中枢nesfatin-1对大鼠夜间摄食和胃排空的影响

目的:观察中枢nesfatin-1对大鼠夜间摄食和胃排空的影响。方法:大鼠经腹腔注射硫酸仲丁巴比妥(100~150 mg/kg)麻醉,侧脑室、第四脑室或小脑延髓池注射nesfatin-1或CRF受体拮抗剂astressin-B或astressin2-B,观察对摄食、胃排空的影响。结果:侧脑室注射nesfatin-1后大鼠第3-6 h夜间进食量(t=3.05~3.58,P0.01)和3 h和6 h的累积进食量(t=5.90~12.1,P0.01)明显减少,nesfatin-1的该抑制效应可被预先侧脑室注射astressin-B或astressin2-B阻断(t=1.06~2.22,P0.05)。第四脑室或小脑延髓池注射nesfatin-1后大鼠夜间摄食量在第1h就明显减少(t=2.59~6.26,P0.05~0.01),持续减少至5-6h(t=1.69~7.42,P0.05~0.01)。侧脑室注射不同剂量nesfatin-1(0.05或0.5μg)20 min后GE率明显降低,且随注射剂量增大,GE率越低(t=3.25~4.67,P0.01)。若预先给予大鼠CRF受体拮抗剂astressin2-B(30μg)再注射nesfatin-1(0.5μg),nesfatin-1抑制大鼠胃排空效应明显减弱(t=2.45~2.85,P0.05)。禁食24 h后再喂食2 h,大鼠下丘脑中nesfatin-1表达明显增加(t=2.87,P0.05),禁食24 h后血浆nesfatin-1水平明显降低(t=1.51,P0.05)。结论:Nesfatin-1抑制摄食作用可能由nesfatin-1和CRF2信号系统共同调节。     

侧脑室注射氟安定对家兔摄食的影响

口服或外周注射安定、乙磺氟安定、氟安定等苯二氮董化合物(BZs)可促进大鼠等动物摄食。BZs可加强中枢GABA能神经抑制效应,因此BZs促进动物摄食是否是同脑内GABA能有关的中枢作用,本实验通过观察侧脑室注射氟安定对家兔摄食的影响,就这方面作了初步探讨。     

侧脑室注射orexin-A对大鼠昼夜摄食的影响

目的:探讨侧脑室注射orexin-A对大鼠昼夜摄食的影响。方法:将Wistar大鼠随机分组,采用单剂量侧脑室注射和连续侧脑室注射以及外周注射法,分别于日间和夜间给药,测量大鼠24小时内各阶段的摄食量以及相应生化指标。结果:在光照期间,侧脑室微量注射orexin-A,大鼠4小时内摄食量显著增加(P0.05),且呈剂量依赖关系(P0.05)。在夜间初期(18:00)侧脑室注射orexin-A,大鼠食物摄入量无显著差异(P0.05)。但在中午12:00给予侧脑室注射orexin-A,注射后4小时内大鼠摄食量显著高于NS对照组(P0.05)。连续8日给予orexin-A侧脑室注射,可使注射后日间摄食量显著增加(P0.05),而夜间摄食量显著减少(P0.05),但24小时内总的摄食量不变(P0.05)。orexin-A并未改变棕色脂肪组织温度、末梢血糖、血浆瘦素等指标的水平。结论:orexin-A对大鼠摄食的调节具有昼夜节律性。     

大鼠侧脑室内注射组胺对颈动脉窦压力感受性反射的影响及其中枢机制

在50只麻醉的大鼠孤离双侧颈动脉窦区,将不同窦内压（ISP）与其对应的平均动脉压（MAP）值进行Logistic五参数曲线拟合,根据所得ISP-MAP关系曲线及其特征参数,观察侧脑室注射（i.c.v)组胺（HA）对颈动脉窦压力感受性反射（CSR）的影响,并对其作用机制进行了初步探讨。结果如下：（1）i.c.v.HA(100ng)导致ISP-MAP关系曲线显著上移,ISP和增益（Gain)关系曲线中部明显下移,反射参数中阈压（TP）,饱和压（SP）和最大增益时的窦内压（ISPGmax)值增大,MAP反射变动范围（MAPrange)及反射最大增益（Gmax)减小。（2）预先i.c.v.H1受体拮抗剂氯苯吡胺（CHL,5μg）或H2受体拮抗剂西咪替丁（CIM,15μg）,可明显减弱HA的上述效应,CIM的这种减弱作用不如CHL的显著。（3）预先同时i.c.v.CHL和CIM（分别为5和15μg）,能完全取消HA的效应。（4）预先向孤束核（NTS）内注射CHL（0.5μg）或CIM（1.5μg）,对HA效应的影响与i.c.v.CHL或CIM后的相类似,但NTS内注射CHL或CIM后i.c.v.HA所致的TP变化表现明显下降。以上结果提示,侧脑室给HA使CSR产生快速重调定,反射敏感性下降,功能受抑;其机制是通过中枢HA受体介导,H1受体的作用比H2受体更为明显;下丘脑-NTS的HA能通路可能是HA调节CSR的下行通路之一。NTS处的HA受体在i.c.v.HA抑制CSR的机制中可能发挥重要的作用。     

侧脑室注射微量促甲状腺素释放激素对大鼠胃运动的影响

促甲状腺素释放激素(TRH)是一种重要的脑肠肽,它除了有激素的作用外,还可作为神经递质或调质发挥多种生物学功能。目前对TRH调节胃肠运动功能的研究较少。本研究规察侧脑室内注射微量TRH对胃运动的影响,并初步探讨了其作用机理。     

中枢注射抗Orexin抗体对禁食大鼠摄食的抑制作用

目的:探讨中枢注射抗Orexin抗体对禁食大鼠摄食的抑制作用。方法:采用免疫组织化学法和蛋白质免疫印迹分析法,对Orexin抗体的特异性进行了检测,分析Orexin阳性神经元和阳性神经纤维在大脑中的分布。给予24 h禁食大鼠中枢注射抗Orexin抗体,计算其对大鼠食物摄入量的影响。结果:蛋白质免疫印迹分析显示,Orexin抗体能够检测到合成的Orexin-A。免疫组织化学分析显示,Orexin阳性神经元存在于外侧下丘脑区域和穹窿周核,Orexin阳性神经纤维大量投射至弓状核、下丘脑室周核和下丘脑室旁核、菱形丘脑核、丘脑室旁核、缰内侧核和纹质丘脑、中脑的中央灰质区、蓝斑核和中缝核、脑桥和髓质的网状结构、对疑核和迷走神经复合体。与注射羊血清相比,给予45μg/10μL抗Orexin抗体侧脑室注射则抑制了大鼠的食物摄入量(P0.05)。高剂量抗Orexin抗体能显著地抑制大鼠摄食,并且呈剂量依赖关系(P0.05)。结论:中枢注射抗Orexin抗体对禁食大鼠摄食具有抑制作用,并呈剂量依赖关系。     

伏隔核微注射orexin-A 对大鼠摄食和活动的影响

目的:探讨伏隔核微注射orexin-A后,大鼠摄食和活动的变化。方法:采用SD大鼠(250-280g),用脑立体定位仪在伏隔核植入微量注射管。大鼠随机分组,分别微注射乳酸格林液(Ringer’s),orexin-A 100pmol和500pmol。观察微注射后大鼠0-1h,1-2h,2-4h摄食和0-30min,30-60min,60-90min,90-120min活动性变化。结果:Orexin-A微注射后,大鼠0-1h,1-2h摄食量增加;30-60min,60-90min,90-120min的活动性显著增加(P<0.05 vs对照组)。结论:伏隔核是orexin-A刺激大鼠增加摄食量,提高其活动性的作用点。     

侧脑室内注射复方丹参注射液对大鼠肺动脉压的影响

复方丹参注射液 (ｓａｌｖｉａｌｍｉｌｔｉｏｎｒｒｈｉｊａｅ,ＳＭ) ,具有扩张血管 ,改善微循环 ,护肝抗凝等作用 ,临床上主要用于治疗心绞痛 ,心肌梗塞等疾病 ,关于ＳＭ降低肺动脉压的作用早有报道 ,但其对中枢作用尚未见报道 ,本研究主要是从侧脑室注射微量ＳＭ以观察其对肺动脉压的作用。1　材料与方法(1)材料　实验用药均用新鲜生理盐水配制 ,丹参注射液为十堰康迪制药厂生产 (ＳＭ ,ｐＨ 6 .8) ,心得安 (Ｐｒｏｐｒａｎｏｌｏｌ,ｐＨ6 .8) ,酚妥拉明 (Ｐｈｅｎｔｏｌａｍｉｎｅ ,ｐＨ ,6 .8) ,阿托品 (Ａｔｒｉｏｐｉｎｇ ,ｐＨ6 .8)…     

芹菜素对雌性大鼠生殖轴的影响

目的：探讨芹菜素在10-9mol/L浓度时对雌性大鼠生殖功能的影响。方法：应用侧脑室注射方法观察芹菜素对雌性大鼠生殖轴激素含量的影响。在侧脑室注射后第3天取血浆,采用放免技术测定血浆中促性腺激素释放激素（GnRH）、卵泡刺激素（FSH）、黄体生成素（LH）、雌二醇（E2）、孕酮（P）的含量。结果：在给药后第3天血浆中促性腺激素释放激素（GnRH）、卵泡刺激素（FSH）、黄体生成素（LH）的含量增加而雌二醇（E2）、孕酮（P）的含量降低,差异有显著性。结论：芹菜素抑制雌二醇（E2）合成中芳香化酶的活性实现对雌二醇（E2）、孕酮（P）分泌的抑制。芹菜素通过影响雌激素受体而使下丘脑的促性腺激素释放激素（GnRH）和腺垂体的卵泡刺激素（FSH）、黄体生成素（LH）分泌增加。     

一种新的摄食相关肽——obestatin

最近从大鼠胃组织中发现一种新的摄食相关肽,命名为obestatin。Obestatin是一种ghrelin相关肽,与ghrelin来自同一基因。Obestatin由23个氨基酸组成,分子量2516．3,氨基酸序列为FNAPFDVGIKLSGAQYQQHGRALNH2,C末端甘氨酸残基带有酰胺化修饰基团。Obestatin是G蛋白偶联孤儿受体（GPR39）的内源性配体。功能研究表明,obestatin与ghrelin截然相反,给小鼠腹腔或脑室注射obestatin,均呈时间和剂量依赖性抑制摄食：抑制大鼠的体重增加;持续件抑制冒排卒;减少宰肠肌条的收缩活动,     

大鼠侧脑室注射瘦素对卵巢摘除给予雌激素子宫腔液2-DE图谱分析

瘦素(leptin)通过动物下丘脑-垂体轴对其生殖活动予以调节,但瘦素通过下丘脑对子宫调节作用仍不清楚.本研究在大鼠侧脑室微量注射瘦素6 h后,采用双向凝胶电泳(2-DE)结合基质辅助激光解吸电离飞行时间质谱(MALDI-TOF-MS)技术分离、鉴定了卵巢摘除给予雌激素的子宫腔液蛋白质表达谱.我们获得了分辨率较高、重复性较好的2-DE图谱|质谱分析结合SwissProt蛋白数据库检索,共鉴定了24个差异表达的蛋白质与生理盐水注射（对照）比较,侧脑室微量注射瘦素引起22个蛋白质分子显著上调,2个显著下调.重要的是,在上调的蛋白质分子中包括转铁蛋白、基质金属蛋白酶7、膜联蛋白A1、补体B因子、磷酸甘油酸酯变位酶1、烯醇酶、冠蛋白-1A等15个蛋白质分子,下调的2个蛋白质分子是补体C3、溶解物载体家族4成员3.这些蛋白质分子涉及细胞分化、细胞迁移、细胞凋亡,以及炎症反应等功能.实验结果提示,瘦素可能通过下丘脑-垂体轴参与免疫调节、雌性生殖及炎症反应等,从而对子宫发挥调节作用.本研究将为加深对瘦素调节子宫作用的认识,并为深入研究女性生殖系统疾病提供新的启示.     

Morphological Changes within the Rat Lateral Ventricle after the Administration of Proteasome Inhibitors

The broad variety of substances that inhibit the action of the ubiquitin-proteasome system (UPS)—known as proteasome inhibitors—have been used extensively in previous studies, and they are currently frequently proposed as a novel form of cancer treatment and as a protective factor in intracerebral hemorrhage treatment. The experimental data on the safest route of proteasome inhibitor administration, their associated side effects, and the possible ways of minimizing these effects have recently become a very important topic. The aim of our present study was to determine the effects of administering of MG-132, lactacystin and epoxomicin, compounds belonging to three different classes of proteasome inhibitors, on the ependymal walls of the lateral ventricle. Observations were made 2 and 8 weeks after the intraventricular administration of the studied substances dissolved in dimethyl sulfoxide (DMSO) into the lateral ventricle of adult Wistar rats. Qualitative and quantitative analysis of brain sections stained with histochemical and inmmunofluorescence techniques showed that the administration of proteasome inhibitors caused a partial occlusion of the injected ventricle in all of the studied animals. The occlusion was due to ependymal cells damage and subsequent ependymal discontinuity, which caused direct contact between the striatum and the lateral nuclei of the septum, mononuclear cell infiltration and the formation of a glial scar between these structures (with the activation of astroglia, microglia and oligodendroglia). Morphologically, the ubiquitin-positive aggregates corresponded to aggresomes, indicating impaired activity of the UPS and the accumulation and aggregation of ubiquitinated proteins that coincided with the occurrence of glial scars. The most significant changes were observed in the wall covering the striatum in animals that were administered epoxomicin, and milder changes were observed in animals administered lactacystin and MG-132. Interestingly, DMSO administration also caused damage to some of the ependymal cells, but the aggresome-like structures were not formed. Our results indicate that all of the studied classes of proteasome inhibitors are detrimental to ependymal cells to some extent, and may cause severe changes in the ventricular system. The safety implications of their usage in therapeutic strategies to attenuate intracerebral hemorrhagic injury and in brain cancer treatment will require further studies.     

Ghrelin对大鼠摄食的影响及orexins信号通路的调控

目的:探究Ghrelin对大鼠摄食的影响及orexins信号通路的调控作用。方法:采用免疫组织化学染色的方法观察Ghrelin免疫阳性神经元轴突末梢与orexin神经元的突触联系以及下丘脑外侧区(LHA)内c-fos的表达。侧脑室注射抗-orexin-A IgG和抗-orexin-B IgG混合液、抗-黑色素浓集激素(MCH)IgG、NPY-1受体拮抗剂后测量大鼠摄食量,观察其对ghrelin诱导摄食的影响。结果:Ghrelin免疫阳性神经元轴突末梢与orexin神经元的突触相接触。侧脑室注射ghrelin可诱导orexin神经元内c-fos表达,但是没有引起MCH神经元内c-fos的表达。预先注射抗-NPY IgG抗体,ghrelin仍然可诱导orexin神经元内c-fos表达。侧脑室预先注射抗-orexin-A IgG和抗-orexin-B IgG抗体可减弱ghrelin促摄食作用,但是预先注射抗-MCH IgG抗体对ghrelin诱导的摄食作用没有明显影响。注射NPY受体拮抗剂可进一步加强抗-orexin-A IgG抗体和抗-orexin-B IgG抗体对ghrelin诱导摄食的抑制效应。结论:ghrelin可能与orexin系统相互作用共同参与摄食和能量平衡的调控。     

侧脑室前角穿刺脑脊液外引流治疗 高血压丘脑出血破入脑39 例临床分析

目的:探讨侧脑室前角穿刺脑脊液外引流治疗高血压丘脑出血破入脑室患者的临床效果。方法:回顾性分析2005年1月至2010年12月应用侧脑室前角单侧或双侧穿刺外脑脊液引流治疗39例高血压丘脑出血破入脑室患者的临床效果(ADL分级)。结果:39例高血压丘脑出血破入脑室患者死亡8例,占20.51%,存活者31例,占79.49%。对存活者随访3个月,按日常生活能力(ADL)分级,ADL1～3级23例,占74.19%,ADL4级7例,占22.58%,ADL5级1例,占3.23%。结论:侧脑室前角穿刺脑脊液外引流术治疗高血压丘脑出血破入脑室者是一种操作简单、损伤小、有效的治疗方法。     

HLRP 蛋白在大鼠脑中的定位及LPS 对其表达的影响

目的:研究HLRP分子在大鼠脑中的细胞定位和表达特点,观察LPS刺激对动物脑HLRP表达的影响。方法:对原代培养的大鼠神经元、星形胶质细胞、小胶质细胞和大鼠脑组织冰冻切片分别进行免疫荧光染色,观察HLRP的细胞定位和表达特点;给大鼠侧脑室注射LPS,提脑组织蛋白,进行Western blot检测,半定量分析LPS刺激后,大鼠脑HLRP的表达变化。结果:①HLRP选择性表达于部分神经元的细胞核中,正常的星形胶质细胞和小胶质细胞不表达HLRP。②从嗅脑到脑干各节段,HLRP在大鼠脑组织中均匀分布,未发现HLRP阳性神经元聚集的现象。③侧脑室注射LPS 1天以后,HLRP表达明显升高(P<0.05)。结论:大鼠脑中正常表达HLRP,侧脑室注射LPS能刺激HLRP表达。     

Ghrelin对改善心衰大鼠左室功能紊乱的作用研究

目的:慢性心力衰竭(CHF)患者终末期阶段常发生左室(LV)重塑和心脏性恶病质,有研究称Ghrelin可能对CHFLV功能和能量代谢产生保护作用。本文旨在探讨Ghrelin对CHF大鼠LV功能紊乱和心源性恶病质的作用。方法:建立左冠状动脉结扎术和假手术组,手术后4周,给予大鼠Ghrelin或生理盐水3周。用超声心动图和心脏导管术监测结果。结果:与给予安慰剂组相比,用Ghrelin治疗的CHF和假手术组,血浆GH和胰岛素样生长因子1明显升高(t=1.49,t=0.71,P0.05)。与Sham-Placebo组相比,CHF-Placebo组大鼠体重明显减轻(t=2.18,P0.05)。然而与CHF-Placebo组相比,CHF-Ghrelin组大鼠,体重(t=3.89,P0.05),心输出量(t=3.28,P0.05),LV dP/dtmax(t=3.90,P0.05)明显增加。Ghrelin增加了CHF大鼠心脏舒张压,抑制LV扩大,增加LV缩短分数。结论:长期注射Ghrelin可改善CHF大鼠LV功能紊乱,减缓LV重构和心脏性恶病质的发展,有望为CHF的治疗提供新的途径和方法。     

Increase of Cerebral Phosphocreatine in Normal Rats after Intracerebroventricular Administration of Creatine

Intracerebroventricular (ICV) administration of creatine increased cerebral phosphocreatine in normal rats by 67%, the highest increase so far reported in an in vivo model. We used osmotic minipumps (Alzet, Palo Alto, CA, USA) to administer creatine, 0.5mM, to the lateral ventricle at the rate of 10 l/h for 3 days. Brain phosphocreatine in saline-treated controls was 33 ± 17 M/g protein (mean ± SD, N = 9). In creatine-treated rats (0.5 mM for 3 days) such content was 55 ± 17 M/g protein (mean ± SD, N = 7). This difference is statistically significant (p = 0.02, t-test). The increase we found in cerebral phosphocreatine is of an order of magnitude comparable to the increase previously found in in vitro experiments, and may be effective in protecting brain tissue from ischemic damage.     

Effect of Continuous Infusion of Anti-L1 Antibody into the Third Cerebral Ventricle above the Suprachiasmatic Nucleus on the Circadian Rhythm of Locomotor Activity in Rats

During an investigation into the role of the neural cell adhesion molecules such as L1 and NCAM in the generation mechanism of circadian rhythms, we observed that L1-like immunoreactive substance is expressed in the hypothalamic suprachiasmatic nucleus (SCN). Therefore, we examined the effect of continuous infusion of anti-L1 antibody into the third cerebral ventricle above the SCN using an Alzet osmotic minipump, on the circadian rhythm of locomotor activity in rats under constant red dim light (less than 1 lx) condition, in order to elucidate the role of L1 in the mechanism of circadian rhythm. Continuous infusion of intact rabbit IgG into the third cerebral ventricle above the SCN, which was done as a control experiment, shifted the phase of the free-running circadian rhythm and reduced daily locomotor activity for an initial few days, however, it did not eliminate the circadian rhythm. In contrast, continuous infusion of anti-L1 antibody temporarily disrupted the circadian rhythm during the infusion period. Furthermore, the infusion of the anti-L1 antibody but not that of control IgG caused a change in the SCN conformation, from which it appeared that SCN neurons displaced in dorsal direction, 4 days after the start of the infusion. These findings suggest that the cell adhesion molecule, L1, might be involved in the generation and/or transduction of the time signal of the circadian rhythm in the SCN.     

Effect of Continuous Infusion of Anti-L1 Antibody into the Third Cerebral Ventricle above the Suprachiasmatic Nucleus on the Circadian Rhythm of Locomotor Activity in Rats

During an investigation into the role of the neural cell adhesion molecules such as L1 and NCAM in the generation mechanism of circadian rhythms, we observed that L1-like immunoreactive substance is expressed in the hypothalamic suprachiasmatic nucleus (SCN). Therefore, we examined the effect of continuous infusion of anti-L1 antibody into the third cerebral ventricle above the SCN using an Alzet osmotic minipump, on the circadian rhythm of locomotor activity in rats under constant red dim light (less than 1 lx) condition, in order to elucidate the role of L1 in the mechanism of circadian rhythm. Continuous infusion of intact rabbit IgG into the third cerebral ventricle above the SCN, which was done as a control experiment, shifted the phase of the free-running circadian rhythm and reduced daily locomotor activity for an initial few days, however, it did not eliminate the circadian rhythm. In contrast, continuous infusion of anti-L1 antibody temporarily disrupted the circadian rhythm during the infusion period. Furthermore, the infusion of the anti-L1 antibody but not that of control IgG caused a change in the SCN conformation, from which it appeared that SCN neurons displaced in dorsal direction, 4 days after the start of the infusion. These findings suggest that the cell adhesion molecule, L1, might be involved in the generation and/or transduction of the time signal of the circadian rhythm in the SCN.