Microscopic characterization of follicular structures in letrozole-induced polycystic ovarian syndrome in the rat

BACKGROUND: Our objective was to characterize the tissular distribution of relevant cytoskeletal proteins, cellular adhesion molecules and proliferation markers and conduct a histomorphometrical study of the follicular wall of letrozole-induced polycystic ovaries. METHODS: Twenty rats were divided into two groups: a control group (C) of ten rats that received vehicle only (0.9% NaCl solution) once daily p.o. and a treatment group (T) of ten animals administered letrozole at a concentration of 1 mg/kg p.o. dissolved in 0.9% NaCl solution once daily during 21 days. Twenty four h after the last administration, all animals were sacrificed. Control animals were sacrificed in proestrous (n = 5) and diestrous (n = 5). Serum hormone levels, histomorphometrical changes and immunoexpression of intermediate filaments (vimentin, cytokeratins and desmin), cadherins and proliferation cellular nuclear antigen were examined. RESULTS: The granulosa cell layer of cystic follicles had a greater significant immunostaining for vimentin and cytokeratins. Immunohistochemical localization of desmin was restricted to the theca externa. Positive immunoreactivity for cadherins rises gradually and significantly, together with the follicular development, and immunoreactivity was comparatively stronger in follicular cysts. A significantly higher immunostaining for PCNA cells was observed in secondary and tertiary follicles as compared with atretic and cystic follicles. An increase in the LH, FSH and testosterone serum concentrations was observed in letrozole-treated rats. Estradiol and progesterone showed a considerable reduction. CONCLUSIONS: The changes observed are probably due to structural and functional alterations that occur during the process of cystogenesis and may be associated with important modifications in the expression of cytoskeletal proteins, cellular adhesion molecules and proliferation markers that may be essential for proper cellular functioning.     

大鼠胚胎卵巢异体异位移植的探讨

为了研究胚胎卵巢移植后的存活和生长发育情况 ,以及不同移植部位对胚胎卵巢的影响 ,将大鼠胚胎卵巢分别移入去势雌鼠的颈部皮下及肾被膜下 ,HE观察颈部移植物移植后 5d处于坏死状态 ,含有大量红细胞 ;1 0d周围有结缔组织包裹 ,内有原始卵泡 ;2 5d及以后移植物内存在大量发育卵泡并可见间质腺及黄体。肾被膜移植物移植后 5d呈结缔组织样 ;1 0d出现大量原始卵泡结构 ;2 5d及以后可见发育卵泡、间质腺和黄体。细胞化学显示发育卵泡的 3 β羟甾脱氢酶呈阳性反应。移植后 2 5d左右阴道涂片出现动情周期。结果说明胚胎卵巢能在异体异位存活并可发育成具有内分泌功能的腺体。     

PCOS大鼠卵巢中 kisspeptin／kiss1r 系统的表达变化及对卵泡发育的影响

目的：对多囊卵巢综合征（PCOS）大鼠卵巢中kisspeptin／kiss1r系统的表达变化及对卵泡发育的影响进行研究。方法采用腹腔注射来曲唑构建PCOS大鼠模型,酶联免疫吸附（ELISA）法测定血清中性激素水平,苏木精‐伊红（H‐E）染色检测卵巢的形态学改变,免疫组化检测卵巢中kisspeptin／kiss1r、卵泡刺激素受体（FSHR）的蛋白水平变化。结果从灌胃后第10天开始,PCOS组大鼠体重明显增加,血清中睾酮（T）、黄体生成素（LH）的水平明显升高,卵泡刺激素（FSH）水平无明显改变;大鼠卵巢皮质中含有较多的小卵泡及闭锁卵泡,多数卵泡成囊性扩张,颗粒细胞层数明显减少;卵巢中kisspeptin／kiss1r、FSHR均在细胞质表达,kisspeptin／kiss1r主要在颗粒细胞、膜细胞和黄体中表达,而FSHR仅表达在颗粒细胞。PCOS组卵巢中kisspeptin／kiss1r、FSHR表达明显减少。结论在PCOS卵泡发育中,卵巢表达的kisspeptin／kiss1r系统发挥重要的作用,为PCOS的临床治疗和基础研究提供新思路。     

Histological study on norethynodrel treated rat ovary

In an attempt to disclose the antiovulatory mechanism of norethynodrel, a progestational compound having slight estrogenic action, a histological study of treated rat ovaries was made. 4 test animals were injected daily for 14 days with 1 mg of norethynodrel in .3 ml sesame oil. All were killed 24 hr after the last injection. Ovaries of treated rats showed follicular degeneration (cystic atresia), interstitial cell degeneration, and cellular hypertrophy of lutein cells. Immature follicles did not show remarkable changes. Findings suggest that norethynodrel may cause hypersecretion of luteotrophic hormone (LTH) and hyposecretion of follicle-stimulating hormone (FSH) and luteinizing hormone (LH) from the hypophysis. Also noted was that secretion of 20-alpha-OH-P decreased greatly with norethynodrel, but the progesterone level showed no change, suggesting that ovarian interstitial cells may synthesize 20-alpha-OH-P from sterol precursors not involving progesterone. The antiovulatory mechanism may depend on direct or indirect interference with the biosynthesis of 20-alpha-OH-P.     

PLASMA PROGESTERONE LEVELS IN NORMAL AND PREGNANT CHINESE WOMEN AND EFFECTS OF CONTRACE.PTIVES ON THEM

Measurement of progesterone levels in peri- pheral plasma during normal human menstrual cycles and pregnancy by competitive protein binding assay shows that during the follicular phase mean plasma progesterone concentrations are 0.39 + 0.14 ng/ml. Its maximal value during the luteal phase is 8.9+ 1.87 ng/ml. On the day before or on the first day of menstruation the level falls t0 1.79+1.21 ng/ml. In early pregnancy the progesterone concentrations are similar to those during the luteal phase. There- after it rises gradually and peaks during late pregnancy, the maximal value being over 130 ng/ml. Anordrin can decrease plasma progesterone levels and shorten the luteal periods of treated women. AF-75 shows no ovulation or development conspicuous effect on of the corpus luteum.     

葛根素对多囊卵巢综合征大鼠卵巢转化生长因子-β1表达的影响

目的:探讨葛根素对多囊卵巢综合征(PCOS)大鼠卵巢组织中转化生长因子-β1(TGF-β1)表达的影响。方法:40只SD雌性大鼠采用硫酸普拉睾酮钠皮下注射建立PCOS模型成功后随机分为模型对照组,葛根素高、低剂量组[120 mg/(kg.d)、80 mg/(kg.d)],另取20只健康雌性大鼠作为空白对照组。放射免疫法测定血清雄激素(Testosterone)水平变化观察卵巢组织形态学改变及排卵情况,免疫组化法测定卵巢TGF-β1表达水平。结果:与模型对照组相比,葛根素高、低剂量组血清雄激素水平和卵巢TGF-β1表达较低,但组间比较无显著差异(P均<0.05);2组卵泡膜及间质细胞增生明显改善,卵泡颗粒细胞层数明显增多,且囊性扩张卵泡及闭锁卵泡减少,有优势卵泡和黄体形成。结论:葛根素可能通过下调卵巢局部TGF-β1表达水平从而降低PCOS大鼠雄激素水平,减轻卵巢多囊样改变。     

Localization of cystathionine β synthase in mice ovaries and its expression profile during follicular development

Background In vitro fertilization (IVF) researches have suggested that cystathionine β synthase (CBS) is involved in oocyte development. However, little is known about the regional and cellular expression patterns of CBS in the ovary. The purpose of this study was to analyze the localization of CBS in mice ovaries and to investigate the expression profile during follicular development. Methods We used in situ hybridization and immunohistochemical analysis to determine CBS expression in the ovaries of female Balb/c mice. Then the follicles were collected from F1 (C57BL×Balb/c) mice and cultured in vitro. With the method of semi-quantitative RT-PCR, we also investigated the expression profile of CBS during follicular development. Results CBS was absent in the oocytes, although it was ubiquitously expressed in the ovary with the strongest expression in follicular cells at all stages. In late antral follicles, CBS expression was markedly higher in granulosa cells located close to the antrum and in cumulus cells around the oocyte. The semi-quantitative RT-PCR showed that CBS mRNA was detected in follicles at all stages in vitro. In cumulus-oocyte complexes superovulated, CBS expression also increased rapidly. Conclusions CBS was located mainly in the follicular cells in the ovaries. The level of CBS expression is high in follicles during folliculogenesis in mice. Differences in the CBS expression profile between oocyte and follicular cells suggest a role for CBS as a mediator in interactions between oocyte and granulosa cells.     

Effect of estrogen deprivation on follicle/oocyte maturation and embryo development in mice

Estrogenissynthesizedbygranulosacellsunderthecontroloffolliclestimulatinghormone (FSH )andluteinizinghormone (LH) Itisbelievedthatestrogenplayspivotalrolesintheregulationoffollicle/oocytematurationandoocytefertilizability Estrogenisalsoinvolvedinthefunctionalpreparationofthefallopiantubesforsubsequentgameteinteraction ,inearlyembryonicdevelopmentoccurringinthetubalmicroenvironment,andinthepreparationoftheuterusforimplantation However,areevaluationoftherolesofLH ,FSH ,andestrogenhasbeensug…     

低氧预处理人脐带间充质干细胞治疗早发性卵巢功能不全小鼠的效果研究

目的 探索人脐带间充质干细胞（human umbilical cord mesenchymal stem cells,hUCMSCs）及其经低氧预处理后对早发性卵巢功能不全（premature ovarian insufficiency,POI）小鼠的治疗作用。方法 从脐带中分离、培养出hUCMSCs,利用白消安和环磷酰胺诱导形成小鼠POI模型,分别使用正常hUCMSCs和经低氧预处理的hUCMSCs对POI小鼠进行治疗。实验结束时,测量各组（n=15）小鼠血浆中雌二醇（estradiol,E2）、卵泡刺激素（follicle stimulating hormone,FSH）及抗苗勒管激素（anti Mullerian hormone,AMH）浓度,对卵巢进行组织病理学检查,计数卵泡数量。结果 经过hUCMSCs和低氧预处理hUCMSCs治疗的小鼠血浆E2及AMH浓度,与模型对照组比较,呈升高趋势;且AMH浓度差异有统计学意义（P<0.05）;与模型对照组比较,hUCMSCs和低氧预处理hUCMSCs治疗组小鼠卵巢组织中生长卵泡明显增多、闭锁卵泡明显减少（P<0.05）;卵巢颜色红润,尺寸增大,几乎没有坏死点,卵泡大、透明度好,整体来看与正常对照组卵巢状态相似;且经低氧预处理hUCMSCs治疗的小鼠,其生长卵泡和闭锁卵泡数量更趋于正常。结论 hUCMSCs移植及hUCMSCs低氧预处理后移植治疗均能够有效恢复POI小鼠的激素浓度,促进卵巢功能的恢复。     

逍遥散对慢性应激肝郁证模型小鼠卵泡发育障碍的影响

目的 观察逍遥散对慢性应激致肝郁证模型小鼠卵泡发育障碍的作用。 方法 将雌性ICR小鼠随机分为正常组、模型组和逍遥散组,每组20只。采用慢性不可预见应激复制肝郁证小鼠模型,促性腺激素促排方法观察排卵数,苏木精-伊红染色法观察各级卵泡发育情况,酶联免疫吸附试验检测卵泡内分泌雌二醇、孕酮功能。 结果 与正常组比较,模型组小鼠排卵数、正常次级卵泡和窦卵泡数、血浆雌二醇和血清孕酮水平显著减少（P<0.05）,闭锁次级卵泡和窦卵泡数显著增加（P<0.05）;逍遥散组上述指标较模型组显著逆转（P<0.05）。 结论 逍遥散可改善慢性应激致肝郁证小鼠的卵泡发育,促进小鼠排卵和生殖内分泌功能。     

武汉地区正常育龄妇女月经周期血清中七种激素的放射免疫测定

选抒12例正常月经周期的武汉地区妇女,研究她们的内分泌图象,采用放射免疫测定法测定每天血清中促黄体生成激素(LH)、促卵泡生成素(FSH)、垂体催乳素(PRL)、雌二醇(E_2)、孕酮(P)、睾丸酮(T)和皮质激素(C)的浓度。其中LH、FSH、E_2和P的周期变化和水平与其他文献报告的结果相同,月经中期LH峰值是50mIU/ml,FSH峰值为12mIU/ml,排卵前E_2峰值为252pg/ml,P值为16ng/ml,在大多数周期中,PRL在月经中期水平是高的,但黄体期PRL平均水平高于滤泡期,月经中期T和C值无明显变化,而滤泡期C水平(91ng/ml)高于黄体期(81ng/ml)。     

小鼠双侧卵巢自体移植的组织化学研究

对小鼠双侧自体移植的卵巢进行组织化学研究，探讨其生长规律。结果表明：移植第２～４ｄ，卵巢的原有组织处于退化状态，边缘偶见窦前卵泡，仅散在分布酶染色阳性物质。移植第７～１４ｄ，卵巢中出现窦状卵泡，酶染色较强阳性，大多数动物出现动情周期。移植第１４ｄ以后，卵巢内含发育不同阶段的卵泡、黄体和大量间质腺，酶分布广泛，染色强阳性。研究结果提示双侧自体移植卵巢能够生长发育，并可能分泌性激素。     

柴胡加龙骨牡蛎汤对大鼠心理应激反应的影响

[目的]观察柴胡加龙骨牡蛎汤对急性心理应激大鼠旷场行为和血浆促肾上腺皮质激素的影响。[方法]采用大鼠急性心理应激动物模型,在应激前用两种不同浓度的柴胡加龙骨牡蛎汤胃饲7d,观察心理应激后血浆促肾上腺皮质激素含量的动态变化以及大鼠旷场行为的改变,并与对照组相比较。[结果]与正常对照组比较,应激大鼠在旷场中的穿行格数和直立次数明显增加,血浆促肾上腺皮质激素浓度显著升高,而两种不同浓度的给药,对此反应均有抑制作用。[结论]柴胡加龙骨牡蛎汤对急性心理应激大鼠的心理行为反应和生理反应具有调节作用。     

α-硫辛酸对SD 大鼠下丘脑-垂体-肾上腺皮质轴形态和功能的影响

目的 研究α-硫辛酸对正常和糖尿病大鼠下丘脑-垂体-肾上腺皮质功能和超微结构的影响.方法 应用放射免疫法分别观察了低、中、高剂量(1、20、100 mg/kg)α-硫辛酸(100 mg/kg)连续腹腔注射3周,对SD大鼠血浆CRH、ACTH、皮质酮水平的影响.并运用电镜观察用药前后大鼠垂体、肾上腺皮质超微结构的变化.结果 正常和糖尿病大鼠低中高剂量α-硫辛酸处理组CRH水平与相应对照组相比显著降低;正常大鼠低中高剂量硫辛酸处理组ACTH水平比相应对照组明显降低(P<0.05);正常大鼠低中高剂量α-硫辛酸处理组和糖尿病大鼠中高剂量α-硫辛酸处理组皮质酮水平比相应对照组显著降低(P<0.05).α-硫辛酸处理对正常大鼠下丘脑-垂体-肾上腺皮质超微结构无明显影响.糖尿病大鼠垂体细胞和肾上腺皮质细胞出现功能受抑制状态,α-硫辛酸处理21d后垂体促肾上腺皮质细胞和肾上腺皮质细胞代谢较处理前明显活跃.结论 α-硫辛酸能在不同层次直接或间接地发挥对正常和糖尿病大鼠HPA轴的抑制作用.     

免疫应答对血浆ACTH和皮质酮水平的影响

ＮＤＶ腹腔免疫小鼠后３ｈ血浆ＡＣＴＨ水平升至高峰，６ｈ下降，１２ｈ基本恢复至正常水平，ＮＤＶ免疫后血浆皮南酮（ＣＳ）６ｈ升至高峰，１２ｈ已有大幅度下降，２４ｈ后基本恢复至正常水平，ＮＤＶ可使大鼠血浆ＡＣＴＨ和ＣＳＣ水平增加，白喉类 足垫皮下免疫后小鼠血浆ＣＳ水平有类似变化。白喉类毒素和绵羊红细胞（ＳＲＢＣ）免疫后７天均可升高小鼠血浆ＣＳ水平；ＮＤＶ免疫后７天则可降低小鼠血浆ＣＳ水平。     

己烯雌酚促验进实动物卵泡发育同步化及其机制的初步研究

目的观察人工合成的非甾体化合物己烯雌酚（DEs）对性成熟前小鼠和大鼠卵巢中卵泡发育的影响,以及出现有卵泡同步化机理。方法取性成熟前雌性小鼠和大鼠,腹腔注射一定剂量的DES,然后取卵巢经连续切片和HE染色后,统计整个卵巢内三级卵泡数量。同时,取卵巢利用免疫组织化学和免疫印迹方法,分别检测泛素末端水解酶LI（UCH-L1）,c-Jun结合蛋白l（Jabl）和细胞周期素依赖激酶抑制因子p27KipI在小鼠卵巢中的定位和含量。结果UCH-LI定位在卵巢中各级卵泡的卵母细胞中,Jab1分布于次级和三级卵泡的卵母细胞核中,而p27KipI广泛分布于卵巢各类细胞中。免疫组化结果显示,仅就卵母细胞而言,由于DES的处理,UCH—L1和Jab1的表达信号增强,而p27KipZ的信号变弱,与肿瘤细胞中的情况类似。然而,免疫印迹分析结果表明,以上3种蛋白在卵巢中相对b-actin的含量因DES处理而均呈现一定程度的上升变化趋势。青春前期的小鼠,在1-4周龄期间,卵巢中UCH．L1和p27KiPI的蛋白量随着周龄增加的趋势,而Jab1的变化趋势不明显。结论腹腔注射DES或导致卵巢内卵泡发育同步化,其中涉及的机理可能是,UCH-L1及其关联蛋白Jab1导致p27KiPI在卵母细胞核中的含量下降,进而导致了多个卵泡的卵母细胞同时发育启动。     

血清抗苗勒管激素水平与多囊卵巢综合征激素特征关系的研究

目的探讨抗苗勒管激素（AMH）与多囊卵巢综合征（PCOS）卵泡发育和激素特征的关系。方法测定47例PCOS患者血清AMH、卵泡刺激素、黄体生成素、催乳素、睾酮、雌二醇水平以及双侧卵巢卵泡数（FN）,并取23例月经周期正常妇女进行对照。结果PCOS组血清AMH（32．57±13．31）pmol／L、T（3．01±0．12）nmol／L、LH（17．68±3．11）U／L水平,明显高于对照组AMH（13．06±4．59）pmol／L、T（1．31±0．02）nmol／L、LH（9．07±2．09）U／L,两组差异有统计学意义（P〈0．01）,并且两组血清AMH水平都与FN呈正相关。结论血清AMH异常可能是PCOS卵泡发育异常和性激素合成失调的原因之一;测定血清AMH水平为诊断和研究PCOS提供了一个重要的方法。     

卵泡液中TGFβ1和性激素水平的测定及临床意义

OBJECTIVE: To investigate the role of transforming growth factor-beta1 (TGFbeta1) and the sex hormones in the follicular fluid (FF) on the day of ovum pick-up (OPU) during controlled ovarian hyperstimulation (COH) cycles. METHODS: FF and the oocytes were obtained from the follicles of 90 women undergoing ovulation stimulation in vitro fertilization (IVF) treatment. TGFbeta1, estradiol, progesterone, and lutropin concentrations in the FF samples collected during transvaginal oocyte retrieval were measured using enzyme-linked immunosorbent assay (ELISA). The maturity and fertilization of the oocytes were observed, and ultrasonography performed to confirm clinical pregnancies 4 weeks after the embryo transfer. RESULTS: In the FF containing mature oocytes, progesterone and lutropin concentrations were significantly higher than those in the FF with immature oocytes. The mean concentrations of TGF beta1, progesterone and lutropin in the FF from fertilized subjects were obviously higher than those in the FF from non-fertilized subjects (P < 0.05), and in subjects with pregnancy, higher mean concentrations of TGFbeta1 and lutropin were detected as compared with the concentrations measured from non-pregnancy subjects (3 631.4+/-1 426.3 pg/ml and 0.74+/-0.25 mIU/ml vs 2 189.2+/-1 180.4 pg/ml and 0.52+/-0.29 mIU/ml respectively, P < 0.05). Estradiol concentrations in the FF seemed to undergo no obvious changes during the whole procedure, and evinced no significant differences between the groups. CONCLUSION: Higher lutropin and progesterone concentrations in the FF on the day of OPU may promote oocyte maturation, while TGFbeta1 and lutropin levels appear to be associated with the maturation and fertilization of the oocytes, and may be indicative of the IVF outcome.     

人胎儿卵泡发育及培养时对促性腺激素反应的形态学研究

取人胎儿卵巢19例作体外培养,观察培养前后与加不同比例促性腺激素后的卵巢形态学变化。培养维持最长达6d。随妊娠进展,胎儿卵泡逐渐生长,自16周起胎儿卵巢在体外就有对促性腺激素起反应的潜力。24周以上的胎儿卵巢可见窦前期和窦期卵泡,伴有透明带基膜和泡膜细胞形成。给予适当剂量的促性腺激素(FSH、LH或hMG)体外培养,可维持培养前的良好状态,不加激素组则易见卵泡闭锁。本文19例胎儿卵巢加激素培养未见发育到较大的窦状卵泡。     

1,6-二磷酸果糖对冻融移植小鼠卵巢组织的影响

目的:在冷冻液中增加能量底物1,6二磷酸果糖(Fructose-1,6-bisphosphate,FDP),观察FDP是否对冻存卵巢有保护作用.将9～12周龄小鼠的卵巢取出,在含(FDP组)或不含FDP(LN2V组)的冷冻液中平衡后快速深低温冻存,解冻后行自体双肾被膜下移植,观察动情周期、移植21 d后取移植物固定行H...