GnRH受体和5-HT受体与更年期大鼠潮热的关系

目的：研究促性腺激素释放激素(GnRH)受体、5-羟色胺(5-HT)受体和更年期潮热的关系,为诊治潮热提供思路。方法：手术去卵巢后建立SD大鼠更年期潮热的动物模型,采用免疫组织化学邻片法、HE染色及图像分析等方法来检测大鼠垂体中2种受体的表达。结果：大鼠去卵巢后第8w开始,实验组动物肛温明显高于对照组,垂体中实验组GnRH受体和5-HT受体的表达,明显强于对照组。结论：雌激素缺乏可引起类似于更年期潮热的体温升高;在潮热的发病机制中,GnRH和5-HT可能通过其受体发挥重要作用。     

妊娠与非妊娠大鼠子宫内膜促性腺激素释放激素及其受体的免疫组织化学研究

目的：研究促性腺激素释放激素（GnRH)及其受体在妊娠与非妊娠大鼠子宫内膜中的分布及相对含量,为了解促性腺激素释放激素对子宫内膜的可能功能提供依据。方法：免疫组织化学ABC法及图像分析技术。结果：非妊娠大鼠子宫内促性腺激素释放激素及促性腺激素释放激素受体阳性反应发生在内膜上皮、腺上皮及基质细胞。妊娠大鼠子宫内这二者阳性反应主要发生在内膜上皮及基蜕膜的蜕膜细胞,妊娠大鼠子宫上皮细胞中的促性腺激素释放激素及其受体的相对含量明显高于非妊娠大鼠子宫内膜。结论：妊娠与非妊娠大鼠子宫内膜均能表达促性腺激素释放激素及其受体。     

GnRH激动剂主动免疫对GnRHR在腺垂体与子宫表达及分布的作用研究

目的探讨GnRH激动剂主动免疫对绵羊腺垂体与子宫GnRHR表达及分布的影响,为深入研究GnRH-A调节生殖功能的机理及合理应用提供依据。方法 28只5～6月龄健康母绵羊(Ovis aries)随机分为4组(n=7),实验Ⅰ组(EG-Ⅰ)、实验Ⅱ组(EG-Ⅱ)和实验Ⅲ组(EG-Ⅲ)于0 d和14 d分别皮下注射阿拉瑞林抗原200μg、300μg和400μg(0 d和14 d各1次);对照组(CG)皮下注射2.0 ml药物的溶媒(0 d和14 d各1次)。各组于70 d无菌切取腺垂体和子宫。提取腺垂体总RNA,实时荧光定量PCR(FQ-PCR)检测GnRHR mRNA表达的变化,Western blotting分析子宫GnRHR蛋白表达,免疫组织化学SP法染色检测GnRHR表达的变化。结果 EG-Ⅰ、EG-Ⅱ和EG-Ⅲ腺垂体GnRHR mRNA表达量均低于对照组,以EG-Ⅲ最小(P<0.01)。与CG相比,EG-Ⅰ、EG-Ⅱ和EG-Ⅲ子宫GnRHR蛋白表达水平分别减少3.46%、4.90%和24.78%(P<0.05)。子宫组织中有GnRHR分布主要见于子宫内膜细胞和子宫腺上皮细胞的胞质和胞核,EG-Ⅲ灰度值显著低于CG(P<0.05)。结论 GnRH激动剂主动免疫可以剂量依赖性地抑制垂体GnRHR mRNA和子宫组织中GnRHR蛋白的表达。GnRHR主要分布在子宫内膜上皮细胞和腺上皮细胞的胞核和胞质,GnRHR激动剂免疫对子宫中GnRHR的分布具有抑制作用。     

人甲状腺促性腺激素释放激素及其受体的表达

目的　探讨人甲状腺中是否存在促性腺激素释放激素 (GnRH)及促性腺激素释放激素受体 (GnRH R)并细胞定位 ,试图从转录及翻译水平讨论人甲状腺可否合成GnRH及GnRH R ,为探讨GnRH可能影响甲状腺的功能提供形态学依据。　方法　采用免疫组织化学法和原位杂交技术。　结果　所测人正常甲状腺组织均呈较强的GnRH和GnRH R免疫反应阳性 ,甲状腺滤泡上皮细胞、滤泡旁细胞均为阳性细胞。免疫反应阳性物质主要分布在阳性细胞胞质内 ,胞核呈阴性。原位杂交结果同免疫组织化学染色基本一致 :甲状腺滤泡上皮细胞胞质和滤泡旁细胞胞质可检测到较强的GnRHmRNA及GnRH RmRNA阳性杂交信号。胞核均呈阴性 ,未检测到杂交信号。　结论　人甲状腺不仅表达GnRH和GnRH R ,而且可以自身合成GnRH和GnRH R。GnRH可能以自分泌的方式影响甲状腺的生理功能     

人胎胰腺GnRH免疫反应细胞

目的：探讨促性腺激素释的激素(GnRH)免疫反应细胞在人胎胰腺的存在部位和数量变化。方法：用免疫组织化学SABC法,对37例第10-32w人胎胰腺内的GnRH-IR细胞进行观察,并用体视方法分析其数量变化。结果：人胎胰腺GnRH-IR细胞出现于第13w,其数密度随胎龄增加而增大;分布于胰岛及外分泌部的腺泡上皮、导管上皮细胞间。位于胰岛的GnRH-IR细胞呈圆形、卵圆形或多边形。位于腺泡上皮细胞间的GnRH-IR细胞多为锥体形,外分泌部的GnRH-IR细胞均为开放型细胞。结论：胰腺GnRH-IR细胞于胚胎第13w出现,广泛存在于内、外分泌部,其数量随胎龄增加而增加。     

GnRH拮抗剂与GnRH激动剂短方案IVF-ET结局的比较

目的比较GnRH antagonist与GnR Hagonist短方案的IVF-ET结局。方法2006年8月至2007年8月GnR Hantagonist治疗组54人和GnR Hagonist短方案对照组132人,记录促性腺激素的用量及其用药天数、hCG日子宫内膜厚度和激素水平、获卵数、受精率、卵裂率、优胚率、妊娠率和OHSS发生率等指标。结果两组促性腺激素的用量及其用药天数、获卵数、受精率、卵裂率、着床率和妊娠率相比较均无显著差异（P〉0.05）。GnR Hantagonist组在hCG日激素水平低,与对照组比较其差异有统计学意义。结论行GnR Hantagonist方案IVF-ET助孕治疗与传统的GnR Hagonist短方案比较,其hCG日雌激素水平下降可能是OHSS发生率显著下降的主要因素;但卵泡的发育、卵母细胞的受精率、卵裂率及妊娠率和着床率均不受影响。GnR Hantagonist的使用为IVF-ET助孕药物提供了一种新的选择。     

大鼠下颌下腺卵泡刺激素受体、雌激素α受体的分布及与促性腺激素释放激素受体的共存

目的 研究卵泡刺激素受体( FSHR)和雌激素α受体(ERα)在大鼠下颌下腺中的定位、分布及与促性腺激素释放激素受体( GnRHR)共存.方法采用免疫组织化学定位和邻片免疫组织化学共定位方法.结果大鼠下颌下腺浆液性腺泡及各级导管的上皮细胞均呈FSHR和ERα免疫反应阳性;在下颌下腺GnRHR免疫反应阳性细胞中观察到了F...     

促性腺激素释放激素受体在雌性山羊颈胸神经节中的分布

目的探讨颈胸神经节是否具备接受促性腺激素释放激素(GnRH)作用的条件。方法实验采用免疫组织化学SP法,观察GnRH受体在雌性山羊颈胸神经节中的分布特点。结果在颈胸神经节中,神经元胞体均为GnRH受体免疫反应阳性,其细胞膜以及膜下胞质中的颗粒状物质呈棕黄色,GnRH受体为强阳性表达,而胞核呈阴性表达。部分神经元可见其突起,呈黄色,GnRH受体为中等阳性表达。卫星细胞也呈黄色,GnRH受体为中等阳性表达。图像分析表明,呈阳性反应的神经元胞体与节内其他阳性成分相比,其GnRH受体相对表达量差异性极显著(P0.01)。结论在雌性山羊颈胸神经节中,神经元胞体是GnRH发挥作用的主要靶细胞,颈胸神经节具备接受GnRH作用的条件。提示GnRH有可能通过作用于颈胸神经节交感节后神经元上的GnRH受体来影响心肺器官的功能活动,即颈胸神经节可能是心肺功能活动中协调GnRH内分泌调节和该神经节自主神经调节两途径之间的中继站。     

GnRH拮抗剂在体外受精-胚胎移植中的应用

目的探讨促性腺激素释放激素（GnRH）拮抗剂方案在体外受精-胚胎移植（IVF—ET）促超排卵中的应用效果。方法回顾性比较分析本中心2006年8月～2007年8月接受ⅣF—ET助孕治疗的患者中采用GnRH拮抗剂方案的54例患者和采用GnRH激动剂长方案的135例患者，观察其临床效果。结果两组Gn用量、HCG日内膜厚度、受精率、卵裂率之间比较无显著性差异；两组患者Gn使用天数、HCG日E2值、获卵数、冷冻率、种植率、妊娠率、OHSS发生率之间比较有显著性差异。结论GnRH拮抗剂联合促性腺激素促超排卵方案缩短用药时间，减少费用，并可显著降低OHSS的发生率，但冷冻率、妊娠率较GnRH激动剂长方案偏低。     

乳腺浸润性导管癌中促性腺激素释放激素受体的表达及意义

目的 探讨促性腺激素释放激素受体(gonadotropin-releasing hormone,GnRHR)在乳腺浸润性导管癌中的表达及意义.方法 采用免疫组化方法 检测50例乳腺浸润性导管癌及18例正常乳腺组织中的GnRHR.结果 乳腺浸润性导管癌中GnRHR的阳性表达为68%(34/50).其中乳腺癌 G1组的阳性表达(92.3%,12/13)与G3组的阳性表达(46.2%,6/13)之间的差异有统计学意义(P<0.05).乳腺癌G3组的阳性表达和正常乳腺组织的阳性表达(94.4%,17/18)之间的差异有统计学意义(P<0.01).结论 大多数正常乳腺组织的GnRHR虽有表达,但呈弱阳性表达,而多数浸润性导管癌GnRHR则呈强阳性表达,提示乳腺组织的分化程度影响肿瘤组织内GnRHR的表达.     

GnRH-A促进雌兔FSH与LH分泌及卵巢发育的作用

目的探讨GnRH类似物(GnRH-A)对FSH与LH分泌及卵巢发育的影响,及GnRH-A调节生育的机制。方法 24只雌兔均分为实验Ⅰ组(EG-Ⅰ)、Ⅱ组(EG-Ⅱ)、Ⅲ组(EG-Ⅲ)和对照组(CG)(n=6),实验组分别于颈背侧皮下注射0.1、0.1和0.05 g/L GnRH-A抗原1.0 mL,EG-Ⅱ和EG-Ⅲ组于20 d加强注射1次。ELISA法测定血清FSH和LH含量。于70 d无菌切取卵巢,光镜和电镜观察。结果 40 d时EG-Ⅱ和EG-Ⅲ组血清FSH和LH浓度均达到峰值,明显高于EG-Ⅰ和CG组的FSH和LH(P<0.01),且EG-Ⅱ组高于EG-Ⅲ组(P<0.05)。EG-Ⅱ组和EG-Ⅲ组卵巢皮质变厚,卵泡纵径和横径均增大,且与剂量相关。实验组卵巢卵泡数增加,生长加快。细胞核、线粒体和线粒体嵴变大,细胞质中的皮质颗粒和分泌物增多,透明带和微绒毛变大。结论 GnRH-A能促进FSH和LH的合成及卵巢与卵泡的发育,加强注射效果更明显。     

GnRH激动剂主动免疫母羊对生殖激素分泌的作用

目的:探讨GnRH激动剂(GnRHa)主动免疫对绵羊生殖激素合成与分泌的作用,并深入研究GnRH-A免疫调节动物生殖功能的机理。方法:42只5～6月龄母绵羊(Ovis aries)随机分为6组(n=7),EG-Ⅰ、EG-Ⅱ和EG-Ⅲ分别于0和14天皮下注射阿拉瑞林抗原200、300和400μg;EG-Ⅳ和EG-Ⅴ分别皮下注射阿拉瑞林抗原200、300、0、7、14和21天各一次,共4次;对照组在0和14天皮下注射抗原溶媒(除不用阿拉瑞林外,其余成分和制备方法与阿拉瑞林抗原相同)2.0 ml。无菌采集不同时段的血液,分离血清。以ELISA测定血清GnRH抗体浓度,用激素检测试剂盒(ELISA)分别测定血清GnRH、FSH、LH和E2浓度。结果:①阿拉瑞林首次免疫7天后,各实验组的抗体浓度逐渐升高,EG-Ⅰ、EG-Ⅱ和EG-Ⅲ分别在28、28和35天达到峰值(P<0.05),EG-IV和EG-V则在在45天达到峰值(P<0.01),至60天时仍明显高于对照组(P<0.05)。14～60天间EG-IV和EG-V抗体浓度均高于EG-Ⅰ、EG-Ⅱ和EG-Ⅲ(P<0.05)。②EG-Ⅰ、EG-Ⅱ的GnRH在21和28天抵谷值(P<0.05),EG-Ⅲ、EG-Ⅳ和EG-Ⅴ则在45天抵谷值(P<0.01),且以EG-Ⅴ为最低。谷值之后逐渐上升趋势,70天时达到免疫注射前水平。③实验组血清FSH浓度始终高于对照组(P<0.05)。EG-Ⅰ、EG-Ⅱ和EG-Ⅲ于28、28和35天达到峰值(P<0.05),而EG-IV和EG-V在60天达到高峰值(P<0.01)。④实验组绵羊血清LH呈下降趋势,EG-Ⅰ、EG-Ⅱ和EG-Ⅲ分别在21、21和28天达到谷值(P<0.01),EG-Ⅳ和EG-Ⅴ在35天达到谷值(P<0.01)。35天时EG-Ⅳ和EG-Ⅴ低于EG-Ⅰ、EG-Ⅱ和EG-Ⅲ。⑤各组的血清E2含量无显著差异。结论:GnRH激动剂(阿拉瑞林)抗原主动免疫可促进GnRH抗体的生成,抑制母羊GnRH和LH的合成与分泌,增强FSH的合成与分泌,且随着注射剂量和注射次数的增加,这种作用更加明显,而对血清E2无明显影响。     

免疫荧光法检测FSHR与GnRHR在胃癌组织中的分布及其共定位研究

研究卵泡刺激素受体与促性腺激素释放激素受体在胃癌组织中的定位、分布及共存性。方法 选取我院胃肠外科经手术和病理证实的48例胃癌患者的病理组织石蜡包块标本,采用免疫荧光双标记定位方法检测胃癌标本中FSHR及GnRHR的定位及分布。结果 FSHR和GnRHR在胃癌组织细胞胞浆中均有分布,免疫荧光反应阳性物质分布于细胞质,细胞核呈阴性反应,二者分布模式相同。结论 FSHR与GnRHR在胃癌组织中分布具有共存性,提示其可能对胃癌的病理发生机制具有重要的影响。     

The existence of gonadotropin-releasing hormone-like peptides in the neural ganglia and ovary of the abalone, Haliotis asinina L.

Gonadotropin-releasing hormone (GnRH) is a neuropeptide that is conserved in both vertebrate and invertebrate species. In this study, we have demonstrated the presence and distribution of two isoforms of GnRH-like peptides in neural ganglia and ovary of reproductively mature female abalone, Haliotis asinina, using immunohistochemistry. We found significant immunoreactivities (ir) of anti-lamprey(l) GnRH-III and anti-tunicate(t) GnRH, but with variation of labeling intensity by each anti-GnRH type. lGnRH-III-ir was detected in numerous type 1 neurosecretory cells (NS1) throughout the cerebral and pleuropedal ganglia, whereas tGnRH-I-ir was detected in only a few NS1 cells in the dorsal region of cerebral and pleuropedal ganglia. In addition, a small number of type 2 neurosecretory cells (NS2) in cerebral ganglion showed lGnRH-III-ir. Long nerve fibers in the neuropil of ventral regions of the cerebral and pluropedal ganglia showed strong tGnRH-I-ir. In the ovary, lGnRH-III-ir was found primarily in oogonia and stage I oocytes, whereas tGnRH-ir was observed in stage I oocytes and some stage II oocytes. These results indicate that GnRH produced in neural ganglia may act in neural signaling. Alternatively, GnRH may also be synthesized locally in the ovary where it could induce oocyte development.     

GnRHa主动免疫对幼鼠子宫发育的作用

目的探讨促性腺激素释放激素激动剂(GnRHa)主动免疫对幼鼠子宫发育的作用。方法 60只昆明雌鼠随机均分为四组,分别颈部皮下注射不同剂量阿拉瑞林抗原,各组均连续注射7 d。在0 d、7 d、14 d和21 d测定体重,于21 d处理小鼠,显微镜观察子宫的组织结构变化,并用Motic imagles软件测定分析图像数据。结果阿拉瑞林能明显抑制子宫的发育,且剂量越大作用越明显。EG-Ⅲ的UWT明显缩小(P<0.05);实验组EET均小于对照组(P<0.05)。EG-Ⅰ子宫腔轻度缩小;EG-Ⅱ子宫腔和腺体管腔缩小,子宫管壁明显变薄;内膜皱襞减少,上皮变薄;EG-Ⅲ子宫壁变薄,子宫腺减少,内膜细胞胞核变小,上皮变薄,胞质明显减少。结论阿拉瑞林主动免疫能显著抑制幼鼠的子宫发育,且连续重复免疫对小鼠具有毒性作用,剂量越大,作用越明显。     

Detection of gonadotropin-releasing hormone receptor in normal human pituitary cells and pituitary adenomas using immunohistochemistry

Gonadotropin-releasing hormone (GnRH), which is a well-known regulator of gonadotroph function, has recently been considered to be a paracrine factor involved in the control of somatotroph, lactotroph, and corticotroph cells. GnRH action is initiated by binding to a specific cell surface receptor, the gonadotropin-releasing hormone receptor (GnRHR), which is expressed by follicle-stimulating hormone/luteinizing hormone (FSH/LH) cells. Using in situ hybridization techniques, GnRHR messenger ribonucleic acid (mRNA) has recently been detected in normal human anterior pituitary gland and in various pituitary adenomas, including FSH/LH-cell, growth hormone (GH)-cell, adrenocorticotropic hormone (ACTH)-cell, and null-cell adenomas. However, immunohistochemical studies indicating the specific cell distribution of GnRHR in normal pituitary cells have never been reported. The aim of the present investigation was to evaluate the immunohistochemical expression of GnRHR in different types of normal pituitary cells and related tumors. Using double-label immunohistochemical techniques on formalin-fixed and paraffin-embedded tissues and specific antibodies directed against pituitary hormones and GnRHR, we found GnRHR immunoreactivity not only in FSH/LH cells, but also in GH- and thyroid-stimulating hormone (TSH) cells. GnRHR was detected in FSH/LH-cell, GH-cell, mixed GH- and prolactin (PRL)-cell, and α-subunit (α-SU)/null-cell adenomas. The findings of this study suggest that the interaction between GnRH and GnRHR may play a role in paracrine/autocrine regulation of different types of normal pituitary cells and pituitary adenomas. Received: 24 January 2000 / Accepted: 12 April 2000     

Comparison of the Inhibitory Effect of Gonadotropin Releasing Hormone (GnRH) Agonist,Selective Estrogen Receptor Modulator (SERM), Antiprogesterone on Myoma Cell Proliferation In Vitro

Uterine myomas are the most common gynecologic tumor in women of reproductive age. Treatment options of uterine myomas consist of surgical, medical and interventional therapy such as uterine artery embolization or myolysis. Given that it is the most common type of tumor in women of reproductive age, the treatment of uterine myomas must prioritize uterine conservation. There are several drugs for medical treatment of uterine myoma such as gonadotropin releasing hormone (GnRH) agonist, selective estrogen receptor modulator (SERM) and antiprogesterone. The objective of this study was to compare the effect of GnRH agonist, SERM, and antiprogesterone in the treatment of uterine myomas in vitro. The effect of drugs was evaluated through the cell viability assay in cultured leiomyoma cells, western blot analysis of proliferating cell nuclear antigen (PCNA), and BCL-2 protein expression. As a result, mifepristone single-treated group represents the most significant reduction in myoma cell viability and proliferation. When pretreated with leuprolide acetate, raloxifene shows more significant reduction in myoma cell viability and proliferation than mifepristone. This study suggests one of the possible mechanisms how medications act on uterine myoma, especially at the molecular level.     

Distribution and origin of secretoneurin-immunoreactive nerves in the female rat uterus

Secretoneurin is a 33-amino acid peptide derived from secretogranin II. Secretoneurin immunoreactivity has been localized in the peripheral nervous system where it exerts potent chemotactic activity for monocytes and may play a role in inflammation. Secretoneurin could play a role in this process, although the presence and distribution of secretoneurin-immunoreactive neurons in the female reproductive system has not been documented. Thus, this study was undertaken to examine secretoneurin immunoreactivity in nerves of the rat uterus and uterine cervix. A moderate plexus of secretoneurin-immunoreactive nerve fibers was present in the myometrium and endometrium of the uterus as well as in the smooth muscle and endocervix of the cervix. Many of these fibers were associated with the vasculature as well as the myometrium. Secretoneurin immunoreactivity was present in small- to medium-sized neurons of dorsal root and nodose ganglia. Retrograde tracing with FluoroGold indicated that some of these sensory neurons project axons to the cervix and uterine horns. Secretoneurin-immunoreactive terminal-like structures were associated with neurons in the sacral parasympathetic nucleus of the lumbosacral spinal cord. In addition, some secretoneurin terminals were apposed to pelvic parasympathetic neurons in the paracervical ganglia that projected axons to the uterus and cervix. Double-immunostaining indicated co-existence of calcitonin gene-related peptide or substance P with secretoneurin in some sensory neurons, in some terminals of the pelvic ganglia, as well as nerve fibers in the uterine horn and cervix. Finally, fibers in the uterus and cervix were depleted of secretoneurin by capsaicin treatment. This study indicates that secretoneurin is present in the uterus in C-afferent nerve fibers whose cell bodies are located in sensory ganglia. Some of these fibers contain both secretoneurin and calcitonin gene-related peptide or substance P. These substances have functions in inflammatory reactions. Further, secretoneurin could influence postganglionic parasympathetic "uterine-related" neurons in the pelvic ganglia and preganglionic parasympathetic neurons in the lumbosacral spinal cord.