Gonadotrophin-releasing hormone-Ⅰ and -Ⅱ stimulate steroidogenesis in prepubertal murine Leydig cells in vitro

Aim： To study the effect and mechanism of gonadotrophin-releasing hormone （GnRH） on murine Leydig cell steroidogenesis. Methods： Purified murine Leydig cells were treated with GnRH-Ⅰ and -Ⅱ agonists, and testosterone production and steroidogenic enzyme expressions were determined. Results： GnRH-Ⅰ and -Ⅱ agonists significantly stimulated murine Leydig cell steroidogenesis 60%-80% in a dose- and time-dependent manner （P 〈 0.05）. The mRNA expressions of steroidogenic acute regulatory （STAR） protein, P450scc, 3β-hydroxysteroid dehydrogenase （HSD）, but not 17β-hydroxylase or 17β-HSD, were significantly stimulated by both GnRH agonists with a 1.5- to 3-fold increase （P 〈 0.05）. However, only 3β-HSD protein expression was induced by both GnRH agonists, with a 1.6- to 2-fold increase （P 〈 0.05）. Conclusion： GnRH directly stimulated murine Leydig cell steroidogenesis by activating 3β-HSD enzyme expression.

Exogenous gonadotropin releasing hormone (GnRH) modulates scrotal and testicular biometrics, libido, endocrinological and heamatological profiles in Ganjam goat under humid tropical coastal ecosystem of Odisha

Objective:To assess the effect of exogenous gonadotropin releasing hormone(GnRH)in libido,scrotal and testicular biometrics,endocrinological and heamatological profiles of Ganjam goat buck during winter and pre-monsoon seasons.Methods:Forty eight healty Ganjam goat bucks of 3-4 years old were equally divided into the control and treatment groups.The control group received distilled water as placebo,while the treatment group received GnRH injection(4μg Buserelin acetate/mL)once a week for four months.Body weight,scrotal circumference and testicular parameters(testicular volume,testicular weight and testis index),endocrinological profiles[follicle stimulating hormone(FSH),luteinizing hormone(LH),and testosterone],sex behavioural profiles(libido score,mating ability score,and sex behavioural score)and heamatological profiles(red blood cells,white blood cells,haemoglobin,and packed cell volume)were estimated.Results:The libido,scrotal and testicular biometrics,and endocrinological profiles significantly differed between the control and GnRH-treated bucks within and between the seasons within the two groups(P<0.05).Body weight,scrotal circumference,testicular volume,testicular weight,testis index,FSH,LH,testosterone,libido score,mating ability score,sex behavioural score,red blood cells,haemoglobin and packed cell volume were significantly higher in the GnRH-treated bucks compared to the control bucks in pre-monsoon and winter seasons(P<0.05).Body weight,scrotal circumference,testicular volume,testicular weight,testis index,FSH,LH,testosterone,libido score,mating ability score,sex behavioural score,red blood cell counts(in treatment),and haemoglobin were significantly higher in pre-monsoon compared to winter season in the experimental groups(P<0.05).The white blood cell counts neither differed between seasons nor between the two groups.Conclusions:Exogenous GnRH supplementation and pre-monsoon season have higher beneficial effects in improvement of endocrinological profiles with cascading beneficial effects on scrotal circumference,testicular volume,testicular weight,and sex behavioural profiles,which in turn will improve the sperm production and its cryo-survivability and fertility rate in Ganjam goat.

Anti-Müllerian hormone and antral follicle count predict ovarian response in women less than 45 years following GnRH antagonist multiple-dose protocol

Objective:To speculate which of the following parameters:antral follicle count(AFC),anti-Müllerian hormone(AMH),follicle-stimulating hormone(FSH)and age can be used as a predictor of ovarian response to gonadotropin-releasing hormone(GnRH)antagonist stimulation multiple-dose protocol in women under 45 years,and to determine the cutoff value of these parameters and their correlations for predicting low and high ovarian response.Methods:This prospective study included 462 women with the mean age of(29.3±6.5)years.All women were subjected to the GnRH antagonist stimulation multiple-dose protocol.On the second day of the menstrual cycle,ultrasonography was conducted to determine AFC in both ovaries.Peripheral blood samples were collected to evaluate the level of estradiol,FSH,luteinizing hormone,prolactin,thyroid-stimulating hormone,and AMH.The women were divided into three groups:low response(AHH<1 ng/mL,n=173),normal response(AMH=1.0-3.5 ng/mL,n=175),and high response(AMH>3.5 ng/mL,n=114).Results:A significant decrease was found in the age and FSH level in the high response group compared to other groups(P<0.001).Conversely,a significant increase was shown in AMH,estradiol on human chorionic gonadotropin(hCG)day,AFC,mature oocytes,fertilized oocytes,and embryos transferred in the high response group compared to the other two groups(P<0.001).The receiver operating characteristic(ROC)curves demonstrated that AFC and AMH had the highest accuracy,followed by basal FSH level and age in the prediction of low ovarian reserves(P<0.001)with cutoff values of≤4.50 and≤0.95 for AFC and AMH,respectively.Moreover,the ROC analysis showed that AFC had the highest accuracy,followed by AMH level and age in the prediction of high ovarian reserves with a cutoff value of≥14.50,≥3.63,and≤27.50 years,respectively(P<0.01).A significant decrease was observed in women's age,estradiol level,and oocyte fertilization rate in pregnant women compared to non-pregnant women(P<0.001).Additionally,significant negative correlations were found between the AFC,the number of mature oocytes,fertilized oocytes,embryos transferred,and the age of pregnant women(P<0.001).Conclusions:AFC and AMH predict low and high ovarian response to GnRH antagonist stimulation multiple-dose protocol in women under 45 years.

The effect of Wujiadan Recipe on serum sex hormone and expression of Kiss-1, GPR54 and GnRH in arcuate nucleus of hypothalamus in ovariectomized rats

Objective:To explore the effect of wujiadan Recipe on the expression of FSH,E2 in serum and KiSS-1,GPR54,GnRH mRNA in hypothalamic arcuate nucleus tissue homogenate of ovariectomized rats,and to further explore the mechanism of wujiadan Recipe on menopausal syndrome.Methods:Sixty-four adult female rats were randomly divided into three groups:blank group,model group,Chinese medicine control group,western medicine control group,and wujiadan high,medium and low dose groups.Except for the blank group,all the other groups were ovariectomized.The rats in the normal group and the model group were given the same dose of saline once a day.One rat in the blank group died during the experiment.After 4 weeks of continuous administration,blood was collected from abdominal aorta and brain stereotaxic localization was performed to preserve the arcuate nucleus of hypothalamus.The levels of FSH and E2 in serum were detected by enzyme-linked immunosorbent assay(ELISA),and the expression of Kiss-1,GPR54 and GnRH mRNA in arcuate nucleus of hypothalamus were detected by PCR.Results:1.Compared with the blank group,FSH level in the model group was significantly higher(P<0.01),and E2 level was significantly lower(P<0.01).Compared with the model group,the level of serum E2 in the western medicine control group increased significantly(P.The serum FSH content in each group decreased,especially in the high dose group and the western medicine control group(P<0.01),and in the Chinese medicine control group,the middle dose group and the low dose group(P<0.05).Compared with the blank group,the expression of KiSS-1 mRNA,GPR54 mRNA and GnRH mRNA in the model group increased significantly(P<0.01);compared with the model group,the expression of KiSS-1 mRNA and GPR54 mRNA in the high dose group,middle dose group and Western medicine control group of wujiadan prescription decreased significantly(P<0.05).Compared with the model group,the expression of GnRH mRNA in the high dose group and the western medicine control group decreased significantly(P<0.01);the middle dose group of wujiadan decreased significantly(P<0.05).Conclusion:Wujiadan recipe can improve the serum sex hormone level of castrated rats,and reduce the over expression of KiSS-1,GPR54 and GnRH mRNA in hypothalamic arcuate nucleus of castrated rats,which may be the reproductive endocrine mechanism of wujiadan recipe in the treatment of menopausal syndrome.

Identification and distribution of gonadotropin-releasing hormone-like peptides in the brain of horseshoe crab Tachypleus tridentatus

Gonadotropin-releasing hormone （GnRH） is a crucial peptide for the regulation of reproduction. Using immunological techniques, we investigated the presence of GnRH in horseshoe crab Tachypleus tridentatus. Octopus GnRH-like immunoreactivity, tunicate GnRH-like immunoreactivity, and lamprey GnRH-I-like immunoreactivity were detected in the neurons and fibers of the protocerebrum. However, no mammal GnRH-like immunoreactivity or lamprey GnRH-LII-like immunoreactivity was observed. Our results suggest that a GnRH-like factor, an ancient peptide, existed in the brain of T. tridentatus and may be involved in the reproductive endocrine system.

短期GnRH脉冲治疗对先天性低促性腺激素性性腺功能减退症青少年期男性患者垂体⁃睾丸功能的作用

目的·探讨短期促性腺激素释放激素(gonadotropin-releasing hormone,GnRH)脉冲治疗对先天性低促性腺激素性性腺功能减退症(congenital hypogonadotropic hypogonadism,CHH)青少年期男性患者垂体及睾丸功能的作用。方法·回顾性研究2016年1月—2021年6月接受GnRH脉冲治疗的CHH青少年期男性患者20例,收集患者临床资料。治疗方法为皮下持续脉冲泵给予戈那瑞林治疗1周(20例),其中5例患者持续治疗3个月;剂量为每个脉冲8~10μg,脉冲间隔90 min。于GnRH脉冲治疗前及治疗1周、1个月和3个月时,检测促黄体生成素(luteinizing hormone,LH)、促卵泡刺激素(follicle-stimulating hormone,FSH)、睾酮水平;治疗3个月时测量睾丸体积变化。20例CHH患者均进行了全外显子基因测序。结果·20例CHH患者就诊年龄为14.35(14.08,15.31)岁,临床均表现为幼稚型睾丸、小阴茎,其他还伴有肥胖(12/20)、嗅觉障碍(9/20)、胰岛素抵抗(4/20)、隐睾(4/20)、身材矮小(3/20)等。患者身高为161.79(154.90,173.25)cm,体质量指数为23.80(20.51,27.46)kg/m2,睾丸体积为0.91(0.55,1.25)mL。抑制素B为39.67(11.29,64.97)pg/mL,LH基础值为0.20(0.10,0.30)IU/L,FSH基础值为0.87(0.23,0.89)IU/L,睾酮基础值为0.92(0.38,1.49)nmol/L。持续GnRH脉冲治疗1周后,20例患者LH、FSH的基值和峰值以及睾酮峰值均显著升高(均P<0.05)。其中接受3个月治疗的5例患者治疗1周、1个月、3个月时,LH、FSH的基值和峰值均呈逐渐升高趋势;治疗3个月时LH、FSH基值和峰值,以及睾酮峰值均比治疗前显著增高(均P<0.05),睾丸体积也显著增大(P=0.004)。20例患者中仅14例检测到基因突变,分别为人成纤维细胞生长因子受体1(fibroblast growth factor receptor 1,FGFR1)突变7例、anosmin 1蛋白基因(anosmin 1,ANOS1)突变4例、前动力蛋白2受体(prokineticin receptor 2,PROKR2)突变2例、前动力蛋白2(prokineticin 2,PROK2)突变1例。GnRH脉冲治疗1周对FGFR1突变和ANOS1突变患者垂体-睾丸功能的影响差异无统计学意义。结论·青少年期男性CHH患者接受持续GnRH脉冲治疗1周后垂体-睾丸功能出现应答,治疗3个月有助于该类患者青春期第二性征的诱导。

注射和口服GnRH对小黄鱼人工催产效果的评估

为评估促性腺激素释放激素(Gonadotrophin releasing hormone,GnRH)作为外源激素对小黄鱼(Larimichthys polyactis)的人工催产繁殖效果,以生理盐水注射组为对照,设计促黄体素释放激素(LHRH-A2,2.4μg/kg)注射组、促性腺激素释放激素(GnRH,75μg/kg)注射组、促性腺激素释放激素(6 mg/kg)口服组,通过繁殖生理学等方法比较其产卵情况、卵巢组织生理学变化和血清激素水平综合分析催产效果。结果显示,GnRH注射组在效应时间、产卵持续时间及受精率和孵化率与LHRH-A2注射组无显著差异,但总产卵量和上浮卵重显著升高(P<0.05);GnRH口服组总产卵量和上浮卵重则显著低于LHRH-A2注射组,但受精率和孵化率无显著差异。进一步通过组织生理学分析小黄鱼产卵过程,对照组卵巢发育以Ⅲ期卵巢为主,LHRH-A2注射组、GnRH注射和口服组均可诱导卵巢由Ⅲ期发育至Ⅴ期,促进卵母细胞由Ⅲ和Ⅳ时相发育成熟并排出体外。各实验组经催产激素处理后,血清中促黄体生成素(LH)、卵泡刺激素(FSH)和GnRH迅速上升,至产卵后期仍维持较高水平,各处理组的激素水平变化趋势一致。综上,注射GnRH可有效诱导小黄鱼产卵,且催产效果与注射LHRH-A2无显著差异;口服GnRH也可以诱导小黄鱼产卵,但产卵量和上浮卵重显著低于激素注射组,对其给药方法和剂量需要进一步优化。

A Qualitative Model of the Interaction of Sexual Behavior and Hormone Gene Transcription in Male Blue Gourami during Reproduction

In the present study, a model is suggested to describe hormone control in male blue gourami (Trichogaster trichopterus) along the gonadotropic brain-pituitary- gonad axis (BPG axis) and the hypothalamic-pituitary-somatotropic axis (HPS axis). This model is based on the cloning and transcription of genes encoding hormones of the two axes involved in spermatogenesis during blue gourami reproduction. Gene transcription is affected by environmental, biological, and behavioral factors. Mature males were examined in two different stages—nonreproductive in high-density habitats and reproductive in low-density habitats. Based on gene transcription, gonadotropin-releasing hormone 1 (GnRH1) was involved in controlling spermatogenesis (spermatogonia to spermatids) via the BPG axis in nonreproductive and reproductive stages by controlling follicle-stimulating hormone (FSH), 11-ketotestosterone (11KT) and 17β-estradiol (E2). However, GnRH3 had a larger effect during the reproductive stage via the BPG axis (spermatids to sperm) on luteinizing hormone (LH), 11KT, and 17α-hydroxyprogesterone (17P). At the same time, the HPS axis was involved in spermatogenesis via pituitary adenylate cyclase-activating polypeptide (PACAP) and its related peptide PRP (formerly known as GHRH-like peptide) in the brain, and growth hormone (GH) in the pituitary affected synthesis of insulin-like growth factor 1 (IGF1) in the liver.

GnRH-a联合LNG-IUS治疗子宫腺肌病相关经量过多的随访结局评价

目的:评价促性腺激素释放激素激动剂(GnRH-a)联合左炔诺孕酮宫内缓释系统(LNG-IUS)治疗子宫腺肌病相关经量过多的随访结局。方法:选取西南医科大学附属医院2018年6月至2020年6月接诊的子宫腺肌病相关经量过多患者作为研究对象。研究对象均给予GnRH-a联合LNG-IUS治疗。分析治疗前、治疗后6、12、24、36和48个月的经量图形失血(PBAC)评分、痛经程度视觉模拟(VAS)评分、子宫体积、CA125水平、贫血情况、LNG-IUS带器率。结果:纳入研究患者187例,中位随访时间36.7个月,治疗后48个月累积带器率90.4%,经量过多症状缓解率89.9%。治疗后48个月内,患者的PBAC评分、VAS评分、子宫体积和CA125水平在每个随访时间点与治疗前比较均有显著改善(P<0.01)。结论:GnRH-a联合LNG-IUS可改善子宫腺肌病相关经量过多患者的持续带器率和随访结局。

拮抗剂方案下波塞冬4组患者不同时机添加人重组黄体生成素的临床效果

目的 探讨拮抗剂方案中不同时机添加重组人黄体生成素(rLH)在波塞冬4组患者体外受精/卵胞浆内单精子注射(IVF/ICSI)周期中的临床效果。方法 回顾性分析2017年1月至2022年12月在我中心行拮抗剂方案促排卵并符合波塞冬4组诊断标准的卵巢低反应(POR)患者的临床资料,按照是否添加rLH和添加时机分为4组:对照组(促排卵过程中未添加rLH,n=481)、早卵泡期添加组[在促性腺激素(Gn)启动日添加rLH,n=116]、拮抗剂日添加组(拮抗剂添加日添加rLH,n=100)和晚卵泡期添加组(Gn使用8 d后添加rLH,n=82),比较各组患者的一般情况、促排卵情况、胚胎发育情况和鲜胚移植临床妊娠结局。结果 (1)一般情况:各组间患者年龄、不孕年限、既往IVF/ICSI治疗周期数和不孕类型比较均无显著性差异(P>0.05)。(2)促排卵情况:与对照组比较,早卵泡期添加组Gn刺激天数、Gn总量均显著减少(P<0.05),而晚卵泡期添加组的Gn刺激天数和Gn总量均显著增加(P<0.01),拮抗剂日添加组Gn刺激天数显著减少(P<0.05);拮抗剂日添加组E2水平显著高于对照组(P<0.05),早卵泡期添加组HCG日≥18 mm卵泡数显著高于对照组(P<0.05),早卵泡期添加组和拮抗剂日添加组HCG日内膜厚度均显著高于对照组(P<0.05)。(3)胚胎发育情况:在IVF和ICSI周期中,早卵泡期添加组获卵数、2PN数、2PN卵裂数、D3优胚数和可利用胚胎数均显著高于对照组(P<0.05);在IVF周期中,拮抗剂日添加组获卵数、2PN数和2PN卵裂数均显著高于对照组(P<0.05)。(4)妊娠结局:早卵泡期、拮抗剂日和晚卵泡期添加组鲜胚移植率均显著高于对照组(P<0.05);早卵泡期和拮抗剂日添加组周期取消率显著低于对照组(P<0.05)。结论 对于高龄、卵巢低储备的POR患者,早卵泡期添加rLH可获得更多的卵母细胞、第3天优质胚胎和可利用胚胎,降低周期取消率,获得更多的移植机会。

GnRH激动剂长方案与GnRH拮抗剂方案在不同年龄组、不同反应人群中的新鲜周期临床结局比较

【目的】比较促性腺激素释放激素激动剂(GnRH-a)长方案和促性腺激素释放激素拮抗剂(GnRH-ant)方案在不同年龄组和不同反应人群中行体外受精或单精子卵胞浆内注射-胚胎移植(IVF/ICSI-ET)新鲜周期移植的临床结局。【方法】回顾性分析中山大学附属第三医院生殖医学中心2015年8月28日至2016年12月31日行IVF/ICSI的737个周期,其中Gn RH-a长方案(A组)386个周期,GnRH-ant方案(B组)351个周期。按年龄和获卵数分成a和b两个亚组:a1组(<38岁),a2组(≥38岁);b1组(获卵数≤5个),b2组(获卵数6~15个),b3组(获卵数>15个)。比较患者的临床资料和助孕结局。【结果】(1)A、B两组的受精率、正常受精率、生化妊娠率、流产率均无统计学差异,A组的促性腺激素(Gn)使用天数、Gn总量、人绒毛膜促性腺激素(hCG)日雌二醇(E2)浓度、h CG日内膜厚度、获卵数、成熟卵子数、卵巢过度刺激综合征(OHSS)发生率、胚胎种植率、临床妊娠率均高于B组(P<0.05),B组的新鲜周期移植取消率高于A组(P<0.001)。(2)按年龄分层后,<38岁亚组使用Gn RH-ant方案的胚胎种植率略低于Gn RH-a长方案(32.6%vs 39.8%,P=0.067),两种方案的临床妊娠率没有统计学差异(54.8%vs 50.4%,P=0.429);≥38岁亚组使用Gn RH-ant方案的胚胎种植率低于Gn RH-a长方案(9.7%vs17.9%,P=0.066),使用Gn RH-ant方案的临床妊娠率低于Gn RH-a长方案(19.6%vs 39.1%),差异有统计学意义(P=0.021)。(3)按获卵数分层后,除b1亚组Gn RH-ant方案的胚胎种植率低于Gn RH-a长方案以外(13.1%vs 26.0%,P=0.026),b2、b3亚组使用两种方案的胚胎种植率没有统计学差异;b1、b2、b3三个亚组使用两种方案的临床妊娠率均没有统计学差异,但是b1亚组两种方案的临床妊娠率相差较大(36.6%vs 19.3%,P=0.056)。【结论】总体而言,Gn RH-a长方案的胚胎种植率和临床妊娠率高于Gn RH-ant方案,但Gn RH-ant方案可降低Gn的用量,缩短治疗时间,并有效减少OHSS的发生。对于年轻的卵巢正常反应患者甚至是高反应患者,两种方案有相似的临床结局;而对于高龄等低反应患者,则使用Gn RH-a长方案的胚胎种植率和临床妊娠率更高。

鹅不同繁殖时期GnRH和GnIH基因表达和激素浓度变化分析

旨在研究GnRH和GnIH基因和激素在鹅繁殖过程中的重要作用,本研究以产蛋性能差异显著的四川白鹅和溆浦鹅为试验材料,利用Real-time RCR方法检测产蛋前期、产蛋期和休产期两种鹅的下丘脑、垂体和卵巢组织中GnRH和GnIH mRNA的表达量;利用放射性免疫和酶联免疫吸附测定法测定血清中的GnRH和GnIH激素变化情况。结果表明:GnRH和GnIH基因在四川白鹅和溆浦鹅的下丘脑、垂体和卵巢组织中均有表达;产蛋前期和产蛋高峰期的下丘脑和卵巢组织中,四川白鹅的GnRH mRNA表达量显著(P<0.05)或极显著(P<0.01)高于溆浦鹅,在产蛋高峰期和休产期四川白鹅GnRH激素浓度也极显著(P<0.01)或显著高于(P<0.05)溆浦鹅,在高峰期和休产期四川白鹅GnRH血清浓度分别为51.13和51.10pg·mL-1,溆浦鹅分别为49.52和49.94pg·mL-1;GnIH在两种鹅的变化比较一致,仅在产蛋高峰期,四川白鹅下丘脑组织中GnIH mRNA表达水平极显著低于(P<0.01)溆浦鹅,而两种鹅之间的GnIH激素在各个时期均无显著差异。这提示随着繁殖阶段的改变,在调控两种鹅产蛋性能的作用过程中GnRH可能较GnIH发挥着更为重要的作用,为进一步研究鹅繁殖分子机制奠定基础。

GnRH Ⅱ与GnRH Ⅰ对子宫内膜异位症患者间质细胞分泌VEGF作用的比较(英文)

目的:测定GnRH Ⅱ与GnRH Ⅰ对子宫内膜异位症(EMs)患者离体培养子宫内膜间质细胞分泌血管内皮生长因子(VEGF)的影响,探讨GnRH Ⅱ对EMs患者可能的作用。方法:给予原代培养的EMs患者在位及异位子宫内膜间质细胞不同浓度的GnRH Ⅱ,GnRH Ⅰ类似物(戈舍瑞林,goserelin)处理,同时设对照组(不加GnRH),采用酶联免疫吸附法(ELⅠSA)测定培养液中VEGF浓度,并进行比较。结果:EMs患者离体培养的异位子宫内膜间质细胞经48 h培养,能分泌VEGF,分泌量与在位子宫内膜间质细胞的相近,两者比较差异无统计学意义(P>0.05)。不同浓度的GnRH Ⅱ对EMs患者离体培养的在位和异位子宫内膜间质细胞VEGF的分泌有明显的抑制作用,呈剂量依赖性(P<0.05),且较GnRH Ⅰ类似物(戈舍瑞林)的作用更强(P<0.05)。不同浓度的GnRH Ⅱ对EMs患者离体培养的异位子宫内膜间质细胞VEGF分泌的抑制作用明显强于在位(P<0.05)。结论:EMs患者的异位子宫内膜间质细胞具有分泌VEGF的功能,分泌量与在位子宫内膜的相近,这对EMs的形成和发展可能起重要作用。Gn-RH Ⅱ呈剂量依赖性地抑制异位内膜间质细胞分泌VEGF,其抑制作用明显强于在位,且GnRH Ⅱ明显强于GnRH Ⅰ,为寻找EMs抗血管形成方面的新药治疗提供了新的依据。

GnRH-A主动免疫公兔对垂体GnRHR、FSH-β和LH-β基因表达的影响

目的探讨GnRH-A激动剂(阿拉瑞林)主动免疫对公兔的去势效果和垂体GnRHR、FSH-β和LH-βmRNA表达的影响。方法 30只日本大耳白兔(Oryctolagus cuniculus)随机分为三组(n=10),在实验1组(EG-Ⅰ)和实验2组(EG-Ⅱ)颈部皮下注射1.0mL(100μg/mL)阿拉瑞林抗原乳剂EG-Ⅱ于20d以相同剂量重复注射1次,用荧光定量PCR分析垂体中GnRHR、FSH-β和LH-β mRNA的表达,并测定GnRHR的核苷酸序列。结果 EG-Ⅰ和EG-ⅡGnRH抗体水平高于对照组(P<0.05),EG-Ⅱ在49d达到峰值,显著高于EG-Ⅰ(P<0.05)和对照组(P<0.01),而后开始逐渐下降。28d以后,EG-Ⅱ和EG-Ⅰ血清睾酮浓度低于对照组(P<0.05),且EG-Ⅱ低于EG-Ⅰ(P<0.01);公兔GnRHR的核苷酸为1179bp,同源性达96%。结论阿拉瑞林免疫可以明显提高血清GnRH抗体水平,降低垂体GnRHR、FSH-β和LH-β基因表达,减少睾酮的合成与分泌,从而导致性器官发育受阻,具有明显的作用,加强免疫效果更佳。

鲤鱼sGnRH基因克隆及其在成熟个体的表达分析

采用RACE方法 ,从鲤鱼脑组织克隆了两个差异的sGnRH(salmonGnRH ,[Trp7Leu8]GnRH)cDNAs ,即cDNA1和cDNA2 ,其长度分别为 393和 4 78bp。两个cDNAs都包括一个 2 85bp开放阅读框 ,编码的sGnRH前体为 94个氨基酸残基 ,由一个信号肽、sGnRH十肽和一个由蛋白水解位点 (Gly Lys Arg)连接的促性腺激素释放激素相关肽共 3部分组成。用内含子捕获得到相应的两个差异sGnRH基因 ,即sGnRHgene1和 gene2 ,其基本结构都包括 4个外显子和 3个内含子 ,3个内含子的核苷酸相似性分别为 71 1%、76 1%和 88.0 %。鲤鱼sGnRHcD NAs及基因的基本结构和编码特点与已报道的不同形式GnRHcDNAs和GnRH基因相似 ,由此推测所有类型的GnRH可能来自一个共同的祖分子。Southern杂交进一步证实鲤鱼基因组存在两个不同的sGnRH基因座位。相对定量RT PCR检测发现 ,两个sGnRH基因除在精巢的表达存在差异外 ,在脑区、垂体和成熟卵巢共表达。其中两个sGnRH基因在端脑和下丘脑的表达水平明显高于后脑区。根据sGnRHmRNAs在多个脑区、性腺和垂体的共存推测 ,sGnRH可能对鲤鱼下丘脑 垂体 性腺轴的调节有至关重要作用 。

GnRH-R在大鼠递增负荷训练中的表达及与性激素的关系

:采用动物实验研究法 ,对促性腺素释放激素受体 (GnRH -R)在大鼠递增负荷训练中的表达及与性激素的关系进行分析。结果显示 ,长期大强度负荷使Ta组大鼠垂体GnRH -R表达明显低于C组 ,且同时伴有LH、E2 、P、T明显低于C组 ,FSH也低于C组 ;但是Ta组大鼠下丘脑、血浆 β -EP明显高于C组。休息 1周后GnRH -R未见恢复。认为 ,长期大强度负荷使Ta组大鼠垂体GnRH -R自身调节发生紊乱 ,是引起AMI的重要因素之一 ;过高的 β -EP抑制中枢GnRH分泌 ,是垂体GnRH -R表达减弱的主要抑制神经递质之一 ;推测AMI可能发生在下丘脑水平。

人GnRH及其转运肽基因合成及鉴定

目的人工合成人GnRH及转运肽(TRS)基因。方法根据已发表的人GnRH基因mRNA序列以及转运肽(9个左旋精氨酸)基因核苷酸序列,设计一对核苷酸,再用含7mol/L尿素的变性聚丙烯酰胺凝胶电泳纯化,然后以这对核苷酸3′末端短互补序列退火、补齐,合成长达90bp的目的序列GnRH/TRS。将其亚克隆到pMD18T载体上,构建重组质粒pYC1。酶切鉴定筛选出阳性克隆pYC1,并测序。结果GnRH/TRS序列由90个核苷酸组成,扩增产物与原设计序列同源性达100%。结论已成功合成人GnRH/TRS基因,为进一步研究和应用奠定基础。

拮抗剂方案添加GnRH-α黄体支持治疗后妊娠结局研究

目的:探讨拮抗剂方案添加促性腺激素释放激素激动剂(GnRH-α)行黄体支持对临床结局的影响。方法:回顾分析兰州大学第一医院生殖医学专科医院2017年1月至2017年12月252个拮抗剂方案新鲜周期移植后获临床妊娠的患者的临床资料。根据黄体支持方法不同,将患者分为黄体酮+地屈孕酮组128例,添加GnRH-α组124例(常规黄体酮+地屈孕酮支持的基础上于取卵后第6天加用1次曲普瑞林0.1mg)。比较两组的妊娠率、临床妊娠率、持续妊娠率、早期流产率、异位妊娠率、多胎妊娠率、胚胎种植率、卵巢过度刺激综合征(OHSS)发生率及活产率。结果:两组患者的平均年龄、不孕年限、基础内分泌情况、移植日子宫内膜厚度、移植胚胎数及移植胚胎级别均无统计学差异(P>0.05)。添加GnRH-α组的妊娠率、临床妊娠率及胚胎种植率均高于黄体酮+地屈孕酮组(P<0.05);两组的早期流产率、持续妊娠率、异位妊娠率、多胎妊娠率、OHSS发生率及活产率比较,差异均无统计学意义(P>0.05)。两组妊娠患者的血HCG、E2、PGN比较,差异无统计学意义(P>0.05)。结论:GnRH拮抗剂方案中,添加GnRH-α黄体支持方案可提高临床妊娠率及胚胎种植率,黄体期添加GnRH-α可作为黄体支持的新选择。