for reduced NE in NTS and may lead to a functional reduction or reduced noradrenergic activity. This defect in intrinsic noradrenergic neurons in NTS may trigger the development of genetic hypertension in SHR. In conclusion, the present results demonstrate that NE levels of SHR in the NTS were consistently decreased compared with those of WKY in all age groups. In later stages, increased NE levels were observed in A1, A5 and RG of SHR. These results indicate that brain stem monoamine system, especially noradrenergic neurons, contributes to the development and maintenance of hypertension in SHR.     

P物质在自发性高血压大鼠脊髓中间带灰质的分布

为探讨自发性高血压的发病机理，本实验用免疫细胞化学方法结合体视学分析，研究了自发性高血压大鼠脊髓中间带灰质内Ｐ物质免疫反应阳性纤维和终末的分布。结果如下：（１）Ｐ物质免疫反应产物在自发性高血压大鼠和京都种正常血压大鼠脊髓中间带灰质分布的模式未见明显差异；（２）在脊髓Ｔ２、Ｔ５、Ｔ８、Ｔ１１、Ｌ１ 各节段的中间外侧核本部区的平均光密度值，自发性高血压大鼠高于京都种正常血压大鼠（Ｐ＜ ０．０５）。这一结果提示，在自发性高血压大鼠，脊髓胸腰部中间外侧核本部区Ｐ物质免疫反应阳性产物表达增加。而该区Ｐ物质表达的增加意味着该区交感节前神经元机能状态的某种改变，这种改变可能与自发性高血压大鼠血压的升高和维持有关。     

Brainstem oxytocinergic modulation of heart rate control in rats: effects of hypertension and exercise training

Oxytocinergic brainstem projections participate in the autonomic control of the circulation. We investigated the effects of hypertension and training on cardiovascular parameters after oxytocin (OT) receptor blockade within the nucleus tractus solitarii (NTS) and NTS OT and OT receptor expression. Male spontaneously hypertensive rats (SHR) and Wistar–Kyoto (WKY) rats were trained (55% of maximal exercise capacity) or kept sedentary for 3 months and chronically instrumented (NTS and arterial cannulae). Mean arterial blood pressure (MAP) and heart rate (HR) were measured at rest and during an acute bout of exercise after NTS pretreatment with vehicle or OT antagonist (20 pmol of OT antagonist (200 nl of vehicle)^–1). Oxytocin and OT receptor were quantified (³⁵S-oligonucleotide probes, in situ hybridization) in other groups of rats. The SHR exhibited high MAP and HR ( P < 0.05). Exercise training improved treadmill performance and reduced basal HR (on average −11%) in both groups, but did not change basal MAP. Blockade of NTS OT receptor increased exercise tachycardia only in trained groups, with a larger effect on trained WKY rats (+31 ± 9 versus +12 ± 3 beats min⁻¹ in the trained SHR). Hypertension specifically reduced NTS OT receptor mRNA density (–46% versus sedentary WKY rats, P < 0.05); training did not change OT receptor density, but significantly increased OT mRNA expression (+2.5-fold in trained WKY rats and +15% in trained SHR). Concurrent hypertension- and training-induced plastic (peptide/receptor changes) and functional adjustments (HR changes) of oxytocinergic control support both the elevated basal HR in the SHR group and the slowing of the heart rate (rest and exercise) observed in trained WKY rats and SHR.     

脊髓侧角区5-HT、TH、SP和L-ENK免疫反应纤维和终末的超微结构

本文应用免疫细胞化学ABC法,在电镜下观察脊髓侧角区单胺能和某些肽能纤维及末梢的突触组合。大鼠侧角内的5-HT、TH、SP和L-ENK免疫反应纤维均为无髓纤维。在侧角细胞簇内,这些纤维穿行于胞体之间,有的与胞体相邻,但很少与胞体形成轴-体突触。这些单胺和肽类纤维也与树突伴行,在树突束内数量最多。有时一小束无髓纤维都含同一种免疫反应物质。轴-树突触是各种免疫反应纤维终末所形成突触的主要形式。各种纤维终末所含的小泡多为圆清亮小泡,或兼有少数大颗粒泡。SP和L-ENK纤维膨体内的小泡与其终末内者不同,大颗粒泡较多,有时约占半数。各种免疫反应终末所组成参与的突触,对称或非对称型均不显现优势。     

Pathophysiologic role of natriuretic peptides

To evaluate the pathophysiologic role of atrial natriuretic peptide (ANP) in hypertension, hemodynamic effects of human ANP and antiserum against rat ANP were investigated in spontaneously hypertensive rats (SHR) and Wistar-Kyoto rats (WKY). Intravenous administration of human ANP caused greater hypotension associated with a decrease of cardiac output in SHR than in WKY, which suggests that SHR have enhanced responsiveness to exogenous ANP. The antiserum increased blood pressure and cardiac output, with the latter being significantly greater in SHR than in WKY. These results suggest that endogenous ANP counteract, in part, the maintenance of hypertension. In addition, hemodynamic and renal excretory effects of brain natriuretic peptide (BNP), a novel natriuretic peptide identified from porcine, were studied in SHR and WKY. BNP caused marked natriuresis and hypotension in a dose-dependent fashion, as observed with ANP. Not only ANP but also BNP may have a role in the regulation of blood pressure and water-electrolyte balance.     

猫延髓吻侧腹外侧区5-羟色胺、苯乙醇胺氮位甲基转移酶、P物质和亮氨酸-脑啡肽免疫反应神经元的分布

用免疫组织化学ABC技术,观察了猫延髓吻侧腹外侧区(RVL)5-羟色胺(5-HT)、苯乙醇胺氮位甲基转移酶(PNMT)、P物质(SP)、亮氨酸-脑啡肽(L-ENK)免疫反应神经元的分布,并用免疫荧光组织化学法结合荧光素逆行追踪法初步研究了RVL内5-HT、SP、L-ENK免疫反应神经元向胸髓的投射。结果表明:4种物质的免疫反应神经元主要分布在旁巨细胞外侧核的尾侧部和外侧网状核的吻侧部。SP阳性细胞还见于疑核腹外侧紧邻的网状结构中。有的5-HT、SP和L-ENK阳性细胞位于紧靠软脑膜的区域。在RVL的大部分区域,4种物质的免疫反应细胞呈重叠分布。在斜方体尾侧1.0～3.5mm平面,距中线约3.3mm,距腹面约0.8mm的区域,5-HT、PNMT和L-ENK阳性神经元均聚集成团,突起相互交织,在吻尾方向上各自形成长短不一的连续细胞柱。这3条细胞柱在斜方体尾侧1.5～2.5mm平面几乎完全重合。RVL部分5-HT、SP和L-ENK神经元发出纤维投射至胸髓。本文还就上述物质在此区的功能意义进行了讨论。     

自发性高血压鼠脑底动脉单胺能神经分布的形态学观察

应用免疫组织化学ABC法，对10只雄性自发性高血压大鼠和10只正常大鼠脑底动脉单胺能神经纤维分布进行了观察、在自发性高血压鼠脑底血管的大脑前动脉、大脑中动脉和基底动脉壁上均可见棕褐色的免疫反应阳性纤维，纤维较粗，弯曲，环行走行，密度较高。与正常鼠同一部位脑底动脉壁的阳性纤维密度比较有明显增加。经统计学检验，P＜0．05，表明高血压鼠和正常鼠脑底动脉之间的单胺能神经纤维的密度有明显差异。本文结果提示，高血压鼠脑底动脉各主要分支较正常鼠脑底动脉各主要分支有更高密度的单胺能神经纤维分布，这些纤维属于交感神经纤维，说明交感神经可能在高血压鼠的脑血管活动和对高血压的发生和发展具有重要的作用。     

不同年龄高血压大鼠血管平滑肌中ERK和MKP-1的表达

目的：研究不同年龄的自发性高血压大鼠（SHR）和Wistar Kyoto大鼠（WKY）主动脉平滑肌中丝裂原活化蛋白激酶（MAPK）及其磷酸酶（MKP-1）的表达及其与高血压的关系。 方法： 用tail-cuff测量大鼠尾动脉血压；分别用Western blotting法和RT-PCR法半定量测定血管平滑肌中磷酸化细胞外信号调节激酶（p-ERK）和MKP-1的蛋白表达以及MKP-1 mRNA的含量。 结果： （1）SHR的血压自8周龄起明显高于WKY（P<0.01），且随年龄增长而升高（P<0.05）至14周以后趋于稳定；（2）SHR主动脉平滑肌中的p-ERK表达明显高于同年龄的WKY（P<0.01），随年龄增长而递增（P<0.05），与血压呈正相关；（3）SHR主动脉平滑肌中MKP-1蛋白明显高于同龄WKY，而mRNA的表达在5周龄时明显高于WKY，之后均随年龄的增长而递减（P<0.05），与血压和ERK呈负相关，而WKY下降不明显。 结论： MKP-1在高血压的发生和发展过程中起重要作用，其表达逐渐下降可能是导致ERK激活增加，从而导致血管平滑肌细胞增殖、血压升高的重要原因。     

Alterations in tissue concentration and gene expression of adrenomedullin in spontaneously hypertensive rats

In order to elucidate the role of adrenomedullin in hypertension, we have compared concentrations of immunoreactive rat adrenomedullin and adrenomedullin messenger RNA levels in tissues of 8-week-old spontaneously hypertensive rats (SHR) with those of age-matched Wistar-Kyoto rats (WKY). The adrenomedullin immunoreactivity concentrations in adrenal gland and cardiac atrium were significantly higher in SHR than in WKY. The adrenomedullin content of cardiac ventricle was also significantly higher in SHR than in WKY. The rat adrenomedullin messenger RNA levels in adrenal gland and heart of SHR were also higher than those of WKY. These results suggest that adrenomedullin participates in the mechanism to counteract the blood pressure elevation in SHR.