多囊卵巢综合征卵泡膜细胞功能的研究

多囊卵巢综合征(PCOS)是女性最常见的内分泌疾病之一,影响了6%-10%的育龄期妇女。PCOS卵巢的特性是卵泡内膜过度增生。而卵泡膜细胞是PCOS患者雄激素合成过多的原始来源之一。PCOS的卵泡膜细胞中CYP11A1、CYP17、StAR、3β-HSD表达增加。PCOS卵巢雄激素过多的发病机制中存在MAPK信号通路的改变,胰岛素能调控PCOS卵泡膜细胞甾体激素的合成。因此,对于PCOS卵泡膜细胞功能的研究将有助于此疾病的病因学和治疗学的发展。     

卵巢性索间质肿瘤的病理分类激素分泌及临床表现

卵巢性索间质肿瘤的病理分类激素分泌及临床表现天津第二中心医院病理科（３００１２０）范女郎娣天津中心妇产科医院内分泌科（３０００５２）朱楣光１概述［１］卵巢性索－间质肿瘤约占卵巢肿瘤的５％，由性索和胚胎性腺特殊性间质衍化而来，目前倾向于前者衍化为粒层及...     

恶性滋养细胞肿瘤的激素分泌

恶性滋养细胞肿瘤的激素分泌北京协和医院（１００７３０）杨秀玉滋养细胞肿瘤能产生多种激素，一类为糖蛋白激素，如绒毛膜促性腺激素（ｈＣＧ）、促甲状腺激素（ＴＳＨ）、胎盘生乳素（ｈＰＬ）及妊娠特异性糖蛋白（ＳＰ１）；另一类为类固醇激素如雌二醇（Ｅ２）、雌三...     

雄烯二酮、胰岛素和促黄体激素对人类卵巢颗粒黄体细胞产生雌、孕激素的影响

雄烯二酮、胰岛素和促黄体激素(LH)是调节卵巢颗粒细胞产生甾体激素的重要因素,分泌过多及激素间平衡失调会影响卵细胞的成熟。为观察高浓度雄烯二酮、胰岛素和LH对人卵颗粒细胞产生雌、孕激素的调节及三者的相互作用进行此项研究。实验所用人卵巢颗粒细胞是从因输卵管病变接受体外受精的妇女卵泡穿刺获得。将所取颗粒细胞置于     

白藜芦醇调控高雄激素诱导卵巢细胞乙酰化的研究进展

多囊卵巢综合征（polycystic ovarian syndrome，PCOS）是一种以高雄激素表现为典型特征的育龄女性常见内分泌疾病，发病率高，异质性高，病因不明，现阶段临床仍采取对症治疗为主的策略。近年研究显示，PCOS患者卵巢细胞中组蛋白乙酰化修饰异常，与多种组蛋白乙酰转移酶和组蛋白去乙酰化酶的失衡有关。其中沉默信息调节因子1（silent information regulator 1，SIRT1）属组蛋白去乙酰化酶，可被白藜芦醇激活而降低相应组蛋白的乙酰化水平；且白藜芦醇可借助对抗氧化应激、调节雄激素和雌激素水平等途径缓解PCOS相关症状。综述去乙酰化酶SIRT1的激动剂白藜芦醇对高雄诱导的PCOS卵巢异常乙酰化的调控机制，以及白藜芦醇对卵巢功能的修复机制，展望白藜芦醇在PCOS临床治疗的潜在应用前景。     

卵巢颗粒细胞瘤及卵泡膜细胞瘤53例临床分析

卵巢颗粒细胞瘤及卵泡膜细胞瘤５３例临床分析陈亚枫卵巢颗粒细胞瘤及卵泡膜细胞瘤属于卵巢性索间质肿瘤，亦称卵巢功能性肿瘤。常伴有不规则阴道出血及闭经。本研究对我院收治的５３例进行回顾性分析，现将结果报告如下。一、临床资料１．一般情况：我院自１９６４年１月...     

白细胞介素1与多囊卵巢综合征肥胖的相关性

目的 探讨炎症细胞因子——白细胞介素1(IL-1)β亚型(IL-1β)和IL-1受体拮抗剂(IL-1ra)与多囊卵巢综合征(PCOS)患者肥胖发生的关系.方法(1)2006年10月至2007年1月,选择在北京大学第三医院生殖医学中心就诊的PCOS患者118例,根据WHO国际肥胖特别工作组2000年提出的亚太地区肥胖标准[体质指数(BMI)≥25 kg/m2]分为PCOS肥胖组56例,PCOS非肥胖组62例.采用PCR技术检测IL-1β基因-511位点、第5外显子位点和IL-1 ra基因第2内含子的基因多态性.(2)随机抽取PCOS肥胖组患者29例和PCOS非肥胖组患者31例,采用ELISA法检测血清IL-1β、IL-1ra水平,并测定空腹血糖(FBG)、空腹胰岛素(FINS)、超敏C反应蛋白水平和白细胞计数.结果(1)基因型和等位基因频率:PCOS肥胖组患者IL-1β基因-511位点T/T基因型频率及T等位基因频率分别44.6％和63.4％,PCOS非肥胖组患者分别为11.3％和39.5％,两组分别比较,差异均有统计学意义(P＜0.05);PCOS肥胖组患者IL-1 ra基因Ⅰ/Ⅴ基因型频率及Ⅴ等位基因频率分别为19.6％和9.8％,PCOS非肥胖组分别为3.2％和1.6％,两组分别比较,差异也均有统计学意义(P＜0.05).(2)血清学检测结果:PCOS肥胖组患者血清IL-1β水平为(149±36)ng/L,高于PCOS非肥胖组患者的(96±42)ng/L,PCOS肥胖组患者IL-1 ra水平为(284±97)ng/L,高于PCOS非肥胖组患者的(208±84)ng/L,两组分别比较,差异均有统计学意义(P＜0.05).PCOS肥胖组患者FBG、FINS、超敏C反应蛋白水平和白细胞计数分别为(5.1±0.7)mmol/L、(17±9)mU/L、(1.5±0.6)mg/L、(7.0±2.3)×109/L,PCOS非肥胖组患者分别为(4.9±0.5)mmol/L、(11 ±8)mU/L、(0.9 ±0.4)mg/L、(5.9±1.3)×109/L,两组间各项指标分别比较,差异均有统计学意义(P＜0.05).(3)IL与BMI的相关性:PCOS患者血清IL-1β(r=0.673)、IL-1ra(r =0.557)与BMI呈线性正相关(P均＜0.05).结论 炎症因子IL-1β和IL-1ra与PCOS患者肥胖的发生明显相关,IL-1β基因-511位点携带T等位基因和IL-1ra基因携带等位基因Ⅴ的PCOS患者更有发生肥胖的倾向.     

抗菌勒管激素作为卵巢颗粒细胞瘤肿瘤标记物的实验研究

研究抗苗勒管激素（ａｎｔｉ－ｍｕｌｌｅｒｉａｎ ｈｏｒｍｏｎｅ,ＡＭＨ）,在诊断卵巢颗粒细胞瘤（ｇｒａｎｕｌｏｓａ ｃｅｌｌ ｔｕｍｏｒ,ＧＣＴ）中作为特异特性肿瘤标记的实验依据。方法采用免疫组织化学,原位杂交、Ｎｏｔｒｈｅｒｎ杂交法研究转基因小鼠早、晚期卵巢ＧＣＴ中ＡＭＨ及其受体ｍＲＮＡ的表达,并用放射性碘（＾１２５Ｉ）标记法,Ｎｏｒｈｔｅｎ杂交法研究原卵巢ＧＣＴ细胞中ＡＭＨ及其受体ｍＲＮＡ的表     

白细胞介素-1α对卵巢上皮细胞及卵巢癌细胞11β类固醇脱氢酶基因表达的影响

目的探讨白细胞介素1α(IL1α)对正常人卵巢上皮细胞及卵巢癌细胞株11β类固醇脱氢酶(11βHSD)基因表达的影响。方法体外培养人卵巢上皮细胞(HOSE)及卵巢癌细胞株SKOV3、PEO4和PEO14,应用逆转录多聚酶链反应(RTPCR)和实时荧光定量聚合酶链反应(realtimePCR)方法比较上述4种细胞中11β类固醇脱氢酶1(11βHSD1)、11β类固醇脱氢酶2(11βHSD2)和白细胞介素1受体(IL1R)mRNA水平的表达及加入不同浓度的IL1α后,4种细胞中11βHSD1和11βHSD2mRNA表达水平的变化。结果正常卵巢上皮细胞中11βHSD1和IL1RmRNA表达水平较高,11βHSD2mRNA表达水平较低;而卵巢癌细胞株SKOV3和PEO4中11βHSD1和IL1RmRNA表达水平较低,11βHSD2mRNA表达水平较高。加入不同浓度的IL1α后,正常卵巢上皮细胞11βHSD1mRNA表达水平明显升高(P<0.01);PEO14细胞11βHSD1mRNA表达水平也升高(P<0.05);SKOV3和PEO411βHSD1mRNA表达水平无明显变化;正常卵巢上皮细胞11βHSD2mRNA表达水平无明显变化,卵巢癌细胞株PEO4、SKOV3及PEO1411βHSD2mRNA表达水平不同程度升高(P<0.05)。结论卵巢癌细胞丧失了对炎性细胞因子白细胞介素1反应的能力;11β类固醇脱氢酶异构体表达形式的不同是肿瘤细胞转化的一种特征,它可能与卵巢上皮细胞的恶性转化有关。     

一氧化氮及一氧化氮合酶与卵巢肿瘤凋亡的关系

目的探讨卵巢良、恶性肿瘤患者血清、肿瘤组织、腹水中一氧化氮(nitricoxide,NO)及其合酶(nitricoxidesynthase,NOS)含量与肿瘤凋亡的关系。方法用分光光度计测定NO、NOS、iNOS活性,流式细胞仪检测细胞凋亡。结果恶性卵巢肿瘤患者血清、肿瘤组织中NO、NOS、iNOS高于良性肿瘤患者(P<0.05);腹水或腹腔冲洗液中iNOS亦高于良性(P<0.05),但腹水中NO、NOS与良性无差异(P>0.05)。恶性肿瘤细胞凋亡率高于良性(P<0.05)。结论NO、NOS、iNOS与卵巢肿瘤的恶性行为及凋亡有关,检测患者血清、组织、腹水中的NO、NOS、iNOS对鉴别良、恶性肿瘤有一定参考价值,可作为观测卵巢癌疗效及预后的指标。     

黄素化颗粒细胞端粒酶的表达及与卵巢功能关系的初步研究

目的　了解端粒酶在人卵巢黄素化颗粒细胞的表达 ,及与卵巢生殖功能的关系。方法　收集 2 2例行体外受精 胚胎移植或卵胞浆单精子显微注射患者的卵巢黄素化颗粒细胞 ,采用原位杂交法及端粒末端重复序列扩增法 ,分别检测人端粒酶催化亚单位 (hTERT)mRNA及端粒酶活性。结果　黄素化颗粒细胞普遍表达hTERTmRNA ,但其表达强弱不一。 2 2份卵巢黄素化颗粒细胞标本中 ,16份 (73% )有端粒酶活性 ,与血清基础促卵泡激素水平呈负相关 (P <0 .0 1) ,且随年龄增长呈下降趋势。结论　端粒酶可能对维持卵巢功能起重要作用 ,卵巢功能下降可能与卵巢颗粒细胞端粒酶活性降低有关     

瘦素对人卵巢黄素化颗粒细胞雌、孕激素分泌的体外实验研究

目的探讨瘦素在体外对人卵巢颗粒细胞雌激素和孕激素生成的影响。方法将来自体外受精-胚胎移植（IVF-ET）的卵巢黄素化颗粒细胞纯化后,在不同浓度瘦素(0、10、30、100、300ng/ml)和人绝经期促性腺激素（hMG,0、0.1、0.2、0.5、1、2、5、10IU/ml）单独或联合作用下进行体外培养,收集培养液,采用放射免疫方法测定颗粒细胞产生雌二醇及孕酮的量。结果不同浓度的瘦素对雌二醇、孕酮的生成无影响;hMG为5IU/ml时,雌二醇水平平均为2.36×10-11mol/L,加入不同浓度瘦素（10、30、100、300ng/ml）,雌二醇的生成均受到明显的抑制,随瘦素浓度增加,雌二醇水平逐渐降低（P<0.05）,孕酮水平无明显变化;hMG浓度小于0.5IU/ml时,瘦素对雌二醇生成无影响;hMG浓度大于0.5IU/ml、瘦素为100ng/ml时,雌二醇水平低于不加瘦素的对照（P<0.05）。结论瘦素参与卵巢颗粒细胞激素生成的调节,在卵泡发育和黄体形成中有一定的作用。     

妊娠高血压综合征患者血浆P-选择素和一氧化氮含量变化及意义

目的　探讨妊娠高血压综合征（妊高征）患者血浆P-选择素和一氧化氮(nitricoxide,NO)含量的变化及其在妊高征发病中的意义。方法　采用酶联免疫吸附法测定36例妊高征患者（妊高征组,其中轻度妊高征患者11例,中重度妊高征患者25例）、18例正常晚孕妇女（正常晚孕组）和19例正常非孕妇女（正常非孕组）血浆P-选择素含量。用镀铜镉还原法测定血浆中代表NO水平的亚硝酸盐及硝酸盐（nitrite and nitrate,NO     

AMH蛋白在性成熟小鼠卵泡发育不同阶段及自然衰老过程卵巢组织中的表达变化

目的:通过检测抗苗勒氏管激素(AMH)蛋白在成年小鼠卵泡发育不同阶段和自然衰老过程卵巢中的表达变化,为卵泡的生长发育调控及卵巢衰老发生发展的机制研究奠定理论基础。方法:选用12,24,36,48周龄自然衰老C57BL/6小鼠为模型。收集12周龄C57BL/6小鼠不同发育阶段的卵巢和24,36,48周龄自然衰老C57BL/6小鼠卵巢。采用免疫组织化学方法和蛋白印迹技术检测各组卵巢中AMH蛋白的表达。结果:免疫组织化学显示,AMH蛋白在始基卵泡、初级卵泡,次级卵泡、窦状卵泡的颗粒细胞中均有表达,但在黄体和闭锁卵泡的颗粒细胞中未见表达。始基卵泡颗粒细胞中AMH蛋白表达明显低于初级卵泡、次级卵泡和窦状卵泡(P<0.05);窦状卵泡颗粒细胞中AMH蛋白表达明显低于初级卵泡和次级卵泡(P<0.05);初级卵泡和次级卵泡的AMH蛋白表达未见明显差异(P>0.05)。不同鼠龄的小鼠同级别卵泡颗粒细胞AMH的染色程度未见明显差异;但是24周龄以后间质中开始出现AMH蛋白表达,并且AMH的表达随鼠龄增加而上升;48周龄小鼠卵巢间质中AMH蛋白表达明显高于24周龄和36周龄(P<0.05);24周龄与36周龄比较差异无统计学意义(P>0.05)。蛋白印迹技术结果显示,AMH蛋白在小鼠卵巢组织中的表达随鼠龄增加而降低,差异有统计学意义(P<0.05)。结论:AMH蛋白主要表达于卵泡发育早期阶段的颗粒细胞,并且其在间质中随鼠龄增加表达增强,但是在整个卵巢组织中其表达量随鼠龄增加而降低。提示AMH可能在卵泡发育以及卵巢衰老中起重要的调控作用。     

尿皮素对体外培养的人脐静脉内皮细胞一氧化氮及一氧化氮合酶的影响

目的:通过检测尿皮素(UCN)对人脐静脉内皮细胞(HUVEC)一氧化氮(NO)生成及一氧化氮合酶(NOS)表达的影响,探讨UCN调节胎儿-胎盘血管张力的分子机制。方法:在离体培养的HUVEC中加入不同浓度的UCN(0、0.1、1、10nmol/L)、10nmol/L促肾上腺皮质激素释放激素(CRH)及向以上各组UCN/CRH同时加α-helical-CRH进行体外培养,用硝酸还原酶法检测细胞上清液中NO的水平,蛋白印迹法检测内皮型一氧化氮合酶(eNOS)和诱导型一氧化氮合酶(iNOS)蛋白表达水平的变化。结果:在与UCN共同培养4~8h后,HUVEC细胞上清液中NO水平呈时间和剂量依赖性变化,随着培养时间的延长,UCN浓度增加,NO水平逐渐升高;UCN组HUVEC iNOS表达水平比对照组明显升高(P<0.05),且呈浓度依赖性升高。而各组eNOS表达水平与对照组比较无明显变化(P>0.05)。UCN/CRH α-helical-CRH组HUVEC上清液中NO水平和HU-VEC iNOS表达水平与对照组比较无明显变化,差异无统计学意义(P>0.05),与相同浓度UCN/CRH组比较NO水平和iNOS蛋白表达水平明显降低,差异有统计学意义(P<0.05)。结论:UCN可促HUVEC合成及释放NO,NO生成增多与UCN通过促进肾上腺皮质激素释放激素2β受体(CRH-R2β)正相调节iNOS蛋白表达有关,这可能是UCN调节胎儿-胎盘血管张力的分子机制之一。     

Involvement of nitric oxide and the ovarian blood follicle barrier in murine follicular cyst development

OBJECTIVE: To test the involvement of nitric oxide in murine ovarian follicular cysts. DESIGN: Controlled animal study. SETTING: Academic research environment. ANIMAL(S): Immature female B6D2F1 mice at 23 +/- 2 days old.Ovarian cysts were induced by implanting miniosmotic pumps that delivered and maintained constant levels of hCG. Nitric oxide studies included the delivery of nitric oxide synthase (NOS) inhibitors, N(G)-nitro-L-arginine methyl ester (L-NAME), or N(G)-nitro-D-arginine methyl ester, by the same method.Ovulation assays measured cumulus oocyte complexes and blood follicle barrier (BFB) function. RESULT(S): Chronic treatment with hCG induced enlarged ovaries containing multiple follicular cysts, which were approximately double the size of follicles in sham-operated mice. These cysts enclosed few, if any granulosa cells, secreted high levels of testosterone, and had impaired ovarian BFB function. Inhibition of NOS by L-NAME during ovarian cyst formation reduced the size of follicular cysts, sustained normal testosterone levels, and maintained hormonal BFB reactivity in cystic follicles. CONCLUSION(S): Nitric oxide was found to be involved in the formation of hCG-induced murine follicular cysts and complications associated with these cysts were ameliorated by the NOS inhibitor L-NAME.     

Dual effect of anandamide on rat placenta nitric oxide synthesis

Anandamide (AEA) has been reported to have pleiotropic effects on reproduction, but the mechanism by which it exerts these effects is unclear. The aim of this study is to characterize rat placental endocannabinoid system and to analyze the possible functional role of AEA in the regulation of NO levels in rat placenta during pregnancy. We found that cannabinoids receptors (CB1 and CB2), FAAH and TRPV1 were expressed in chorio-allantoic placenta. NOS activity peaked at day 13 and decreased with progression of pregnancy. Both exogenous and endogenous AEA significantly decreased NOS activity. Although pre-incubation with AM251 (CB1 antagonist) or AM630 (CB2 antagonist) had no effect, co-incubation with both antagonists induced NOS activity. Furthermore, pre-incubation with exogenous AEA and both antagonists resulted in the induction of placental NOS activity and this effect was reverted with capsazepine (selective TRPV1 antagonist). Additionally, the enhanced NO synthesis caused by capsaicin was abrogated by co-treatment with capsazepine, illustrating that NOS activity could be modulated by TRPV1. Finally, the inhibition of TRPV1 receptor by capsazepine caused a significant fall in NOS activity. These data support the concept that AEA modulates NO levels by two independent pathways: (1) diminishing the NOS activity via CBs; and (2) stimulating NO synthesis via TRPV1. We hypothesized that AEA have an important implication in the normal function of placental tissues.     

Stimulated nitric oxide release and nitric oxide sensitivity in forearm arterial vasculature during normotensive and preeclamptic pregnancy

OBJECTIVE: We sought to determine whether the enhanced forearm vascular activity of nitric oxide during pregnancy and preeclampsia is associated with altered smooth muscle sensitivity to nitric oxide or with stimulated nitric oxide release. STUDY DESIGN: Forearm blood flow responses to brachial artery infusion of glyceryl trinitrate (a nitric oxide donor), serotonin (an endothelium-dependent nitric oxide-mediated agonist), and ritodrine (a beta-adrenergic receptor agonist) were studied in 10 nonpregnant women, 12 pregnant women, and 7 women with preeclampsia by means of strain-gauge plethysmography. Responses to each drug (maximum dilator response and the sum of the percentage of dilator responses to each drug) were compared by analysis of variance. RESULTS: Compared with nonpregnant women, pregnant subjects showed reduced responses to serotonin (summary response, 117 +/- 19 vs 221 +/- 30; P <.05). Responses to serotonin were reduced in the group with preeclampsia compared with those in the nonpregnant group (summary response, 71 +/- 28; P <.05) but did not differ from the responses in pregnant women. There were no differences between responses to glyceryl trinitrate and responses to ritodrine in any of the groups. CONCLUSION: Vascular smooth muscle sensitivity to nitric oxide is not altered in normal pregnancy or preeclampsia, but dilator responses to serotonin appear blunted. Alterations in serotonin receptor coupling to nitric oxide synthase, or a limitation of availability of the substrate for nitric oxide synthase (L-arginine) during pregnancy, could account for the reduction in stimulated nitric oxide release.     

Differential in vitro actions of nitric oxide on human endometrial cell survival

Objective

To investigate the presence of caspase-3 and Bcl-2 concentration in human endometrial tissue throughout the menstrual cycle, and study the effect of nitric oxide (NO) on cell proliferation and apoptosis during culture.

Design

Expression of caspase-3 and Bcl-2 concentration in endometrial explants, and examination of L-arginine (L-Arg) effect on epithelial and stromal cell proliferation and apoptosis in vitro.

Setting

Prospective study.

Patient(s)

Twenty-seven eumenorrheic women (37 ± 1.2 years).

Intervention(s)

Endometrial samples were obtained with Pipelle suction curette from the corpus of the uterus.

Main outcome measure(s)

Apoptosis (annexin V-FITC binding), Bcl-2 concentration (ELISA), caspase-3 (immunohistochemistry), cell proliferation (spectrophotometric assay), and gene expression (RT-PCR).

Result(s)

Caspase-3 was detected by immunoassay in epithelial tissue throughout the menstrual cycle and in stroma during secretory phase. The Bcl-2 concentration was similar in endometrial homogenates obtained throughout the menstrual cycle, but L-Arg decreased Bcl-2 only in endometrium from the proliferative phase. In epithelial cells, NO increased apoptosis by 2.1 ± 0.2-fold, augmented mRNA expression of Bax, and reduced expression of Bcl-2 compared with basal cultures. In stromal cells, NO increased cell proliferation in a dose-dependent manner, an effect that was blocked by a NO synthase inhibitor.

Conclusion(s)

These data indicate that NO has a differential regulatory function on endometrial cell survival, as indicated by the results on stromal cell proliferation and epithelial cell apoptosis during culture, which suggests paracrine interactions between both cell types.