控制性超促排卵对黄体期子宫内膜胶原纤维表达的影响

目的:探讨IVF-ET妇女控制性超促排卵(COH)时着床窗期子宫内膜胶原纤维(collagen fibril,CF)对子宫内膜容受性(ER)的影响。方法:共对25个刺激周期(SC)和20个自然周期(NC)妇女分别在取卵后或排卵后的第7±1日行外周血雌、孕激素水平检测和子宫内膜活检术,常规HE染色确定组织学分期,采用透射电子显微镜观察CF增生情况和改良Masson染色进行CF定量分析。结果:SC组外周血雌、孕激素水平均显著高于NC组(P<0.05)。SC组分泌期较NC组提前1～2 d,内膜腺体扩张和间质水肿明显。SC组分泌早、中和晚期内膜间质中的CF较NC组显著升高(P<0.01,P<0.05和P<0.05),且SC组分泌早期SC明显高于分泌中期(P<0.05)。结论:COH过程中过高的雌、孕激素水平可使子宫内膜分泌期提前并刺激内膜CF的过度增生,这可能是导致COH周期ER下降和着床失败的原因。     

促排卵周期PCOS妇女子宫内膜纤维粘连蛋白及层粘连蛋白的表达

目的:了解促排卵药物氯米酚(CC)、hMG及GnRH-a对黄体中期子宫内膜内膜纤维粘连 蛋白(FN)及层粘连蛋白(LN)表达的影响。方法:应用单克隆抗体,采用免疫组织化学技术检测50 例正常妇女自然周期以及50例正常妇女,45例多囊卵巢综合征妇女应用CC／hCG,CC／hMG／hCG 及GnRH-a／hMG／hCG方案促排卵治疗后黄体中期子宫内膜FN和LN的表达。结果:子宫内膜FN 和LN表达在正常妇女自然周期着床窗口时呈现强阳性;而CC、hMG抑制FN和LN的表达,使 其阳性强度减弱,有显著性统计学差异P<0．01;GnRH-a对FN和LN抑制不明显。同时妊娠者较 未妊娠者FN和LN表达强度高。结论:CC／hCG及CC／hMG／hCG方案促排卵后黄体中期子宫内膜 中FN和LN表达下降或缺失,内膜容受性下降,妊娠率降低。     

米非司酮治疗子宫内膜增生症前后ER和AR的表达及意义

目的:探讨米非司酮抑制子宫内膜增生的机制。方法:37例子宫内膜增生症(不伴非典型增生)患者,连续口服米非司酮(剂量10mg/d)治疗3个月,刮取治疗前、后子宫内膜组织,采用免疫组织化学二步法分别检测其ER和AR的表达进行自身对比;另取10例正常增殖期子宫内膜标本作为正常对照组进行比较。结果:子宫内膜增生症治疗前ER表达、AR表达均显著高于正常对照组(P<0.05),ER在腺体和间质细胞中均有表达,AR主要表达于间质细胞,腺体中几乎无表达。米非司酮治疗后腺体和间质ER表达比治疗前均下降(P<0.05),而间质和腺体中AR表达均比治疗前增加(P<0.05)。结论:米非司酮可通过下调ER和上调AR,从而抑制子宫内膜增生。     

瘦素及瘦素长受体在多囊卵巢综合征患者子宫内膜的表达

目的　探讨瘦素 (leptin)和瘦素长受体 (Ob RL)mRNA及蛋白在多囊卵巢综合征(PCOS)患者着床期子宫内膜的表达及意义。方法　应用免疫组织化学技术及原位杂交技术 ,分别对15例正常妇女 (对照组 )、14例无排卵PCOS患者 (PCOS组 )促排卵治疗后着床期子宫内膜的leptin、Ob RL mRNA及蛋白进行测定。结果　对照组中 ,子宫内膜 10例呈分泌期中期改变 ,5例呈分泌期早期改变。PCOS组中 ,子宫内膜 3例呈分泌期中期改变 ,6例呈分泌期早期改变 ,1例呈增殖期改变 ,4例呈增殖期并分泌期早期改变 ;子宫内膜发育不良发生率 79% (11/ 14 ) ,与对照组比较 ,差异有显著性 (P <0 .0 5 )。Ob RL mRNA和蛋白在PCOS患者子宫内膜腺体呈阴性到强阳性表达 ,但多呈弱阳性和阳性表达 ,对照组则呈弱阳性到强阳性表达 ,两组比较 ,差异有显著性 (P <0 .0 5 ) ;leptinmRNA和蛋白在PCOS患者子宫内膜的表达与对照组相似 ,两组比较 ,差异无显著性 (P >0 .0 5 )。结论　促排卵周期中 ,Ob RL 在PCOS患者着床期子宫内膜腺体表达减弱 ,可能与PCOS患者妊娠率低有关     

子宫内膜癌组织中环氧化酶-2增强表达的临床意义

目的:探讨环氧化酶2(COX2)在子宫内膜癌组织中的表达,COX2与雌激素受体(ER)表达的关系及COX2在子宫内膜癌发生发展中的意义。方法:应用RTPCR技术检测30例子宫内膜癌、相应癌旁正常内膜腺体及15例子宫良性肿瘤中COX2mRNA的表达。应用免疫组化方法测定30例内膜癌中ER表达情况。结果:COX2mRNA在30例子宫内膜癌组织中有28例表达水平明显增高,癌组织与正常内膜腺体或良性肿瘤间COX2的表达差异有显著性意义(P<0.05)。ER阳性组与ER阴性组的COX2表达差异有非常显著性意义(P<0.01)。结论:子宫内膜癌组织中COX2mRNA表达水平高于正常内膜腺体或子宫良性肿瘤,激素依赖型子宫内膜癌中COX2表达高于非激素依赖型内膜癌且其表达在子宫内膜癌的发生、发展中具有重要意义。     

类固醇受体辅助活化因子-1和核受体辅阻遏子在子宫腺肌病中的表达

目的:通过检测子宫腺肌病组织芯片中雌激素受体(ER)、孕激素受体(PR)、类固醇受体辅助活化因子-1(SRC-1)和核受体辅阻遏子(NCoR)的表达,观察SRC-1、NCoR与ER、PR之间的关系,探讨其在子宫腺肌病发生、发展中的作用。方法:用组织芯片技术和免疫组化方法检测42例子宫腺肌病患者异位子宫内膜组织(异位内膜组)和15例因子宫肌瘤行子宫切除术患者非瘤区对照子宫内膜组织(对照内膜组)中ER、PR、SRC-1和NCoR的阳性细胞的平均光密度值(IOD),及两组子宫内膜组织的增殖期和分泌期的IOD值,并进行比较。结果:ER、PR在异位内膜组中的IOD值显著低于对照内膜组(P<0.05);SRC-1在异位内膜组中IOD值明显高于对照内膜组(P<0.05);NCoR在异位内膜组中IOD值明显低于对照内膜组(P<0.05);异位内膜组中SRC-1、NCoR、ER和PR的IOD值增殖期和分泌期的比较,差异无统计学意义(P>0.05);对照内膜组中SRC-1、NCoR、ER和PR的IOD值增殖期高于分泌期(P<0.05)。结论:低表达的ER存在时,SRC-1在异位内膜组织中表达增高,低表达的PR可能降低NCoR的表达,SRC和NCoR可能导致子宫腺肌病异位内膜组织增生活性增高,可能与子宫腺肌病的种植和侵袭有关。     

米非司酮对离体异位与在位子宫内膜雌、孕激素受体含量的影响

目的　探讨子宫内膜异位症(内异症)的异位与在位内膜雌、孕激素受体(ER、PR)含量,及米非司酮对其影响。方法　采用免疫细胞化学法,分析22例内异症患者的在位内膜细胞和其中12例患者的异位内膜细胞体外培养后的ER、PR含量,观察不同浓度米非司酮(1×10-6mol/L和1×10-4mol/L)作用后的变化,并以13例正常子宫内膜作对照。结果　内异症的在位内膜ER、PR含量呈明显周期性变化,分泌早期腺体PR含量显著高于正常子宫内膜［组织化学评分(下同)为2.77±0.32与2.20±0.26,P<0.05］。内异症的异位内膜,增殖期ER(腺体0.65～2.17,间质0.45～1.03)、PR含量(腺体0.55～1.77,间质0.40～1.27)显著低于在位内膜(ER腺体1.50～3.23,间质0.80～1.96;PR腺体1.55～3.34,间质0.98～2.50,P<0.05～0.01);分泌早期无差异;分泌晚期腺体ER含量(3.27±0.31)、PR含量(3.33±0.23)与间质ER含量(1.87±0.31)显著高于在位内膜(分别为0.28±0.11、0.36±0.23和0.26±0.15,P<0.01),而间质PR含量无差异。米非司酮可明显降低内异症的异位和在位内膜ER、PR含量(P<0.01),且米非司酮浓度越高,ER、PR含量降低越明显。结论　内异症的异位和在位内膜ER、PR含量明显不同,米非司酮可下调异位和在位内膜ER、PR的含量。     

GnRH-a控制性超促排卵(COH)对小鼠着床期Hoxa-10基因表达的影响

目的:研究GnRH-a长方案控制性超促排卵(COH)对小鼠妊娠率、胚胎着床率及着床期HOXA-10基因表达的影响,探讨COH影响胚胎着床的机制。方法:实验动物随机分为实验组(GnRH-a+hMG+hCG)和对照组(生理盐水),雌、雄合笼后观察小鼠阴栓率,于着床期取小鼠子宫,观察妊娠率及胚胎着床数,计算着床率;应用RT-PCR、Real-time PCR方法及免疫组织化学法分别检测小鼠着床期子宫内膜Hoxa-10 mRNA及蛋白的表达。结果:实验组小鼠阴栓率及妊娠率均明显低于对照组(P<0.01),实验组胚胎着床数高于对照组(P<0.01),但胚胎着床率较对照组显著降低(P<0.05),着床期小鼠子宫内膜Hoxa-10mRNA及蛋白表达下降,与对照组比较均有明显差异(P<0.05)。结论:GnRH-a长方案COH可引起小鼠着床期子宫内膜Hoxa-10表达下降,导致子宫内膜容受性降低,从而影响小鼠妊娠及胚胎着床。     

GnRHa超排卵对小鼠子宫内膜形态学和雌、孕激素受体表达的影响

探讨 Gn RHa长周期辅助超排卵对小鼠分泌期子宫内膜组织形态和性激素受体表达的影响。方法 :将 40只小鼠随机分成 4组 ,其中 2组给 Gn RHa+h MG+h CG为用药组 ,另 2组为对照组给等体积生理盐水 ,在促排卵和排卵后 2 4、48h利用多功能图象分析仪定量测定子宫内膜厚度、腺体和腺腔面积、周长、最大直径、腺上皮细胞总面积和平均高度 ;SP免疫组化法定量测定腺上皮细胞雌、孕激素受体表达。结果 :( 1 )用药组在注 h CG后 2 4、48h子宫内膜厚度与对照组相比差异无显著性 ( P>0 .0 5 )。 ( 2 )用药组在注 h CG后此两时段腺体和腺腔面积、周长、最大直径以及腺上皮细胞总面积和平均直径均显著小于对照组 ( P均 <0 .0 5 )。同时用药组部分内膜间质和腺体发育不同步。( 3 )用药组在注 h CG后两时段内膜腺上皮细胞 ER、PR免疫组化阳性率和染色强度均显著低于对照组同期值 ( P均 <0 .0 5 )。结论 :Gn RHa辅助超排卵对小鼠子宫内膜组织结构和 ER、PR表达有不利影响 ,提示该方案并不能完全促进生理状态的子宫内膜形成 ,可能为低妊娠率原因之一     

自然及促排卵周期子宫内膜整合素α_4β_1的表达

目的　了解氯米芬 (CC)、绝经期促性腺激素 (hMG)对黄体中期子宫内膜整合素α4 β1表达的影响。方法　应用单克隆抗体 ,采用免疫组织化学技术检测 48例正常妇女自然周期以及 48例正常妇女、30例多囊卵巢综合征患者应用CC/绒毛膜促性腺激素 (hCG)及CC/hMG/hCG方案促排卵治疗后黄体中期子宫内膜整合素α4 β1的表达。结果　子宫内膜整合素α4 β1在正常妇女自然周期着床窗口期呈现强阳性表达 ,而CC、hMG抑制整合素α4 β1的表达 ,两者比较 ,差异有极显著性 (P <0 0 1) ;妊娠者较未妊娠者整合素α4 β1表达强度高。结论　促排卵周期黄体中期整合素α4 β1表达下降或缺失 ,子宫内膜容受性下降 ,妊娠率降低。     

Serum hormone and steroid hormone receptor levels during luteal-phase and long-term treatment with danazol

This study was designed to clarify the effects of danazol on levels of serum luteinizing hormone (LH), follicle-stimulating hormone (FSH), prolactin, progesterone (P), and 17 beta-estradiol and endometrial steroid receptors (for estrogen [ER], progestin [PR], and androgen [AR] ) during luteal-phase and long-term treatment. These levels were compared with midluteal-phase levels for a histologically in-phase endometrium. Danazol given during the luteal phase to patients with in-phase endometrium decreased endometrial steroid receptor levels (total ER and total PR), and decreased serum P, LH, and FSH levels. Ten of the 17 patients treated (59%) still had in-phase endometrium. Danazol (400 mg/day) given for 1 month or more to patients with pelvic endometriosis increased serum LH and FSH levels within the normal range and endometrial total ER and PR levels. It appears that the effects of short-term and long-term treatment with danazol on serum hormone and endometrial steroid receptor levels differ.     

Apoptosis-related proteins and steroid hormone receptors in normal, hyperplastic, and neoplastic endometrium.

The purpose of this study was to evaluate the distribution and frequency of apoptosis-related proteins and their correlation with estrogen, progesterone, and androgen receptors in endometrial tissues. Immunohistochemical analyses of bcl-2, bax, bcl-x, and steroid receptors were performed in 22 endometrial carcinomas, 26 endometrial hyperplasias, and 19 cases of normal cyclical endometrium. Bcl-2 was expressed in 45.4% of carcinomas and 92.3% of hyperplasias. Bax immunostaining was found in 90.9% of carcinomas and 76.9% of hyperplasias. Bcl-x positivity was similar in carcinomas (68.1%) and endometrial hyperplasias (76.9%). In normal cyclical endometria, bcl-2 staining was intense and diffuse in the proliferative phase, but decreased dramatically in the early and mid-secretory phase to reappear in the late secretory phase. Bax was expressed throughout the menstrual cycle but more strongly in the secretory phase. Bcl-x displayed a similar degree of expression in proliferative and secretory endometria. Nineteen carcinomas (86.3%), 25 hyperplasias (96.1%), and 18 normal cyclical endometria (94.7%) were positive for estrogen receptor (ER). Progesterone receptor (PR) was observed in 20 carcinomas (90.9%), all hyperplasias (100%), and 18 normal cyclical endometria (94.7%). Androgen receptor (AR) positivity was seen in 7 carcinomas (31.8%), 6 hyperplasias (23.0%), and 3 normal cyclical endometria (15.7%). There was a statistically positive correlation between bcl-x and ER and a tendency toward significant correlation between bcl-x and PR and between ER and PR in carcinomas. In hyperplasias, there was a significant positive correlation between bcl-2 and PR and between bcl-2 and bax and a negative correlation between ER and bax. There was a statistically significant difference for bcl-2 (p = 0.001) and bax (p = 0.001) between the hyperplasia and carcinoma groups. There was increased expression of bax, decreased expression of bcl-2, and persistence of bcl-x protein in advanced endometrial carcinomas. Our findings show that ovarian hormones have a regulatory role on bcl-2 protein and that there is a correlation between other members of the bcl-2 family (bcl-x and bax) and steroid hormone receptors. Bax/bcl-x may be the major control mechanisms of apoptosis in advanced carcinomas; other members of the bcl-2 family may also be under hormonal control.     

Endometrial steroid receptor during normal menstrual cycle and physiopathology of nidation

Human uterine endometrial steroid receptor [estrogen (ER), progestin (PR) and androgen (AR) receptors] in the normal menstrual cycle were determined at the cellular level. Endometria with pathological out-of-phase findings or with hypoplastic uterus were evaluated for the concentration of steroid receptors, for a better understanding of their pathophysiology. ER and PR levels gradually rose toward the early secretory phase, remained at the maximum level during the mid-secretory phase and dropped to the lowest level during the late secretory phase. Especially nuclear PR reached the maximum level during the mid-secretory phase. These results indicate that the endometrial ER and PR, especially the endometrial nuclear PR, is kept in the maximum level at implantation. AR showed only a slight change in a very low level throughout the menstrual cycle. This indicates that androgen may not play so important a role in endometrial physiology. Endometria with out-of-phase or with hypoplastic uterus contained significantly low levels of endometrial ER and PR, while serum hormones levels were normal. This indicates indigenous reduction of ER and PR synthesis in such endometria.     

Endometrial hormone receptors in women with dysfunctional uterine bleeding

Objective The objective was to study the estrogen receptor (ER) and progesterone receptor (PR) expression in endometrium of women with dysfunctional uterine bleeding (DUB) as compared to women with normal menstrual cycles.Methods In this study, 30 patients and 20 controls were selected. Transvaginal ultrasound and endometrial sampling for histology and ER and PR estimation immunohistochemically was carried out for all the subjects. Students t-test and linear correlation was used for statistical analysis. Their response to treatment was assessed by clinical follow-up.Results Endometrial thickness and ER and PR levels in DUB patients were significantly higher. In cases showing hyperplastic endometria, ER and PR levels were higher than patients with normal histology. In contrast to hyperplastic tissue, steroid receptor levels decrease in hyperplastic tissue containing atypia.Conclusion Altered endometrial morphology and increased receptor levels in DUB patients suggest that unopposed estrogen effect could have an important role in the pathogenesis of DUB. Cases of DUB, which showed atypical hyperplasia, may have a down-regulation of these receptors and could be a precursor lesion to carcinoma and thus do not respond to medical therapy by hormones.     

Norplant皮下埋植后子宫内膜形态和雌、孕激素受体含量的变化

目的探讨Ｎｏｒｐｌａｎｔ皮下埋植后子宫内膜形态和雌、孕激素受体（ＥＲ、ＰＲ）含量的变化与子宫异常出血的关系。方法应用免疫组化、ＷｅｓｔｅｒｎＢｌｏｔ和原位杂交技术结合计算机图像分析,观察１６例Ｎｏｒｐｌａｎｔ皮下埋植后和２３例正常周期的内膜的形态学改变、细胞增殖状态及ＥＲ、ＰＲ蛋白及其ｍＲＮＡ的表达。结果埋植Ｎｏｒｐｌａｎｔ后,内膜ＤＮＡ含量降低;螺旋动脉数量减少,腺体的构成比例降低;腺体构成比例与雌二醇（Ｅ２）水平呈正相关,与子宫异常出血呈负相关,与年龄和体重无相关性,随使用期延长,腺体构成比增加。埋植后,内膜ＥＲ含量低于月经周期各期水平,但ＰＲ含量相当于或高于月经周期最高水平。ＥＲ、ＰＲｍＲＮＡ表达减弱,以ＥＲｍＲＮＡ表达降低更明显。结论内膜腺体和螺旋动脉再生修复不良和ＥＲ、ＰＲ表达异常,可能是Ｎｏｒｐｌａｎｔ致子宫异常出血的病理学基础     

Androgen, estrogen and progesterone receptor expression in the human uterus during the menstrual cycle

Cyclic changes in steroid receptor expression in endometrial cells are considered a reflection of its differential functions. Besides estrogen and progestogens, androgens have also been suggested to affect the biological function of the female reproductive tract. We investigated the distribution and intensity of immuno-cytochemical estrogen receptor (ER), progesterone receptor (PR) and androgen receptor (AR) staining in the various cell types of human endometrium and myometrium during the different menstrual cycle phases in 30 paraffin-embedded sections.AR expression in endometrial stromal cells decreased gradually from early proliferative till mid secretory phase. In the late secretory phase, AR expression in all cell types distinguished. Staining of epithelial cells was moderate. The disappearance of AR expression before cyclic separation of endometrial tissue may be causally related or just an epiphenomenon. Due to local competition for 5alpha-reduction of testosterone and the excess of progesterone in the secretory phase, the level of dihydrotestosterone (DHT) will be diminished. Hypothetically, if AR synthesis in endometrium would be DHT-dependent, it would explain the lack of AR expression in the late secretory phase.     

Analysis of estrogen- and progesterone-receptor expression in endometrial polyps

Study objectiveTo investigate the presence of estrogen receptors (ER) and progesterone receptors (PR) in the glandular epithelium and stroma of endometrial polyps in women who underwent hysteroscopic polypectomy.DesignProspective study (Canadian Task Force classification II-3.SettingHospital de Servidor Público Estadual de São Paulo “Francisco Morato de Oliveira.”PatientsForty-eight patients with a solitary endometrial polyp who underwent hysteroscopic polypectomy and resection of an endometrial fragment.InterventionThe estrogen and progesterone receptor expression, in the polyp, was compared with the endometrial hormone expression of the same patients.Measurements and main resultsThe specimens were analyzed by immunohistochemistry. The percentage of staining cells was determined as follows: grade I, 0% to 25%; grade II, 26% to 50%; grade III, 51% to 75%; and grade IV, 76% to 100% of stained nuclei. The intensity of nuclear staining was classified as grade I, absent; grade II, weak; grade III, strong; and grade IV, very strong staining. The sum of both grades was the histochemical score. The total scores of polyp and endometrium were statistically compared. The total scores of ER of glandular epithelium were 258 in polyp and 211 in endometrium. As to stroma, it was 155 in polyp and 163 in endometrium. The total scores of PR of glandular epithelium were 286 in polyp and 211 in endometrium. As to stroma, the totals were 76 in polyp and 77 in endometrium. In immunohistochemistry, the concentrations of both ER and PR in glandular epithelium were significantly higher in endometrial polyp than in normal endometrium. The concentrations of ER and PR in the stroma were similar in the polyp and endometrium. The concentrations of these receptors in the glandular epithelium and stroma were similar in postmenopausal and premenopausal patients.ConclusionThe concentrations of ER and PR in glandular epithelium were significantly higher in endometrial polyp than in normal endometrium. The concentrations of these receptors in the glandular epithelium and stroma were similar in the postmenopausal and pre-menopausal patients.     

PELP1在Ⅱ型子宫内膜癌组织中的表达及其与ER相关性研究

目的:探讨脯氨酸、谷氨酸和亮氨酸富集蛋白1(PELP1)在II型子宫内膜癌中的表达及其与雌激素受体(ER)的相关性。方法:免疫组化法检测84例子宫内膜癌及40例正常子宫内膜组织中PELP1和ER表达水平并对其进行相关性分析。结果:I型子宫内膜癌及增殖期子宫内膜组织中PELP1、ER阳性表达率高于II型子宫内膜癌及分泌期子宫内膜组织,差异均有统计学意义(P0.05);II型子宫内膜癌与分泌期子宫内膜组织比较,差异均无统计学差异(P0.05)。Pearson相关分析提示,PELP1与ER在I型子宫内膜癌组织中表达量呈正相关(rs=0.348,P=0.01),而在II型子宫内膜癌组织中无相关性(rs=0.268,P=0.144)。结论:PELP1在II型子宫内膜癌组织中的表达较少,且与ER表达无明显相关性。     

长期应用小剂量米非司酮对子宫内膜形态和雌、孕激素受体的影响

本文利用HE染色和免疫组化方法结合计算机辅助图象分析技术观察了长期服用小剂量米非司酮对子宫内膜形态和雌、孕激素受体（ER、PR）的影响，15例子宫肌瘤患者于早卵泡期服用来非司酮（10例，10mg／日；5例，20mg／日）治疗，1～3个月后取内膜，另10例肌瘤患者取内膜作对照。结果：用药后所有的患者均闭经，内膜变薄，发育不良，无周期性改变，大部分基质致密，基质与腺体发育不同步，同时相间存在增殖与分泌期样改变，以增殖期样改变为主，腺体以分泌期样改变为主，腺体大小形态不一，但分泌不良。免疫组化显示内膜ER、PR明显分布不均，图象分析结果表明用药后内膜ER、PR相当于对照组增殖早中期水平。结果表明每日10mg米非司酮可明显抑制排卵并影响内膜成熟，在无孕激素存在的情况下，米非司酮有微弱的孕激素样作用。