月经周期人子宫内膜腺上皮和基质细胞HOXA11表达的差异

目的:探讨HOXA11在月经周期人子宫内膜腺上皮和基质细胞中的表达规律及其生理意义。方法:免疫组织化学方法观察38例子宫内膜腺上皮和基质细胞HOXA11蛋白质的表达;用细胞分离筛选法,分离出子宫内膜腺上皮细胞和基质细胞,采用半定量RT-PCR和Dot-blot方法分别观察HOXA11在腺上皮和基质细胞中的表达。结果:腺上皮细胞HOXA11在分泌中晚期表达量较增生期和分泌早期显著降低;基质细胞HOXA11的表达从增生期到分泌期逐渐增加,以分泌中晚期表达量最高。结论:内膜腺上皮和基质细胞HOXA11表达在分泌中晚期变化最显著,而且其表达量呈反相变化,即腺上皮表达量下降,基质细胞表达量增加。提示HOXA11基因与着床期子宫内膜的分化、成熟密切相关;其对内膜腺上皮和基质细胞的调控机制可能不尽相同。     

HOXA10基因在子宫内膜组织中的表达及与不孕的关系

目的　探讨HOXA10基因在正常育龄妇女及不明原因不孕患者子宫内膜中的表达 ,在着床过程中的作用及与不孕的关系。方法　采用原位杂交和逆转录聚合酶链反应 (RT PCR)技术 ,检测 5 2例正常育龄妇女及 38例不明原因不孕患者子宫内膜组织中HOXA10基因mRNA的表达。其中 ,正常育龄妇女子宫内膜周期为 10例增殖早期、10例增殖晚期、9例分泌早期、16例分泌中期、7例分泌晚期 ,不明原因不孕患者子宫内膜周期为增殖早期 7例、增殖晚期 8例、分泌早期 6例、分泌中期 11例、分泌晚期 6例。结果　 (1)HOXA10基因mRNA在正常育龄妇女子宫内膜腺体及间质中均有表达。原位杂交法检测 (以显色区灰度阳性单位表示 )显示 ,分泌中期腺体为 5 6 9± 0 5 7、间质为 7 48± 0 6 7,分泌晚期腺体为 5 99± 0 40、间质为 7 98± 1 0 8,显著高于增殖早期 (腺体 3 0 4± 0 30 ,间质3 2 5± 0 31)、晚期 (腺体 3 35± 0 2 0 ,间质 3 2 0± 0 37)和分泌早期 (腺体 3 0 7± 0 2 6 ,间质 3 18± 0 2 7)(P <0 0 1) ;分泌中、晚期间质表达高于腺体 (P <0 0 1)。RT PCR法检测显示 ,HOXA10基因mRNA表达 ,分泌中期为 (5 7 0± 3 4) %、晚期为 (5 6 2± 2 9) % ,显著高于增殖早期的 (31 8± 2 6 ) %、晚期的(32 2± 2 3) %和     

原位杂交检测同源框基因HoxA11在人月经周期子宫内膜的表达

了解 Hox A1 1基因在人月经周期子宫内膜的表达及变化 ,探索该基因对正常内膜周期的调节功能。应用地高辛标记的 RNA探针进行原位杂交。 Hox A1 1基因主要在内膜间质细胞中表达 ,增殖期与分泌期比较未见明显差异。腺上皮细胞中 Hox A1 1基因的表达量虽较间质细胞少 ,但有明显的周期性变化 ,以增生殖期腺上皮细胞表达量较高 ,分泌早期开始下降 ,到分泌中晚期表达量极低或不表达。Hox A1 1基因在月经周期人子宫内膜中的这种表达时空的变化与孕酮受体 (PR)非常相似 ,提示 Hox A1 1基因的表达与卵巢激素及其受体的调控有着密切的关系 ,Hox A1 1在分泌中期内膜腺上皮中表达下降是内膜分化的重要指标志。     

Hoxa-10在人月经周期分泌中期子宫内膜腺上皮表达下降

目的 :对同源异型框基因 Hoxa-1 0在月经周期各阶段人子宫内膜的表达形式进行观察。方法 :通过原位杂交方法测定了 5例增殖期、9例分泌期的人子宫内膜 Hoxa-1 0的表达。结果 :Hoxa-1 0基因在人子宫内膜均有表达。但在各周期阶段 ,其表达的强度和部位却有所不同。首次发现 Hoxa-1 0在对应于着床期的分泌中期以及分泌晚期子宫内膜腺上皮的表达明显降低或不表达。结论 :Hoxa-1 0在人类着床过程中可能具有重要功能。     

HOXA10的辅因子Meis1、Pbx1在人子宫内膜中的表达

目的:探讨HOXA10的两个辅因子Pbx1和Meis1在人正常子宫内膜中的表达及其变化规律。方法:采用原位杂交进行组织学定位,逆转录聚合酶链反应(RT-PCR)进行半定量,在mRNA水平上检测人正常月经周期子宫内膜中Pbx1和Meis1的表达水平。结果:Meis1在子宫内膜基质细胞和腺上皮细胞均有表达,其中腺上皮细胞的表达高于基质细胞;增生期Meis1仅有弱表达,分泌期显著增加(P<0.05),以分泌中期表达最强(P<0.05)。在整个月经周期中并未检测到Pbx1的表达。结论:Meis1作为HOXA10的辅因子,在人正常子宫内膜中规律性的表达,可能对HOXA10介导的胚胎着床发挥着重要作用。     

左旋18-甲基炔诺酮和米非司酮对分泌中期人子宫内膜HOXA11基因表达的影响

目的:通过口服孕激素类(左旋18-甲基炔诺酮,Levenorgestrel,LNG)和抗孕激素类(米非司酮,RU486)避孕药,研究孕激素对分泌中期人子宫内膜HOXA11基因表达的影响。方法:30名自愿者,于正常月经周期排卵后d7,刮取内膜组织做自身对照;实验周期在排卵前或排卵后2d,口服小剂量LNG(18例)或RU486(12例),同样在排卵后d7刮取内膜组织。用原位杂交方法检测服药子宫内膜HOXA11基因表达的变化。结果:对照组(分泌中期)内膜,间质细胞普遍表达HOXA11mRNA,而腺上皮HOXA11的表达则呈弱阳性或阴性。口服LNG后,子宫内膜形态变化不明显;内膜腺上皮HOXA11表达量进一步下降或无明显变化(即用药前后均表达阴性);间质细胞HOXA11表达增强。服用RU486后,内膜发育明显延迟,腺上皮HOXA11表达强度普遍大于对照组,而间质细胞的表达变化无明显规律。结论:孕激素对内膜腺上皮HOXA11基因的表达起负调控作用;正常分泌中期,内膜腺上皮HOXA11表达减弱或消失是内膜分化成熟的标志。     

HOXA-11基因与不孕及妊娠失败关系的研究

目的 :研究HOXA 11基因与不孕及妊娠失败的关系。方法 :选择 4 1例不明原因不孕症患者及 30例难免流产患者 ,同时选择 2 8例正常未孕者及 18例正常早期妊娠者分别作为对照组 ,留取子宫内膜或蜕膜组织。子宫内膜标本通过组织学检查分为增生期或分泌期。难免流产及正常妊娠组均妊娠 6～ 9周。采用原位杂交方法检测所有子宫内膜或蜕膜组织中的HOXA 11基因mRNA的表达。结果 :整个月经周期子宫内膜腺上皮细胞及间质细胞中均有HOXA 11基因mRNA表达 ,并因月经周期不同而有所波动。HOXA 11基因mRNA在分泌中晚期子宫内膜间质细胞中阳性表达率为 10 0 % ,在增生期子宫内膜间质细胞中的阳性表达率为 6 3.6 % ,差异有显著性 (P <0 .0 5 ) ;在不明原因不孕症患者中HOXA 11基因失去了它在正常子宫内膜表达的周期性变化 ,而且基因表达缺失明显。HOX A 11基因在正常增生期及分泌期子宫内膜腺上皮细胞或间质细胞中的阳性表达率分别为90 .9%、82 .4 %和 6 3.6 %、10 0 % ,在不孕症增生期及分泌期子宫内膜两种细胞的阳性表达率分别为 38.9%、4 7.8%和 2 2 .2 %、39.1% ,两组分别比较 ,差异均有显著性 (P <0 .0 5 ) ;HOXA 11基因在正常早期妊娠蜕膜细胞中持续表达 ,其阳性表达率均为 10 0 % ,在难免流产蜕膜细胞中的阳性表达     

表皮生长因子受体在子宫内膜、早孕蜕膜及滋养细胞中的表达

目的了解表皮生长因子受体（ＥＧＦＲ）在子宫内膜细胞分化及发育中的作用。方法采用免疫组织化学及逆转录聚合酶链反应（ＲＴＰＣＲ）技术测定５８例正常子宫内膜（３２例增生期,２６例分泌期）与２６例早孕蜕膜。结果ＥＧＦＲ存在于正常子宫内膜、早孕蜕膜腺体及间质细胞的细胞膜、核膜及胞浆内,分布均匀。ＥＧＦＲ还位于早孕蜕膜腺体及间质细胞核内。正常子宫内膜ＥＧＦＲ的表达,腺体部分高于间质部分（Ｐ＜０．０５）,ＥＧＦＲ在增生期及分泌期子宫内膜腺体的表达差异无显著性（Ｐ＞０．０５）,但ＥＧＦＲ在早孕蜕膜组织中的表达明显高于增生期及分泌期（Ｐ＜０．０５）。ＥＧＦＲ在间质细胞的表达,早孕蜕膜高于分泌期内膜,而增生期内膜则表达最低（Ｐ＜０．０５）。ＥＧＦＲｍＲＮＡ的表达从弱到强依次为增生期、分泌期、蜕膜、滋养细胞,增生期与分泌期比较,差异无显著性（Ｐ＞０．０５）,早孕蜕膜较分泌期及增生期明显增加（Ｐ＜０．０５）,滋养细胞ＥＧＦＲｍＲＮＡ的表达明显高于增生期、分泌期及早孕蜕膜（Ｐ＜０．０５）。结论ＥＧＦＲ存在于各期子宫内膜中,其表达在正常月经周期中无明显变化,但在早孕蜕膜、滋养细胞中的表达明显高于增生期及分泌期内膜     

月经周期中子宫内膜内皮素及其受体的研究

评价内皮素在内膜生殖生理中的作用,方法利用免疫组织化学和分子原位杂交交结合计算机辅助图象分析技术,检测２３例因良性疾病而行子宫切除术的子宫内膜ＥＴ肽、ＥＴ受体及其两种亚型（ＥＴａＲ、ＥＴｂＲ）的ｍＲＮＡ表达定位、周期性变化。结果在整个月经周期中子宫内膜ＥＴ肽和ＥＴｍＲＮＡ表达有明显的周期性变化,分泌期表达强于增殖期,分泌晚期腺上皮表达最高。基质表达弱于腺上皮,并无明显的周期性变化。ＥＴａＲ和ＥＴｂ     

Apoptosis and Ki-67 expression in adenomyotic lesions and in the corresponding eutopic endometrium.

OBJECTIVE: To examine biologic and proliferative properties of adenomyotic lesions and to determine whether adenomyotic lesions originate in the basal layer of the eutopic endometrium. METHODS: We examined eutopic and ectopic endometria from 23 patients with adenomyosis. To obtain evidence for the induction of programmed cell death, apoptotic cells were identified using a modified terminal deoxynucleotidyltransferase-biotin nick end-labeling method. To evaluate cell death repressor activity, bcl-2 gene expression was examined using immunohistochemical staining. As a proliferative marker, Ki-67 expression was also examined immunohistochemically. RESULTS: In the eutopic endometrium, apoptosis was most frequently observed in epithelial cells during mid- to late secretory phases, although it was rarely found during early proliferative through early secretory phases (P<.01). In contrast, bcl-2 gene expression inversely correlated with the appearance of apoptosis. A similar tendency was observed in stromal cells. In the ectopic endometrium of adenomyosis, endometrial dating revealed that secretory change was rare, even in the secretory phase, and that induction of apoptotic cells as well as bcl-2 gene expression showed no cyclic change. In stromal cells of the ectopic endometrium, apoptosis was more frequent than was seen in the eutopic endometrium, in all menstrual phases (P<.05). Ki-67 was constantly expressed in the glandular epithelium of the ectopic endometrium, irrespective of the menstrual phases, whereas in the secretory phase it was less expressed in the eutopic endometrium of functional and basal layers (P<.01). CONCLUSION: The induction of apoptosis seems to be regulated by hormonal changes in the eutopic endometrium and has an inverse correlation with bcl-2 gene expression. The ectopic endometrium in adenomyosis is rarely influenced by hormonal change and has different biologic and proliferative properties than events observed in the eutopic endometrium findings, which strongly suggest that the adenomyotic lesion does not originate in the basal endometrium.     

Ultrastructural evidence of stromal/epithelial interactions in the human endometrial cycle

We found ultrastructural evidence of interactions between glandular epithelium and superficial stromal cells of the human endometrium during phases of the menstrual cycle. Four significant changes were observed in the transition from early proliferative (days 5 to 9) to early secretory (days 15 to 19) phases. These changes included: (1) an increase in the number and size of lamina densa disruptions, (2) an increase in the number and size of gap junctions, (3) an increase in the number and complexity of epithelial cell projections that extended through the lamina densa, and (4) an increase in close contacts between stromal and epithelial cells. The complex epithelial cell projections that extended through the lamina densa were in close proximity to stromal cells. These interactions were seen primarily in the early secretory phase. After that time (days 20 to 28) the interactions were less frequent. These morphologic results reveal complex physical interactions between epithelial and stromal cells of the adult endometrium. The interactions reach maximal development during the preimplantation phase of the endometrial cycle.     

Differential expression of interleukins (IL) IL-13 and IL-15 throughout the menstrual cycle in endometrium of normal fertile women and women with recurrent spontaneous abortion

Interleukins (IL)-13 and IL-15 are novel cytokines and key regulators of immune/inflammatory related responses that are critical in the outcome of various normal biological and associated abnormalities of the endometrium. The present study determined the temporal and spatial expression of IL-13 and IL-15 mRNA and protein in endometrium of normal fertile women throughout the menstrual cycle, and examined whether profiles of their expression differ from endometrium of women with unexplained recurrent spontaneous abortion (RSA). Using quantitative RT-PCR, ELISA and immunohistochemistry we found that IL-13 and IL-15 mRNA and protein are expressed in control endometrium throughout the menstrual cycle and in RSA (cycle days 21-23) with peak expression detected immediately after and prior to onset of menses, and two distinct periods corresponding to late proliferative and the early-mid secretory phases, respectively. The ratio of IL-13:IL-15 expression revealed a predominance in IL-13 expression during late proliferative/early secretory phase and IL-15 during the mid secretory phase. Compared to control endometrium, endometrium of women with RSA expresses elevated levels of IL-13 and IL-15, with IL-13:IL-15 ratio favoring IL-13. The immunoreactive IL-13 and IL-15 were localized primarily in endometrial luminal epithelial cells with an increased intensity in glandular epithelial and stromal cells in RSA. In conclusion, the results indicate that endometrium of normal fertile women expresses IL-13 and IL-15, with a distinct profile during the menstrual cycle and elevated expression in women with RSA. Although the biological significance of IL-13 and IL-15 in human endometrium and their elevated expression in RSA await investigation, these cytokines with distinct biological functions may regulate endometrial inflammatory/immune responses, tissue repair and receptivity for embryo implantation.     

MMP-26和TIMP-4在人正常月经周期中的表达

目的:观测MMP-26、TIMP-4和ER-α在正常月经周期不同时相子宫内膜组织中的表达及其调节机制。方法:正常子宫内膜组织(n=76)按月经周期时相分为增生早期、增生中期、增生晚期、分泌早期、分泌中期、分泌晚期和月经期。所有标本应用免疫组化和RT-PCR检测MMP-26、TIMP-4和ER-α。结果:MMP-26和TIMP-4蛋白主要定位于子宫内膜腺上皮细胞。增生晚期和分泌早期子宫内膜MMP-26表达高于增生早、中期和分泌中、晚期及月经期子宫内膜。TIMP-4与MMP-26同步表达,月经周期中TIMP-4在增生晚期和分泌早期表达最强。MMP-26mRNA/TIMP-4mRNA表达于增生期开始增强,分泌早期达到高峰,而后下降,至月经期达到最低点。MMP-26启动区-130~-116位点和TIMP-4启动区-930~-916位点间序列与ERE高度同源。结论:人类子宫内膜腺上皮中存在MMP-26和TIMP-4蛋白表达,并呈周期性变化,其表达与子宫内膜功能相关。MMP-26和TIMP-4启动区相关区段与ERE同源,提示ER-α参与MMP-26和TIMP-4基因调节。     

Xanthine oxidase in eutopic and ectopic endometrium in endometriosis and adenomyosis

OBJECTIVE: To investigate the expression of xanthine oxidase in eutopic and ectopic endometrium in endometriosis and adenomyosis. DESIGN: Immunohistochemical identification of xanthine oxidase in endometrial tissues by using polyclonal antibody. SETTING: University hospital. PATIENT(S): Thirty-four women with endometriosis, 34 women with adenomyosis, and 44 fertile control women. INTERVENTION(S): Biopsy samples were obtained from the endometrium throughout the menstrual cycle. MAIN OUTCOME MEASURE(S): Semiquantitative immunostaining (evaluation nomogram) score of endometrial cells. RESULT(S): The level of xanthine oxidase expression in the glandular epithelium of control varied according to menstrual phase, but no such variation in expression was seen in endometriosis. Variation in xanthine oxidase expression was observed during the menstrual cycle in patients with adenomyosis; this variation differed completely from that in controls. Xanthine oxidase expression was found in ectopic endometrial tissue in all cases. The mean evaluation nomogram levels in the glandular epithelium in adenomyosis tissue were as high as those in the early secretory phase in the eutopic endometrium. CONCLUSION(S): Aberrant expression of xanthine oxidase in eutopic and ectopic endometrium appears to play a pathologic role in endometriosis and adenomyosis.     

Expression of Fox head protein 1 in human eutopic endometrium and endometriosis

The objective of this study is to examine the localization and expression of FOXP1 in human endometrium during the menstrual cycle and in endometriotic lesions and endometrial adenocarcinoma. FOXP1 protein expression was analyzed by immunohistochemistry and Western blot. FOXP1 expression was significantly different between glandular epithelial and stromal nuclei and cytoplasm in both endometrial functionalis and basalis. FOXP1 immunostaining was significantly reduced in the early secretory stage in comparison to the mid proliferative stage in the functionalis and the early proliferative stage in the basalis. FOXP1 expression was found in endometriotic lesions but not in endometrial adenocarcinoma. Multiple protein bands of FOXP1 were identified, and their presence varied considerably among patients. Protein expression levels were significantly higher in the mid and late secretory stages in comparison to early proliferative and early secretory stages. FOXP1 protein is present in human endometrium with evidence of cycle stage-dependent changes in expression.     

Immunohistochemical analysis of proliferative activity and steroid receptor expression in peritoneal and ovarian endometriosis

Objective: To assess the proliferative activity of eutopic and ectopic endometrium throughout the menstrual cycle and its correlation to steroid receptor content.Design: The immunohistochemical use of Ki 67 was applied to investigate the proliferation index. A recently advanced stereographic computer technology was used to investigate steroid receptors.Setting: University hospital department of gynecology.Patient(s): Biopsies of eutopic endometrium, black and red peritoneal endometriotic lesions, and ovarian endometriomas were taken from infertile patients and classified according to the phase of the cycle.Result(s): In normal endometrium, the glandular proliferation index was highest during the proliferative phase and was statistically significantly reduced during the secretory phase. No proliferative activity was observed in the late secretory phase. No statistically significant differences were found between ectopic endometrium and eutopic endometrium except during the late secretory phase, when proliferative activity was still present in endometriotic tissue. The stromal proliferation index was similar in red lesions, ovarian endometriomas, and eutopic endometrium during the secretory phase. In normal endometrium, the highest concentrations of estrogen receptors (ERs) and P receptors (PRs) occurred in the epithelial and stromal cells during the late proliferative phase of the menstrual cycle. Estrogen receptor and PR content declined throughout the secretory phase. In ectopic endometrium, PR persisted in the glandular epithelium during the late secretory phase. Estrogen receptors persisted in the glandular epithelium and stroma of red peritoneal lesions and ovarian endometriomas during the late secretory phase.Conclusion(s): The high proliferative activity and the persistence of ERs and PRs in the stroma of red lesions and ovarian endometriomas emphasize the primordial role of the stroma in the development of endometriosis and suggest different mechanisms of proliferation control from those observed in eutopic endometrium.     

Expression of aminopeptidase N in human endometrium and regulation of its activity by estrogen

Objective: To determine whether aminopeptidase N (APN) regulates the cycle-dependent bioavailability of interleukin-8 (IL-8) in the endometrium.

Design: Prospective study.

Setting: University medical center.

Patient(s): Women without endometrial pathology from the proliferative (n = 25) or secretory (n = 18) phase of the menstrual cycle.

Intervention(s): We first immunolocalized APN in the endometrium using an anti-APN antibody. We then determined the regulation of APN kinetic activity by sex steroids in endometrial stromal cell cultures.

Main Outcome Measure(s): Expression of APN in human endometrium throughout the menstrual cycle. Regulation of APN activity by estradiol and progesterone in cultured endometrial stromal cells.

Result(s): Immunohistochemistry of endometrial sections revealed staining of endometrial stroma throughout the menstrual cycle. There was no detectable staining in glandular cells. The expression of APN as detected by immunohistochemistry was significantly lower in the early proliferative phase. In cultured cells, estradiol inhibited APN activity in a concentration-dependent manner. Progesterone did not have a significant effect.

Conclusion(s): Stromal localization of APN in endometrium may explain the epithelial rather than stromal presence of IL-8 in vivo. Decreased expression of APN may increase IL-8 bioavailability thus contributing to angiogenesis and polymorphonuclear leukocyte chemotaxis in early proliferative phase.