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。     

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(P0.05),EG-Ⅱ在49d达到峰值,显著高于EG-Ⅰ(P0.05)和对照组(P0.01),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(P0.05),且EG-Ⅱ低于EG-Ⅰ(P0.01);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。     

Alarelin免疫绵羊对垂体GnRHR、FSHR和LHR基因表达与生殖激素分泌的作用

 目的 探讨GnRHago主动免疫对绵羊腺垂体GnRHR、FSHR和LHR表达及生殖激素合成与释放作用,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 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分别皮下注射阿拉瑞林抗原200μg、300μg和400μg,0d和14d各1次,共2次;EG-Ⅳ和EG-Ⅴ分别皮下注射阿拉瑞林抗原200μg、300μg,0d、7d、14d和21d各1次,共4次;对照组皮下注射药物的溶媒,0d和14d各1次,共2次。于70d在颈动脉放血处死绵羊,无菌切取腺垂体、子宫及卵巢。用ELISA测定血清GnRH抗体浓度,以激素检测试剂盒分别测定血清GnRH、FSH、LH和E2浓度。提取腺垂体总RNA,PCR扩增与反转录,实时荧光定量PCR（FQ-PCR）,分析GnRHR、FSHR和LHR mRNA的表达。 结果 绵羊腺垂体中有丰富的RNA。特异性条带扩分别为2150 bp,1100bp和180bp左右。实验组GnRHR、FSHR和LHR mRNA值均低于对照组,EG-Ⅳ和EG-Ⅴ的mRNA值低于EG-Ⅰ和EG-Ⅱ。实验组血清GnRH浓度逐渐降低值谷值,之后上升,70d 时达到免疫注射前水平。实验组血清FSH浓度始终高于对照组（P＜0.05）。35d时EG-Ⅳ和EG-Ⅴ低于EG-Ⅰ、EG-Ⅱ和EG-Ⅲ。实验组和对照组血清E2含量无显著差异。结论 阿拉瑞林主动免疫可抑制垂体GnRHR mRNA、FSHR mRNA和LHR mRNA的表达,促进垂体合成与释放FSH,增加血清FSH含量,降低GnRH和LH浓度,对E2浓度无明显影响。增加阿拉瑞林抗原的免疫剂量和注射次数,作用更加明显。     

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。     

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。     

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。     

促性腺激素释放激素激动剂改善子宫内膜异位症患者生育力的作用机制及应用

促性腺激素释放激素激动剂（GnRHa）为治疗子宫内膜异位症（EM）的常用药物之一。EM可多方面对女性生育力产生不利影响,而GnRHa可通过改善EM患者腹腔及卵泡微环境、提高子宫内膜容受性及改善输卵管功能等作用提高EM相关不孕患者生育力,并在联合腹腔镜手术及辅助生殖技术（ART）治疗改善EM不孕患者妊娠结局中具有一定价值。     

促性腺激素释放激素激动剂治疗子宫内膜异位症的进展

促性腺激素释放激素激动剂（gonadotropin releasing hormone agonists,GnRH-a）有效的治疗子宫内膜异位症已得到普遍的认可及应用。随着对GnRH-a的作用机制及临床应用方面的不断认识,GnRH-a治疗子宫内膜异位症有了新的进展,随之产生的反加疗法及时减轻了GnRH-a治疗子宫内膜异位症的不良反应,即血管运动综合症及骨质疏松等。     

脑室注射L-谷氨酸对成年大鼠下丘脑GnRH含量的影响

目的 :观察脑室注射L -谷氨酸 (L -Glu)对成年雄性Wistar大鼠下丘脑促性腺激素释放激素(GnRH)含量的影响。方法 :摘取下丘脑组织 ,匀浆化 ,用RIA法检测匀浆上清液中GnRH的含量。结果 :脑室分别注射 0 0 1176、0 1176、1 176 0 μg/ 2 0 μl-1 -1L -Glu后 40分钟 ,下丘脑GnRH含量依次为 1 5 9± 0 41、0 88± 0 34、0 70± 0 42ng/ 10mg湿重 ,均显著低于盐水对照组 (P <0 .0 1) ;脑室注射 0 1176 μg/ 2 0 μl-1·-1L -Glu后 2 0、40、12 0分钟 ,下丘脑GnRH含量依次为 0 99± 0 37、0 88± 0 34、1 2 6± 0 39ng/ 10mg湿重 ,亦均显著低于盐水对照组 (P <0 .0 1)。脑室注射L -Glu对下丘脑GnRH含量的降低作用呈现剂量与时间依从关系。而脑室注射3H -Glu 2 μCi/ 2 0 μl-1·-1后 40分钟发现 ,大脑、小脑、垂体、下丘脑内侧基底部 (MBH)和视前区 (POA) 5个不同部位脑组织中以MBH对3H -Glu的摄取量最大 (10 6 9 82± 490 33cpm/ 10mg湿重 )。结论 :L -Glu可能参与了大鼠下丘脑GnRH神经元功能活动的调节。     

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.     

Comparison of the GnRH agonist and antagonist protocol on the same patients in assisted reproduction during controlled ovarian stimulation cycles

Despite the fact that both gonadotropin-releasing hormone (GnRH) agonist and antagonist protocol are effective in suppressing the incidence of premature luteinizing hormone (LH) surges through reversibly blocking the secretion of pituitary gonadotropins, the exact impact of these two distinctive protocols on the clinical setting of patients for in vitro fertilization and embryo transfer (IVF-ET) treatment, however, remained controversial. We thus in the present report conducted a retrospective study to compare the impact of GnRH agonist and antagonist protocol on the same patients during controlled ovarian stimulation cycles. A total of 81 patients undergoing 105 agonist and 88 antagonist protocol were analyzed. We failed to detect a significant difference between two protocols for the difference in duration of ovarian stimulation, number of recombinant FSH (Gonal-F) ampoules used, number of oocytes retrieved, serum levels for estradiol (E2) and progestone (P), thickness of endometrium, and the zygote- and blastocyst-development rate. It is seemly that high quality embryo rate was higher in the antagonist protocol, but the data did not reach a statistical significance. Nevertheless, Implantation rate and clinical pregnancy rate were significantly higher in the antagonist protocol (10.64% and 30.26%, respectively) than that of the agonist protocol (5.26% and 15.82%, respectively). Our data also suggest that the GnRH antagonist protocol is likely to have the advantage for improving the outcome of pregnancy in those patients with a history of multiple failures for the IVF-ET treatment.     

Comparative stimulatory effect of gonadotrophin releasing hormone (GnRH) and GnRH agonist upon pulsatile human chorionic gonadotrophin secretion in superfused placental explants: reversible inhibition by a GnRH antagonist.

The roles of gonadotrophin releasing hormone (GnRH) and a GnRH agonist (GnRHa) (D-Ala6-Met-Leu7-Pro-N-ethyl-amide) in controlling pulsatile human chorionic gonadotrophin (HCG) secretion by superfused placental explants in the first trimester were examined. One minute pulses of both GnRH and GnRHa had a biphasic effect upon pulsatile HCG secretion. GnRHa was maximally effective at 10(-10) M concentration, at 10(-11) M the effect was mild while at 10(-8) M, no effect was noted. GnRH exerted a maximal stimulatory effect at 10(-8) M; at 10(-10) M no effect was seen, while at 10(-7) M the effect was mildly stimulatory. This was evaluated by carrying out both a between and within channel type of analysis. The effect of a GnRH antagonist GnRH(ant) upon GnRH and GnRHa-induced HCG secretion was examined. Explants were incubated overnight with 10(-8) M GnRH(ant), which was also continuously administered during superfusion. The addition of 1-min pulses of GnRH and GnRHa during the exposure to GnRH(ant) failed to stimulate pulsatile HCG secretion. This effect was reversible since the response to GnRH was restored within 10 min after stopping GnRH(ant) administration. In addition, by the third cycle, co-administration of GnRH(ant) for 2 min together with 10(-10) M GnRHa for 1 min completely blocked the GnRHa-induced effect. Continuous administration of 10(-8) M GnRH(ant) decreased spontaneous HCG pulse amplitude and the area under the curve but failed to modify pulse frequency. In conclusion, GnRH appears to exert a receptor-dependent stimulatory effect upon pulsatile HCG secretion in superfusion in the first trimester placenta.(ABSTRACT TRUNCATED AT 250 WORDS)     

Age- and sex-specific changes in naloxone-induced luteinizing hormone secretion and Fos expression in gonadotropin-releasing hormone neurons of gonadectomized rats

In the present study, we examined sex-specific changes in luteinizing hormone (LH) secretion and Fos expression in gonadotropin-releasing hormone (GnRH) neurons in response to naloxone in young (3 months old) and old (24 months old), gonadectomized male and female rats. We revealed by immunocytochemistry that, regardless of age and sex, naloxone significantly increased the number of GnRH neurons expressing Fos, which was associated with increased LH secretion. Additionally, although the magnitude of the increase in Fos-expressing GnRH neurons did not change in old males compared to young males, it was attenuated by almost half in old females compared to young females. LH levels decreased 60% in old males compared to young males and 15% in old females compared to young females. These results suggest LH secretion is impaired with age, but the ability of GnRH neurons to be stimulated by naloxone is preserved. However, the opioid-controlling mechanism is more fragile in females than males during aging.     

Half-dose,long-acting gonadotropin-releasing hormone agonist (Diphereline) is comparable with daily injections of short-acting gonadotropin-releasing hormone agonist (Suprefact) in IVF/ICSI cycles

Introduction

The aim of the study was to compare the efficacy of half-dose, long-acting GnRH analogue (Diphereline) with Suprefact in IVF/ICSI (in vitro fertilization/intracytoplasmic sperm injection) cycles.

Material and methods

In this randomized clinical trial performed in Royan Institute, 126 infertile women who were first time candidates for IVF/ICSI were enrolled. Patients were randomly divided into two groups by using a random number table. In one group, 62 patients received a single half dose, 1.87 mg Diphereline, in mid-luteal phase. In the other group, 64 cases were treated with buserelin from the previous mid-luteal phase. P value less than 0.05 was considered significant.

Results

The mean age of patients in the Diphereline and Suprefact groups was 27.9 ±3.6 and 29.6 ±3.5 years, respectively (p = 0.01). In the Diphereline group, the mean number of used gonadotropins was 25.6 ±12.1 ampoules, while in the second group it was 25.9 ±8.5 ampoules. Numbers of retrieved and MII oocytes were significantly higher in the Diphereline group (12.1 ±6.3 and 9.6 ±5.5) in comparison to the Suprefact group (9.4 ±6.4 and 7.2 ±5.1). Although the number of developed embryos in the Diphereline group was statistically higher than in the Suprefact group (6.1 ±3.9 vs. 4.7 ±3.4, p = 0.04) there was no significant difference in pregnancy rate (37.1%, 95% CI [26.16-49.54] vs. 37.5%, 95% CI [26.67-49.75]).

Conclusions

A half-dose, long-acting GnRH agonist can be successfully used in ovarian stimulation and produces a higher number of MII oocytes and embryos. The pregnancy rates with this method are acceptable.     

重组GnRH主动免疫对公猪体重及生长激素和IGF-I的影响

目的:探讨重组GnRH六聚体-麦芽糖结合蛋白(MBP-GnRH-6)主动免疫对公猪体重、生长激素和胰岛素样生长因子-I的影响。方法:7头公猪在9周龄时用重组GnRH主动免疫,17周龄时加强免疫,免疫和屠宰前1天称猪体重,酶联免疫分析法检测血清猪生长激素,放射免疫分析法检测血清IGF-I和睾酮水平,Pearson相关分析血清睾酮与IGF-I的相关性。结果:MBP-GnRH-6主动免疫猪血清睾酮浓度显著地低于阳性对照组(P<0.05),体重显著重于阴性对照组(P<0.05),但不影响公猪血清生长激素水平(P>0.05)。在17~21周龄时,免疫组公猪血清IGF-I浓度显著低于阳性对照组(P<0.05),21~25周龄时免疫组公猪血清IGF-I显著高于阴性对照组(P<0.05)。相关分析表明血清睾酮与IGF-I呈正相关。结论:MBP-GnRH-6主动免疫改变了公猪生长性能,增加体重,其作用途径可能是睾酮通过调控血清IGF-I水平来调控的。     

促性腺激素释放激素类似物对乳腺癌细胞株化疗敏感性的影响

目的： 探讨促性腺激素释放激素类似物（GnRHa）对乳腺癌细胞株（MCF-7和MDA-MB-231）化疗敏感性的影响。方法：不同浓度的GnRHa（曲普瑞林,triptorelin）（10^－9 mol/L、10^－8 mol/L、10^－7 mol/L、10^－6 mol/L、10－5 mol/L）分别作用于MCF-7和MDA-MB-231细胞24 h、96 h和168 h后，用CCK-8方法检测细胞活性。用或不用GnRHa（10^－5 mol/L）处理96 h后，分别加入5-氟尿嘧啶（5-FU）或表阿霉素(EPI)作用24 h，用CCK-8法检测细胞抑制率。用RT-PCR检测GnRHa（10^－5 mol/L）作用168 h后GnRH受体、PCNA和MDR1 mRNA表达水平。结果：不同浓度GnRHa作用不同的时间后对乳腺癌细胞活性无影响。GnRHa（10^－5 mol/L）作用96 h后，5－FU和EPI对两种细胞的IC50不改变；GnRHa（10^－5 mol/L）不影响5－FU（MCF-7细胞0.5 g/L，MDA－MB－231细胞0.5 g/L）和EPI（MCF-7细胞1.2 mg/L,MDA－MB－231细胞0.8 mg/L）对两种细胞的抑制作用（P>0.05）。GnRHa（10^－5 mol/L）作用168 h后，MCF-7细胞的PCNA mRNA表达无改变。而在MDA-MB-231细胞，PCNA表达升高，差别有统计学意义（P<0.05）。在MCF-7对照组中，MDR1 mRNA有弱表达。GnRHa作用后，抑制了MDR1 mRNA表达。MDA-MB-231细胞GnRHa作用前后， MDR1 mRNA均无表达。结论：GnRHa不影响乳腺癌细胞株对5-FU和EPI的敏感性。GnRHa可能通过下调MDR1 mRNA表达水平，减弱MCF-7细胞的耐药性。     

Endocrinology: The effect of pre-treatment with a gonadotrophin-releasing hormone agonist on reproductive functions in mature cycling rats

In order to investigate the performance of follicles in a ratmodel in which gonadotrophin-releasing hormone agonist (GnRHa)was used for hypothalamic—pituitary—ovarian axissuppression, three groups of mature cycling rats were studied.One group was treated with buserelin followed by pregnant mare'sserum gonadotrophin (PMSG), and the second group was treatedwith PMSG alone. Both these hormonally treated groups receivedhuman chorionic gonadotrophin for induction of ovulation. Thethird group received no hormonal treatment. The average numberof ovulated oocytes recovered from rat oviducts pre-treatedwith GnRHa was significantly higher than that in rats treatedwith the gonadotrophin alone, in spite of the larger numberof pre-ovulatory follicles present in the gonadotrophin-treatedgroup. The morphology of both the pre-ovulatory and the post-ovulatorycumulus—oocyte complexes in the three groups appearedsimilar. No difference in the capacity of follicles of the threegroups to synthesize progesterone in vitro in response to luteinizinghormone could be observed. We conclude that ovarian morphologyand function are not impaired by pre-treatment with buserelin.