外周免疫刺激诱导小鼠下丘脑室旁核和视上核中Ⅰ型IL-1受体表达的变化

目的　通过研究外周免疫刺激后免疫性细胞因子受体在下丘脑神经内分泌核团中的表达变化 ,揭示这些核团与神经免疫调节的关系。　方法　小鼠腹腔内给予细菌内毒素脂多糖 (lipopolysaccharide ,LPS)或葡萄球菌肠毒素B(staphylococcalenterotoxinB ,SEB) ,用免疫组织化学方法观察脾脏核增殖抗体 (PCNA)及下丘脑室旁核和视上核中Ⅰ型IL 1受体的表达 ,并采用双标记技术观察Ⅰ型IL 1受体阳性神经元和加压素及催产素表达的关系。　结果　 1 与对照组比较 ,LPS或SEB引起脾脏核增殖抗体的表达明显增强。 2 Ⅰ型IL 1受体在正常小鼠脑内的表达很广泛。与对照组比较 ,LPS或SEB引起Ⅰ型IL 1受体在小鼠下丘脑室旁核和视上核中表达显著增强。 3 在正常下丘脑室旁核和视上核中Ⅰ型IL 1受体阳性的神经元既有加压素能的 ,又有催产素能的。　结论　下丘脑室旁核和视上核参与了免疫反应的调节 ,这两个核团中的部分加压素能神经元和催产素能神经元在神经免疫调节中可能具有重要作用     

切除两侧肾上腺对大鼠下丘脑加压素、催产素神经元的影响

为了进一步确定加压素(VP)在应激反应中的作用,我们应用免疫组化和免疫电镜观察了切除两侧肾上腺对大鼠下丘脑室旁核和视上核的神经内分泌细胞的影响,发现下丘脑室旁核VP神经元处于功能亢进状态,其合成、储存和分泌VP的功能均亢进,而视上核的加压素(VP)神经元以及室旁核和视上核的催产素(OT)神经元的功能状态均无明显改变,提示室旁核的VP神经元可能是参与下丘脑—垂体—肾上腺这一应激反应轴调节的重要因素。     

外周免疫刺激诱导小鼠下丘脑室旁核和视上核中I型IL—1受体表达的变化

目的：通过研究外周免疫刺激后免疫性细胞因子受体在下丘脑神经内分泌核团中的表达变化，揭示这些核团与神经免疫调节的关系。方法：小鼠腹腔内给予细菌内毒素脂多糖（lipopolysaccharide,LPS）或葡萄球菌肠毒素B（staphylococcal enterotoxin B,SEB)，用免疫组织化学方法观察脾脏核增殖抗体（PCNA）及下丘脑室旁核和视上核中I型IL－1受体的表达，并采用双标记技术观察I型IL－1受体阳性神经元和加压素及催产素表达的关系。结果：1．与对照组比较，LPS或SEB引起脾脏核增殖抗体的表达明显增强。2．I型IL－1受体在正常小鼠脑内的表达很广泛。与对照组比较，LPS或SEB引起I型IL－1受体在小鼠下丘脑室旁核和视上核中表达显著增强。3．在正常下丘脑室旁核和视上核中I型IL－1受体阳性的神经元既有加压素能的，又有催产素能的。结论：下丘脑室旁核和视上核参与了免疫反应的调节，这两个核团中的部分加压素能神经元和催产素能神经元在神经免疫调节中可能具有重要作用。     

免疫抑制剂雷帕霉素对LPS免疫激发大鼠脑内免疫相关核团神经元Fos和nNOS表达的影响

观察免疫抑制剂雷帕霉素对大鼠中枢免疫核团Fos和nNOS表达的影响,并探讨雷帕霉素对中枢免疫调节的作用。将大鼠分为雷帕霉素(10mg/kg)实验组和生理盐水对照组。两组动物分别给药后,腹腔注射免疫激发剂细菌脂多糖(LPS,25mg/kg),用免疫组织化学方法显示并计数下丘脑室旁核、视上核和孤束核中Fos蛋白阳性、nNOS样免疫阳性神经元和Fos/nNOS双标神经元数量,并对数据进行统计学处理。结果显示:实验组动物的下丘脑室旁核、视上核和孤束核内Fos、Fos/nNOS神经元数目较对照组均显著减少(P<0.01)。这一结果表明免疫抑制剂雷帕霉素降低了中枢免疫相关核团内因LPS免疫应激所引起的nNOS的表达,提示雷帕霉素在抑制机体免疫机能的同时,也降低了神经-内分泌系统对免疫的抑制作用。     

大鼠下丘脑催产素样和加压素样神经元与5—羟色胺及儿苯酚胺能传入纤维…

用免疫组织化学ＡＢＣ双标记法，显示下丘脑各神经分泌核区中催产素样免疫反庆，加压素样免疫反就神经元及５－羟色胺样免疫反应和儿茶酚胺能，即酷胺酸羟化酶免疫瓜纤维末梢，对这两种单胺能纤维末梢在视上核，室旁核及各种神经分泌副核中的分布，与ＯＴ－ｌｉ和ＶＰ－ｌｉ神经元相关的特点及异同加以分析。结果显示，５－ＨＴ－ｌｉ纤维未梢在下丘脑视上核、室旁核及各神经分泌副核中，与ＯＴ－ｌｉ及ＶＰ－ｌｉ神经元相接触，并在     

Fos蛋白在LPS免疫激发大鼠脑内的分布

目的 研究与神经免疫调节有关的功能神经元在大鼠脑内的分布。方法 以腹腔注射脂多糖（ＬＰＳ）为免疫激发模型,采用免疫组织化学ＡＢＣ方法,观察Ｆｏｓ蛋白在脑内的分布情况。结果 Ｆｏｓ阳性产物多集中分布在大脑额顶皮质、边缘前脑（扣带皮质、梨状皮质、外侧隔核和中央杏仁核等）、丘脑室旁核、下丘脑室旁核、弓状核、视上核、视交叉上核、下丘脑外侧区、中脑导水管周围灰质腹外侧部、外侧臂旁核和延髓内脏带,小脑内无明显     

抗利尿激素研究的若干进展

人和大多数哺乳动物的抗利尿激素为精氨酸加压素(arginine vasopressin, AVP)。经典的理论认为,加压素和催产素由视上核和室旁核的大细胞神经元(magnocellular neu-rone)合成,经下丘脑-神经垂体束移行至神经垂体。近年来应用免疫组织化学、电镜等研究的结果表明,室旁核的小细胞加压素神经元(parvocellular vasopressin neurones)及脑干中的某些大细胞神经元也能合成加压素。加压素至少经四个途径被输送到脑的各部     

烫伤或外伤性骨折对下丘脑神经内分泌细胞的影响——Gomori方法、免疫组化和免疫电镜观察

为了从形态学角度进一步探讨VP、OT在应激中的作用,我们利用Gomori染色、免疫组化和免疫电镜观察,发现无论是烫伤或骨折后,大鼠下丘脑室旁核VP神经元的机能状态有明显的改变,合成、储存和分泌VP的功能均亢进,而OT神经元和视上核VP、OT神经元的机能状态无明显改变。我们推测下丘脑室旁核VP神经元的反应是损伤性应激时的变化。     

大鼠下丘脑室旁核和视上核内神经激肽B受体和加压素的共存研究

目的研究神经激肽Ｂ受体（ＮＫ３受体）免疫反应产物在大鼠下丘脑室旁核和视上核的分布及其与加压素的共存。方法免疫组织化学染色,标本在光镜和电镜下观察。共存研究采用相邻切片染色法。结果致密的ＮＫ３受体阳性产物分布于室旁核的后大细胞部和视上核的主部。免疫电镜观察发现ＮＫ３受体阳性反应产物出现于室旁核和视上核的胞体和树突内。在相邻切片上观察到室旁核和视上核内有相当数量的ＮＫ３受体阳性神经元,同时含有加压素,它们主要分布于室旁核的后大细胞部和视上核的主部。结论根据本研究和以往的机能学的研究结果,推测下丘脑室旁核和视上核内的ＮＫ３受体阳性神经元可能参与调控机体内环境改变时下丘脑加压素的释放。     

烫伤后大鼠下丘脑视上核催产素神经元的免疫电镜观察及形态计量分析

用免疫细胞化学的方法结合图像分析技术已经证明烫伤后2小时视上核催产素神经元的总体积显著增长,说明催产素与应激有密切关系.在此基础上,采用免疫电镜方法进一步观察了烫伤后2小时大鼠下丘脑视上核催产素神经元的超微结构变化,并对其进行了形态计量分析.结果表明:烫伤后2小时大鼠下丘脑视上核催产素神经元的粗面内质网、高乐尔基复合体、线粒体的体密度、面密度比对照组显著增大(P<0.01,或P<0.05).提示烫伤后2小时大鼠下丘脑视上核催产素神经元的合成及分泌功能均有所增强.     

Neurohypophyseal peptides in aging and Alzheimer's disease

The neurohypophyseal hormones arginine-vasopressin (AVP) and oxytocin (OT) are produced in the neurons of the hypothalamic supraoptic (SON) and paraventricular (PVN) nucleus and in the much smaller cells of the suprachiasmatic (SCN) nucleus. The SON is the main source of plasma AVP. Part of the AVP and OT neurons of the PVN join the hypothalamo-neurohypophyseal tract, whereas others send projections to the median eminence or various brain areas, where AVP and OT are involved in a number of central functions as neurotransmitters/neuromodulators. AVP and OT from the PVN can also regulate via the autonomous innervation endocrine glands and fat tissue. OT is produced for a major part in the PVN but some OT neurons are present in the SON. Moreover, both AVP and OT containing neurons are observed in the "accessory nuclei", i.e. islands situated between the SON and PVN. The SCN is the biological clock, and the number of AVP expressing neurons in the SCN shows both diurnal and seasonal rhythms. In addition to these hypothalamic areas, AVP and OT may be found to a lesser extent in some other brain areas, such as the bed nucleus of the stria terminalis, diagonal band of Broca, nucleus basalis of Meynert, lateral septal nucleus, globus pallidus and the anterior amygdaloid nucleus, as well as in the peripheral tissues. The AVP and OT containing neurons should not be considered as one system. Prominent functional differences exist between the different nuclei. The heterogeneity also becomes clear from the marked differences in the neurohypophyseal peptides containing neurons of the SON, PVN and SCN during aging, and in the most prevalent age-related neurodegenerative diseases, i.e. Alzheimer's disease (AD). For those reasons, we will discuss the SON, PVN and SCN separately.     

腹腔注射2-脱氧-D-葡萄糖可诱发大鼠视上核和室旁核神经元表达Fos

目的 探讨腹腔注射2-脱氧-D-葡萄糖（2-DG）能否激活大鼠下丘脑视上核（SON）和室旁核（PVN）神经元而表达Fos。方法 健康雄性SD大鼠12只,随机分为腹腔注射2-DG组（6只）、生理盐水对照组（3只）及正常对照组（3只）。各自处理后,应用免疫组织化学方法,观察各组下丘脑SON和PVN内Fos表达及其与催产素（OT）和加压素（VP）的双标情况,同时采用ELISA方法对血清中OT和VP的含量进行检测。 结果与生理盐水对照组和正常对照组相比,2-DG引发的特异性Fos免疫阳性产物主要集中分布于下丘脑外侧区和穹隆周区,在SON、PVN也有密集表达。SON和PVN内的Fos表达与该区的特异性神经活性物质OT和VP有共存。OT/Fos双标细胞率（双标细胞占OT阳性细胞的百分率）在SON和PVN分别为87.10%、90.57%,明显高于VP/Fos在这两个核团的双标率（双标细胞占VP阳性细胞的百分率,68.42%、76.92%）,两者比较差异有统计学意义（P<0.05）。ELISA检测结果显示,2-DG组动物血清中OT和VP水平与对照组相比无明显变化。 结论 腹腔注射2-DG可激活大鼠下丘脑SON和PVN内OT和VP神经元表达Fos,SON和PVN可能参与2-DG诱导的急性应激反应。     

Developmental changes of nitric oxide synthase expression in the rat hypothalamoneurohypophyseal system

The present study investigated the immunohistochemical localization of neuronal nitric oxide synthase (nNOS) in the hypothalamoneurohypophyseal system (HNS) of the developing rats on postnatal day 1 (PN1), 7 (PN7), 14 (PN14), 21 (PN21), and the adult rats. The nNOS-positive neurons were not discernable in the supraoptic nucleus (SON), the paraventricular nucleus (PVN), and the median eminence (ME) at PN1 and PN7. A few neurons positive for nNOS were first detected at PN14. At PN21, the nNOS-positive cells in SON and PVN rapidly increased in number. The pattern of nNOS expression at this stage approached that of the adult. Moreover, the increase of nNOS expression in the SON and PVN during the postnatal period was accompanied by the maturation of arginine vasopressin (AVP) and oxytocin (OT) neurons as indicated by the number and size of OT or AVP neurons in the SON and PVN. The patterns of AVP versus OT expression also reached that of the adult by the end of the third postnatal week. The time course of the change in nNOS expression coincided with the maturation of AVP and OT neurons in the HNS and suggested that NO synthesized by conversion of NOS is involved in the modulation of activity of neurons in the SON and PVN of the HNS.     

大鼠下丘脑催产素及加压素样神经元与脑啡肽样传入纤维末梢的关系

用免疫组织化学ABC双标记法,显示大鼠下丘脑各大细胞神经分泌核团中催产素样免疫反应(OT-li)、加压素样免疫反应(VP-li)神经元、亮-脑啡肽样免疫反应(L-ENK-li)和甲-脑啡肽样免疫反应(M-ENK-li)纤维末梢,并对不同神经分泌核团内脑啡肽样传入纤维末梢与OT-li及VP-li神经元的关系加以分析。结果发现,L-ENK-li及M-ENK-li纤维末梢在下丘脑各神经分泌核团中,与OT-li及VP-li神经元均有一定程度的接触关系。L-ENK-li与M-ENK-li纤维末梢在第三脑室周、前连合核、背内侧和背外侧副核、穹窿前和后核的OT-li及VP-li神经元周围最密集,在视上核、室旁核次之,在血管周细胞群内的OT-li及VP-li神经元周围密度较低。这些神经分泌性核团内L-ENK-li纤维末梢,均较M-ENK-li纤维末梢密集。表明脑啡肽能传入纤维末梢,与视上核、室旁核及各神经分泌副核(除血管周细胞群外)中的OT-li及VP-li神经元,均有不同程度的接触关系。因此,脑啡肽能传入纤维末梢可能在大细胞神经分泌系统OT-li及VP-li神经元分泌活动的调节中,起较广泛的重要作用。     

Colocalization of oxytocin and phosphorylated form of elongation factor 2 in the rat hypothalamus

Oxytocin (OT) is one of the neuropituitary hormones and is synthesized in the neurons of the paraventricular nucleus (PVN) and supraoptic nucleus (SON). Previous studies have shown that the mRNAs encoding OT are delivered from the soma to both dendrites and axons of the neurons in the PVN and SON. However, it has not been elucidated whether a translational regulation mechanism to enable local synthesis of the hormone exists in the axons of the neurons of PVN and SON. Elongation factor 2 (EF2) is essential for polypeptide synthesis during protein translation. Moreover, phosphorylation of EF2 by EF2 kinase enhances the translation of certain mRNA species. In the present study, in order to shed light on the mechanisms involved in the translational regulation of OT synthesis, we investigated the localization of phosphorylated EF2. Phospho-EF2 was localized in the soma of the neurons in PVN and SON, and in the swellings of the median eminence where axonal tracts of the neurons in the PVN and SON exist. The phosphorylated form was also observed in the rat hypophysis. Moreover, phospho-EF2 and OT were colocalized in a part of the neurons in the PVN and SON. These results suggest that OT may be partially translated in the axons of neurons in the PVN and SON, and then secreted from the pituitary.     

Chronic morphine treatment inhibits oxytocin release from the supraoptic nucleus slices of rats

Effect of chronic morphine treatment on oxytocin (OT) release from the long term-cultured organotypic slice of the supraoptic nucleus (SON) was investigated using radioimmunoassay. The co-localization of oxytocin and mu-opioid receptor in neurons within the SON was observed with the double-labeled methods of in situ hybridization combined with immunohistochemistry. After exposure to morphine for 6days, the OT levels in culture media were significantly decreased. Naloxone caused much greater release of OT in chronic morphine treatment group than in controls. Naloxone has no effect after acute morphine treatment. 90% of OT-ir (immunoreactive) neurons expressed mu-opioid receptor mRNA in the SON and 45% of the neurons that expressed mu-opioid receptor mRNAs were OT-ir neurons. These results indicated that the neurons within SON could develop dependence on morphine in vitro, and these effects might be exerted via mu-opioid receptor in oxytocin neurons of the SON.     

大鼠下丘脑催产素样和加压素样神经元与5-羟色胺及儿茶酚胺能传入纤维末梢的关系

用免疫组织化学ABC双标记法,显示下丘脑各神经分泌核区中催产素样免疫反应(oxytocin-like immunoreactivity,OT-li)、加压素样免疫反应(vasopressin-like immunoreaetivity,VP-li神经元及5-羟色胺样免疫反应(serotonin-like immunoreactivity,5-HT-li)和儿茶酚胺能,即酪胺酸羟化酶免疫反应(tyrosine hydroxylase-like immunoractivity,TH-li)纤维末梢,对这两种单胺能纤维末梢在视上核、室旁核及各种神经分泌副核中的分布,及其与OT-li和VP-li神经元相关的特点及异同加以分析。结果显示,5-HT-li与TH-li纤维末梢在下丘脑视上核、室旁核及各神经分泌副核中,与OT-li及VP-li神经元相接触的多寡程度多不相同。视上核、室旁核内有极高密度的TH-li纤维末梢,与OT-li及VP-li神经元相接触,并在多数神经分泌副核的OT-li及Vp-li神经元周围,也有多寡不等的分布;而中等至高密度的5-HT-li纤维末梢,主要与各神经分泌副核内的OT-li或VP-li神经元相接触。说明5-HT-li和儿茶酚胺能传入纤维末梢,可能分别侧重调控不同神经分泌核区的大细胞神经分泌细胞群。     

大鼠下丘脑弓状核神经元的神经支配

用ＨＲＰ逆行追踪和免疫细胞化学与电镜结合法研究了下丘脑弓状核神经元的神经支配。结果表明，室旁核向弓状核投射的神经元中，有一部分为催产素免疫反应阳性，一部分为后叶加压素免疫反应阳性，视上核也有部分ＯＴ免疫反应阳性神经元发出纤维投射至弓状核。将零乱毒素Ｂ亚单位结合ＨＲＰ注入第三脑室后，可见弓状核内有逆行标记细胞，电镜观察发现弓状核内的脑啡肽免疫反应阳性的树突接受ＨＲＰ反应阳性的触液神经元的轴突形成突触     

加压素能和宫缩素能神经元在新生儿下丘脑的分布

本文用ABC法对加压素样和宫缩素样神经元在新生儿下丘脑的分布进行了观察。结果发现,两者的神经元集中分布于视上核、室旁核及副大细胞分泌核。根据两者的分布特点,又将视上核、室旁核和副大细胞分泌核再分为若干分区,各分区均以加压素样神经元为主,约占总数的80～90％。本文结果提示加压素样和宫缩素样神经元在下丘脑的分布具有动物种属和年龄特征。