重复制动应激对雌性大鼠下丘脑-垂体-卵巢轴的影响

目的: 探索重复制动应激对雌性大鼠下丘脑-垂体-卵巢轴的影响。方法: 40只SD雌鼠随机分为两组(n=20),对照组和实验组,一组正常饲养,一组采取递增负荷束缚应激,每天置于束缚器内制动应激一次(从上午9:00开始),第1日制动2 h,以后采用递增负荷,每日增加0.5 h,持续两周,通过检测体重、脏器系数、动情周期、性激素、病理和相关基因的表达探索其对下丘脑-垂体-卵巢轴的危害。结果: 重复制动应激使雌性大鼠体重下降、动情周期延长,卵巢和子宫的脏器系数和形态发生改变,利用qPCR技术对其相关基因检测,发现下丘脑促性腺激素释放激素、垂体促性腺激素释放激素受体、促卵泡生成素和促黄体生成素mRNA的表达显著下降,卵巢促卵泡生成素和黄体生成素受体 mRNA的表达显著上升,卵巢和子宫雌激素受体mRNA的表达显著下降。结论: 重复制动应激可能通过干扰下丘脑-垂体-卵巢轴的内分泌调节作用,使动情周期紊乱,从而损伤雌性动物的性腺和生殖内分泌功能。     

GnRH及其受体在性成熟前后鲇、黄颡鱼脑、垂体和卵巢中的免疫细胞化学定位

用链霉亲和素 -生物素化过氧化物酶复合物 (StreptAvidinBiotin peroxidaseComplex ,SABC)免疫细胞化学方法 ,使用促性腺激素释放激素 (Gonadotropin releasinghormone ,GnRH)以及促性腺激素释放激素受体 (GnRHR) 2种抗血清对性成熟前后的黄颡鱼 (Pelteobagrusfulvidraco)和鲇鱼 (Silurusasotus)的脑、垂体、卵巢中的免疫活性内分泌细胞进行了免疫细胞化学定位。结果表明GnRH和GnRHR免疫活性在两种鱼的各脑区、垂体、卵巢中均有分布 ;两种鱼在性成熟时它们的下丘脑、垂体和卵巢中的GnRH和GnRHR免疫反应细胞数目和免疫反应强度明显高于性成熟前。本文讨论了GnRH、GnRHR直接或间接参与黄颡鱼和鲇鱼性腺发育成熟调节的可能性及形态学证据。可为下丘脑 垂体 性腺轴、神经 内分泌、GnRH功能的多样性等研究领域提供新的形态学依据。     

营养制剂对间歇性寒冷暴露大鼠子宫、卵巢内HPO轴功能及能量代谢的影响*

目的: 观察一种自行设计的营养制剂对间歇性寒冷暴露SD雌性大鼠下丘脑-垂体-卵巢轴(HPO轴)功能及能量代谢的影响。 方法: 将雌性SD大鼠分为对照组、寒冷暴露组、营养制剂组,每组11只。对照组、寒冷暴露组每日灌胃蒸馏水,营养制剂组每日灌胃营养制剂,灌胃后寒冷暴露组、营养制剂组每日于-10℃冷舱内暴露4 h,持续14 d后,取血清、子宫、卵巢,采用酶联免疫吸附(ELISA)法测定血清卵泡刺激素(FSH)、黄体生成素(LH)等激素指标,采用比色法等检测ATP酶等能量代谢相关指标。 结果: 与对照组相比,寒冷暴露显著上调大鼠卵巢中FSHR、LHR蛋白表达,增强子宫和卵巢Na⁺-K⁺-ATP酶、Ca²⁺-Mg²⁺-ATP酶活性(P<0.05);与寒冷暴露组相比,营养制剂能够下调低氧暴露大鼠卵巢FSHR、LHR蛋白的表达,抑制卵巢和子宫中Na⁺-K⁺-ATP酶、Ca²⁺-Mg²⁺-ATP酶的活性(P<0.05)。 结论: 营养制剂可以有效改善间歇性寒冷暴露对子宫、卵巢内HPO轴相关受体表达和能量代谢的影响。     

促性腺激素释放激素受体的结构及其生物学功能

促性腺激素释放激素受体(GnRHR)属于类视紫红质G-蛋白结合受体(GPCRs)家族的成员.当促性腺激素释放激素(GnRH)与GnRHR结合后,一系列细胞内信号通路被激活从而调节和表达各种生物学功能.本文对GnRHR的基因结构、分子结构、信号调节及生物学功能等进行了综述.     

促黄体素β基因表达中的转导通路及转录因子

促性腺激素释放激素 (GnRH)为下丘脑促垂体激素 ,其脉冲式地释放调节垂体促卵泡素(FSH)和促黄体素 (LH)的合成与释放 ,进而调节动物的生殖活动。LH是由α亚基和 β亚基组成的异二聚体糖蛋白激素 ,其中 β亚基决定激素的特异性。LHβ基因的表达是由GnRH诱发的 ,此过程主要依靠PKC和Ca2 两类信号通路 ,并调节LHβ基因的表达。目前已经发现 ,多种转录因子 ,如早期生长反应基因 (Egr 1)、核受体SF 1基因、Ptx1基因和Sp1基因等 ,通过与LHβ亚基基因的启动子区直接结合 ,而对该基因的表达进行调控。     

促性腺激素抑制激素与生殖轴的关系

下丘脑的促性腺激素释放激素((GnRH)对调控促性腺激素释放十分重要.10年前在鸟类中首先发现了促性腺激素抑制激素(GnIH),它的鉴定提示GnRH不是唯一能直接影响垂体促性腺激素释放的下丘脑神经肤.GnIH及其同系肽广泛存在于鸟类及哺乳动物体内,GnIH在脑部与GnRH神经接触,GnIH可以直接抑制垂体促性腺激素的合成和释放,GnIH及其受体在鸟类和哺乳动物的生殖腺存在.因此GnIH可以在多个水平直接影响生殖轴:脑部、垂体、生殖腺.     

大鼠促性腺激素细胞内蛋白激酶C的改变对黄体生成素mRNA表达的影响

目的:分析大鼠黄体生成素(LH)表达的受体后信号转导机制。方法:促性腺激素(GTH)细胞内蛋白激酶C(PKC)兴奋或抑制后,用促性腺激素释放激素(GnRH)脉冲刺激,然后用实时荧光定量PCR方法测定细胞LH的β亚基(LHβ)mRNA的表达量,并与空白组比较。结果:LHβmRNA随着PKC活性的升高而显著升高,随着PKC活性的降低而显著降低。结论:GnRH脉冲刺激引起LHβmRNA表达,其受体后的信号转导是PKC-Ca2+途径。     

灵长类月经周期的调控

灵长类月经周期的调控与啮齿类不同,在下丘脑没有促性腺激素释放激素(GnRH)的周期性分泌中枢。排卵前促性腺激素(GTH)峰的出现无需下丘脑活动的增强和GnRH分泌的增加。GnRH对垂体GTH的周期性分泌不起控制作用,只起“允许作用”,起控制作用的是卵巢雌激素。雌二醇作用于垂体促性腺细胞的两个功能池,控制GnRH对它们的作用,完成调节GTH分泌的作用。     

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。     

草鱼促性腺激素受体的克隆及表达分析

利用RT-PCR技术从草鱼性腺中克隆出两种促性腺激素受体基因(FSHR和LHR),采用N-J法构建了系统进化树。通过RT-PCR及Real-time PCR技术对FSHR和LHR在成体草鱼不同组织中的时空表达进行分析,发现FSHR和LHR在草鱼卵巢、精巢中表达明显,表达量随着卵母细胞的发育逐渐增高,以上结果为进一步探讨GTHR在鱼类生殖周期中的生理功能奠定了基础。     

Expression of follicle stimulating hormone and luteinizing hormone receptors during follicular growth in the domestic cat ovary

In order to better understand the pituitary regulation of follicular growth in the domestic cat, follicle stimulating hormone (FSH) and luteinizing hormone (LH) receptors (R) were localized and quantified in relation to follicle diameter and atresia using in situ ligand binding on ovarian sections. Expression of FSHR was homogeneous and restricted to follicle granulosa cells from the early antral stage onwards, whereas expression of LHR was heterogeneous on theca cells of all follicles from the early antral stage onward, and homogeneous on granulosa cells of healthy follicles larger than 800 microm in diameter and in corpora lutea. LHR were also widely expressed as heterogeneous aggregates in the ovarian interstitial tissue. Atretic follicles exhibited significantly reduced levels of both FSHR and LHR on granulosa cells, compared with healthy follicles whatever the follicular diameter, whereas levels of LHR on theca cells were lower only for atretic follicles larger than 1,600 microm in diameter. In healthy follicles, levels of FSHR and LHR in all follicular compartments increased significantly with diameter. Although generally comparable to that observed in other mammals, the expression pattern of gonadotropin receptors in the cat ovary is characterized by an early acquisition of LHR on granulosa cells of growing follicles and islets of LH binding sites in the ovarian interstitial tissue.     

Analysis of pituitary gonadotropin concentration in blood serum and immunolocalization and immunoexpression of follicle stimulating hormone and luteinising hormone receptors in ovaries of postmenopausal women

The participation of gonadotropins in ovarian carcinogenesis is well known and is supported by studies with inhibition of pituitary gonadotropin secretion, which results in a diminished risk of cancer. However, there are few data on localization and expression of Follicle Stimulating Hormone and Luteinising Hormone Receptors (FSHR and LHR) in ovaries of healthy postmenopausal women, and their correlation with FSH and LH concentration in blood serum is unknown. The aim of our study was to analyze gonadotropin concentration in blood serum and the expression of FSHR and LHR in ovaries of 207 postmenopausal women. Patients included in the study were divided into three groups depending on the number of years since menopause. We analyzed the concentration of FSH and LH in blood serum and the expression of FSHR and LHR in ovaries. Ovaries of postmenopausal women showed numerous morphological changes in the cortex and medulla when compared to the structure of ovaries of women at reproductive age. In all groups of patients clefts in the surface epithelium and epithelial inclusion cysts were found. The concentration of FSH and LH in the blood serum of women studied increased significantly with time from menopause. Significant differences between analyzed menopausal groups were found. The highest FSH and LH concentration in blood serum were found in women with the longest period of time from menopause. Quantitatively similar expression of FSHR and LHR was found in ovarian surface epithelial cells, in epithelial inclusion cysts and in the connective tissue cells of ovarian stroma. The intensity of the immunohistochemical reaction decreased with time from menopause and with age.     

Chimeric GnRH-LH receptors and LH receptors lacking C-terminus palmitoylation sites do not localize to plasma membrane rafts

Luteinizing hormone and gonadotropin releasing hormone receptors (LHR and GnRHR, respectively) are G protein-coupled receptors with important functions in reproduction. We have developed chimeric GnRHR-LHR that contain the full GnRHR coupled to various forms of the LH receptor C-terminus to explore the role of the LH receptor C-terminus in raft localization of the receptor and signaling. Addition of the full-length LHR C-terminus to GnRHR resulted in localization of the resting chimeric receptor in the bulk membrane rather than plasma membrane rafts as has been reported for the wild-type GnRHR [A. Navratil, S. Bliss, K. Berghorn, J. Haughian, T. Farmerie, J. Graham, C. Clay, M. Roberson, Constitutive localization of the gonadotropin-releasing hormone (GnRH) receptor to low density membrane microdomains is necessary for GnRH signaling to ERK, J. Biol. Chem. 278 (2003) 31593-31602]. With truncation of the LHR C-terminus, approximately 3% of chimeric receptors appeared in low density membrane fractions. Palmitoylation of sites on the LHR C-terminus appears important for raft localization. Mutations to C-terminus palmitoylation sites eliminated translocation of LH receptors from the bulk membrane to rafts upon binding of hCG although these mutant receptors retained the ability to signal via cAMP.     

Luteinizing hormone and follicle-stimulating hormone receptors and their transcribed genes (mRNA) are present in the lower urinary tract of intact male and female dogs

In dogs, one of the side effects of neutering is the development of urinary incontinence. The relationship between neutering and urinary incontinence caused by acquired urethral sphincter mechanism incompetence (USMI) has been reported. Recently, GnRH analogue treatment that suppresses elevated plasma gonadotrophin concentrations post-spaying has been successfully used in incontinent bitches. These data and the fact that non-gonadal tissues may contain receptors for LH (LHR) and FSH (FSHR) suggest that there might be a functional relationship between gonadotrophins and the lower urinary tract in dogs. This study aimed to investigate the presence of LHR and FSHR in the lower urinary tract of intact male and female dogs. Four regions of the lower urinary tract, i.e. (i) body of the bladder, (ii) neck of the bladder, (iii) proximal urethra and (iv) distal urethra were collected from 10 healthy dogs (5 males and 5 anoestrous females). In situ hybridization and immunohistochemistry were performed to characterise the presence of receptor mRNA and receptor protein. Staining was rated semi-quantitatively, incorporating both the distribution and intensity of specific staining. The distribution of receptor expression in different tissue layers (epithelium, subepithelial stroma and muscle) in each region was statistically analyzed. Luteinizing hormone receptor and FSHR mRNA and protein were present in all four regions and in three tissue layers of males and females. Irrespective of region and layer, female dogs expressed significantly higher expression for LHR mRNA (P<0.001), LHR protein (P<0.05) and FSHR protein (P<0.001). The expression of LHR and FSHR mRNA and protein was not uniform and depended on region, tissue layer and gender. The expression of LHR mRNA was higher in the bladder, compared to the urethra (P<0.05). The FSHR mRNA significantly increased from the bladder to the urethra. Protein expression for LHR and FSHR was highest in the proximal urethra (P<0.05). The overall expression for LHR and FSHR at both mRNA and protein levels was highest in the epithelium, intermediate to low in the subepithelial stroma and muscle. A significant interaction between region and tissue layer showed that mRNA and protein expression for LHR and FSHR decreased from the bladder to the urethra in the epithelium and subepithelial stroma. In contrast, it gradually increased from the bladder to the urethra in the muscle. In conclusion, the present study showed that both mRNA and protein for LHR and FSHR were expressed in the canine lower urinary tract, and the expression levels varied between genders and among regions and tissue layers. The presence of these receptors suggests that gonadotrophins have a role in the physiology and/or pathology of the lower urinary tract function in the dog.     

赤腹松鼠血清性激素水平的季节差异

正季节性繁殖是动物适应环境的具体表现,也是动物维持其种群发展的重要策略。生殖激素和动物的繁殖活动关系密切,协调实现繁殖后代的机能(赖平等,2012),如配子的发生、成熟与排出及受精、妊娠、分娩与泌乳等性行为活动。例如雄性甘肃鼢鼠(Myospalax cansus)血清中睾酮(程志兴,2009)和黄山短尾猴(Macaca thibetana)血清中孕酮含量(夏东坡,2007)在交配期间显著升高,     

Expression of luteinizing hormone and follicle-stimulating hormone receptor in the dog prostate

A possible role for gonadotrophins luteinizing hormone (LH) and follicle-stimulating hormone (FSH) in the prostate physiology has been suggested in humans and rats. This study aimed at investigating the presence of receptors for LH and FSH (LHR and FSHR) in the canine prostate. Prostates were collected at post mortem from 6 clinically healthy, sexually intact beagles free from any prostatic disorder. Tissue was sampled from dorsal, middle and ventral regions of each prostate. Immunohistochemical localization was performed on wax-embedded sections using polyclonal antibodies for LHR or FSHR. The pattern and intensity of staining in the parenchyma (glandular epithelium) and stroma were determined using a semiquantitative histologic assessment. Receptors for LH and FSH were consistently present in both the glandular epithelium and the stroma in all tissue samples examined. Expression for both receptors was higher in the glandular epithelium than the stroma of all prostatic regions (P < 0.001). In the glandular epithelium, LHR (P < 0.01) and FSHR (P < 0.05) expression was lower in the lateral than the other regions, and there was no difference between dorsal and ventral regions. However, variations in the expression for LHR and FSHR among prostatic regions were not found in the stroma. These findings have demonstrated that LHR and FSHR are expressed in the dog prostate, and the variation observed in their levels of expression among its regions and tissue layers suggests a potential role of gonadotrophins LH and FSH in the regulation of the prostate physiology, particularly the glandular epithelium.     

卡贝缩宫素联合Bakri球囊压迫对宫缩乏力性产后出血患者卵巢功能和凝血功能的影响

目的:探讨卡贝缩宫素、Bakri球囊压迫联合治疗对宫缩乏力性产后出血患者卵巢功能以及凝血功能的影响。方法:选取2017年2月-2021年3月我院接诊治疗的86例宫缩乏力性产后出血患者,以随机数字表法分为对照组、研究组,分别给予Bakri球囊压迫单一治疗及卡贝缩宫素、Bakri球囊压迫联合治疗。检测对比患者的卵巢功能指标包括LH、FSH、E2水平,记录两组患者舒张压(DBP)、收缩压(SBP)、心率、出血量变化和止血起效时间,测定纤维蛋白原(FIB)、凝血酶原时间(PT)和活化部分凝血活酶时间(APTT),对比两组不良反应发生情况。结果:两组患者DBP、SBP及心率变化波动较小,治疗后数据显示差异无统计学意义(P>0.05);两组患者治疗后2 h、24 h出血量减少,且研究组治疗后2 h、24 h出血量及止血起效时间短于对照组(P<0.05);治疗后3 d,两组患者FIB水平降低,PT、APPT水平升高,且研究组FIB水平低于对照组,PT、APPT水平高于对照组(P<0.05);治疗后3 d,两组患者血清E₂水平升高,FSH、LH水平降低,且研究组血清E₂、FSH、LH水平优于对照组(P<0.05);研究组不良反应发生率(8.89%)高于对照组(4.88%),但是未见显著性差异(P>0.05)。结论:卡贝缩宫素联合Bakri球囊压迫可改善宫缩乏力性产后出血的卵巢功能及凝血功能,提升产后止血效果,增强患者子宫收缩能力。     

Presence of immunoreactive gonadotropin releasing hormone (GnRH) and its receptor (GnRHR) in rat ovary during pregnancy

The present study aims at quantification of gonadotropin releasing hormone (GnRH) by radioimmunoassay, relative expression of its mRNA by real-time PCR accompanied by its cellular localization in the rat ovary by immunonohistochemistry (IHC) during different time points of pregnancy. To determine the involvement of endogenous ovarian GnRH in receptor mediated local autocrine/paracrine functions within the ovary, the cell specific localization of the classical receptor for GnRH (GnRHR) in the ovary by IHC and expression pattern of its mRNA were studied during pregnancy. Receptor expression during each time point within the ovary was reconfirmed by Western blot analysis accompanied by densitometric analysis of the signal intensity. Results reveal that the content of ovarian GnRH reaches its maximum on Day 20. The densitometric analysis of GnRHR receptor expression from Western blot study exhibits a decreasing trend by Day 20. Presence of GnRH and GnRHR mRNA in the ovary indicates the local synthesis of both ligand and receptor in the rat ovary. Differential expression of GnRH/GnRHR in the corpus luteum throughout pregnancy strengthens the hypothesis of the involvement of ovarian GnRH in local ovarian functions by receptor-mediated mechanisms. The expression of GnRH and GnRHR in the atretic antral follicles is indicative of the possible involvement of this decapeptide in processes like follicular atresia. The expression of GnRH/GnRHR in the nonatretic antral follicles and their oocytes requires further in-depth investigation. Collectively, this study for the first time reveals the presence of endogenous ovarian GnRH/GnRHR supporting their possible involvement in local autocrine/paracrine functions during pregnancy.