血管内皮生长因子及其受体在月经周期子宫内膜中的表达

目的　检测血管内皮生长因子 (VEGF)及其受体fms样酪氨酸激酶 (flt 1)、含插入区的激酶受体 (KDR)在正常子宫内膜中的表达 ,探讨其在子宫内膜血管生成及内膜生长分化中的作用。方法　采用免疫组织化学及原位杂交方法 ,对 5 0例妇女正常月经周期子宫内膜中VEGF、flt 1、KDR蛋白质及mRNA进行检测 ,用蛋白印迹法分析子宫内膜中VEGF亚型 ,用内皮细胞标志物Ⅷ因子标记内膜微血管密度。结果　VEGF、flt 1、KDR蛋白质及mRNA主要分布在正常子宫内膜血管内皮细胞及腺上皮细胞 ,VEGF、flt 1在分泌中期、月经期的表达较强 ,KDR在增生中期即达高峰 ,三者在增生早期的表达均最低。分泌期血管壁面积及血管腔面积大于增殖期 (P <0 0 5 ) ,血管密度无明显周期性改变 (P >0 0 5 )。正常子宫内膜组织 4种VEGF蛋白亚型中 ,以VEGF12 1、VEGF16 5为主 ,其免疫条带积分吸光度明显高于VEGF189、VEGF2 0 6 (P <0 0 5 )。结论　VEGF、flt 1、KDR蛋白质及mRNA在正常子宫内膜中的表达呈明显周期依赖性 ,分泌期表达增强提示其可能参与胚胎着床 ,月经期表达增加可能与此期子宫内膜脱落缺氧有关     

Effect of mifepristone on estrogen and progesterone receptors in human endometrial and endometriotic cells in vitro

OBJECTIVE: To investigate the expressions of estrogen (E) receptor and progesterone (P) receptor in human eutopic and ectopic endometrium and the effect of mifepristone (RU486) on them. DESIGN: Prospective study. SETTING: University hospital. PATIENT(S): Twenty-two patients with ovarian endometriosis and 13 patients with uterine leiomyoma were recruited. INTERVENTION(S): Samples of ovarian endometrioma cyst tissue and endometrium were obtained from the 22 patients. A sample of endometrium was obtained from the 13 patients. MAIN OUTCOME MEASURE(S): Expressions of E and P receptors were determined using immunocytochemical method. RESULT(S): P receptor expression in endometrial epithelial cells with endometriosis was significantly higher than that without endometriosis in the early secretory phase. Estrogen receptor and epithelial P receptor expressions in endometriotic cells were significantly lower than those of endometrial cells during the proliferative phase, similar with the latter during the early secretory phase and significantly higher during the late secretory phase. RU486 down-regulated the expressions of E and P receptors in both the eutopic and the ectopic endometrial cells, and in some cases this down-regulating effect was more apparent when the concentration of RU486 was higher. CONCLUSION(S): Different steroid receptor expressions indicate different hormonal regulation between endometriotic and endometrial cells. The down-regulating effect on E and P receptors may be one of the therapeutic mechanisms of RU486 on endometriosis.     

Endometrial osteopontin, a ligand of beta3-integrin, is maximally expressed around the time of the "implantation window"

OBJECTIVE: To analyze endometrial mRNA and protein expression of osteopontin and its receptor, beta3-integrin, throughout the menstrual cycle. DESIGN: Study by immunohistochemistry, RNase protection assay, and ELISA. SETTING: Academic research unit. PATIENT(S): Forty-five regularly cycling women without endometrial pathology. INTERVENTION(S): Expression of endometrial osteopontin and beta3-integrin mRNA was analyzed by RNase protection assay in endometrium, endometrial epithelial cells, stromal cells, and endometrial leukocytes (CD45) and by immunohistochemistry in frozen sections of endometrium throughout the menstrual cycle. Concentration of osteopontin was studied in uterine secretions by ELISA. MAIN OUTCOME MEASURE(S): mRNA and protein expression of osteopontin and beta3-integrin. RESULT(S): Osteopontin mRNA and protein was weakly expressed in the proliferative phase and maximally expressed in the mid to late secretory phases in endometrium, endometrial epithelial cells, and endometrial leukocytes and in uterine secretions. Beta3-integrin mRNA and protein were expressed in stromal cells throughout the menstrual cycle and were maximally expressed in epithelial cells in the mid to late secretory phases. CONCLUSION(S): Expression of osteopontin and its receptor, beta3-integrin, in human endometrial glands and osteopontin secretion into the uterine cavity around the time of the "implantation window" suggest a role for these proteins in endometrial function and implantation.     

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     

子宫内膜端粒酶活性的研究

目的　探讨月经周期中各期子宫内膜的端粒酶表达及其在子宫内膜周期性变化中的意义。方法　采用端粒重复序列扩增法 (telomericrepeatamplificationprotocol,TRAP)方法检测 6 3例正常子宫内膜和 19例子宫内膜癌组织的端粒酶活性。结果　月经周期中不同期的子宫内膜的端粒酶活性表达不同。增殖期阳性率为90 6 % (2 9/ 32 ) ;分泌期阳性率为 4 7 4 % (9/ 19) ,二者之间差异有显著性 (P =0 0 0 0 6 )。绝经期子宫内膜也有5 0 0 % (6 / 12 )表现为阳性 ,子宫内膜癌阳性率为 89 5 % (17/ 19)。增殖期子宫内膜有 5 3 1% (17/ 32 )端粒酶活性为强阳性而分泌期和绝经期子宫内膜的端粒酶活性则没有强阳性。结论　月经周期中子宫内膜细胞的周期性变化 ,端粒酶可能起着重要的调控作用     

Localization of immunoglobulin-containing cells in human endometrium in the first trimester of pregnancy and throughout the menstrual cycle

The distribution of immunoglobulins in normal human endometrium throughout the menstrual cycle and in early pregnancy has been studied with an immunoperoxidase technique. In first-trimester decidua, IgG was detected within many cells of differing morphology and size. Large IgG-containing cells were often binucleate and were believed to be decidual cells. Examination of serial sections showed no kappa or lambda light-chain restriction, suggesting absorption of the immunoglobulin content. Medium-sized, irregular, IgG-containing cells were macrophages. An additional substantial population of small hyperchromatic IgG-containing cells were prominent around arterioles and adjacent to endometrial glands. From examination of adjacent sections stained with phloxine tartrazine, it was concluded that these represented endometrial granulocytes. Labelling for light chains again suggested absorption of the immunoglobulin content. In contrast, in non-pregnant endometrium immunoglobulin-containing stromal cells were uncommon, although IgG and IgA were detected in gland epithelium and secretions and in the stromal interstitium particularly in the secretory phase. These results support the notion that human endometrium lacks a classical secretory immune system and highlight the requirement for correlation between studies of cell surface markers, morphology and cell surface receptors.     

Fluctuation of 6Ckine expression in human endometrium during the menstrual cycle

OBJECTIVE: To clarify the expression of 6Ckine, a potential chemoattractant for endometrial natural killer (NK) cells, in the human endometrium. DESIGN: Experimental study. SETTING: Department of obstetrics and gynecology at a medical university. PATIENT(S): Fifty-seven fertile women 25 to 52 years of age who had regular menstrual cycles and normal endometrium and were undergoing hysterectomy. INTERVENTION(S): Endometrium was obtained from operative samples. MAIN OUTCOME MEASURE(S): Tissue was immunostained to determine the localization of 6Ckine in the endometrium throughout the menstrual cycle. The number of NK cells was counted in 10 nonoverlapping stromal areas. The concentration of 6Ckine in homogenized endometrium was determined by enzyme-linked immunosorbent assay. RESULT(S): Endometrial surface, glandular epithelial cells, and perivascular stromal cells were immunoreactive for 6Ckine throughout the menstrual cycle with some fluctuation. In addition, some T cells, NK cells, and macrophages in the stroma were immunoreactive for 6Ckine. The 6Ckine concentration was low in the proliferative phase but elevated in the secretory phase. It showed a moderate positive correlation with the number of endometrial NK cells. CONCLUSION(S): 6Ckine may be a potential chemoattractant for endometrial NK cells.     

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     

Activation antigens during the proliferative and secretory phases of endometrium and early-pregnancy decidua

BACKGROUND: Clarifying the normal distribution of activation antigens will contribute to database construction studies of monoclonal-antibody-based therapies in endometrial disorders. METHODS: In this study, endometrial tissue samples obtained during proliferative and secretory phases and decidual samples of early pregnancies were immunostained by the monoclonal antibodies anti-CD26, anti-CD30, anti-CD70, anti-CD71, and anti-CD98 using the indirect immunoperoxidase method. RESULTS: CD26 is expressed on the glandular epithelium in the endometrium and decidua. Endothelial CD26 is expressed less in the decidua when compared to the endometrium. CD30 is strongly expressed by decidual cells. It is only weakly expressed on endometrial and decidual vessels. Glandular and endothelial CD70 expression is mainly seen in the proliferative phase of the menstrual cycle. Glandular CD71 expression is less in the decidua when compared to the endometrium. Its expression on stromal cells is more in the secretory phase of the menstrual cycle and in early pregnancy deciduae. It is expressed on endometrial vessels but not on decidual vessels. Glandular CD98 is expressed more in the decidua when compared to the endometrium. This antigen exists on endometrial lymphocytes. It is strongly expressed on the endothelium in the endometrium and decidua. CONCLUSION: It seems that CD26 and CD70 are not involved in the functions of endometrial and decidual stromal cells. CD30 and CD71 are thought to be involved in decidualization. Absence of activation antigens other than CD98 on lymphocytes indicated an antigenic profile for large granular lymphocytes that is different from regular lymphocytes.     

Temporal and morphologic characteristics of pinopod expression across the secretory phase of the endometrial cycle in normally cycling women with proven fertility

OBJECTIVE: To assess the temporal and morphologic characteristics of pinopod expression on the surface of endometrium across the secretory phase, in LH-timed endometrial samples in normal, healthy women. DESIGN: Prospective, randomized study. SETTING: Academic teaching hospital. PATIENT(S): Sixty-eight healthy volunteers with proven fertility. INTERVENTION(S): Urinary LH-timed endometrial and blood sampling was performed on each subject on a randomly selected day of the secretory phase. MAIN OUTCOME MEASURE(S): Histologic dating, assessment of pinopods using scanning electron microscopy, and comparison with serum P levels. RESULT(S): Eighty-six endometrial tissue samples obtained from 68 subjects were evaluated under scanning electron microscopy. Pinopods were first observed on luteal day 5, corresponding with the onset of the midluteal phase increase in serum P levels. Pinopods persisted for the entire duration of the secretory phase, but their morphology changed as the cycle advanced. CONCLUSION(S): The present findings demonstrate that pinopods are a characteristic feature of the mid to late secretory phase endometrial epithelium, exhibit cycle-dependent changes in morphology, and are most prominent during the putative implantation interval.     

血管内皮生长因子受体KDRmRNA和flt-1mRNA与子宫内膜异位症相关性研究

目的探讨血管内皮生长因子受体(VEGFR)中的fms样酪氨酸激酶受体(flt-1)和胎肝激酶受体(KDR)在子宫内膜异位症(内异症)患者异位及在位子宫内膜组织中的表达及其意义.方法采用RT-PCR技术,从核酸水平检测53例内异症患者(44例卵巢子宫内膜异位囊肿,30例内异症在位子宫内膜组织)中的VEGF受体KDRmRNA、flt-1mRNA的阳性表达率及相对表达强度,以40例正常子宫内膜组织作为对照.结果与对照组比较,研究组KDRmRNA和flt-1mRNA的阳性表达率高于对照组,差异有显著性(P＜0.05);研究组在位内膜增殖期与分泌期的阳性表达率、相对表达强度高于对照组子宫内膜组织,差异有显著性(P＜0.05);不同月经周期在位内膜组织KDRmRNA和flt-1mRNA的表达、两组增殖期KDRmRNA相对表达强度高于分泌期flt-1mRNA,而flt-1mRNA分泌期相对表达强度高于KDRmRNA.结论VEGF受体KDRmRNA和flt-1mRNA在内异症中有明显表达并呈周期依赖性.VEGF受体在内异症血管生成中的作用,可能与内异症远处转移、侵袭、种植相关.     

子宫内膜异位症淋巴细胞亚群分布研究

目的:本研究检测子宫内膜异位症(EMs)患者在位和异位内膜组织中CD45RO,CD4和CD8阳性的T淋巴细胞数量表达,探讨免疫细胞在EMs中的作用,为免疫治疗提供依据。方法:采用CD45RO,CD4和CD8单克隆抗体,应用免疫组织化学法标记EMs患者的在位和异位子宫内膜组织中CD45RO,CD4和CD8阳性细胞,观察其分布特点并计数其密度,并与正常妇女子宫内膜对照。结果:EMs组和正常对照组的增殖期与分泌期子宫内膜组织中均存在CD45RO,CD4和CD8阳性的淋巴细胞,EMs组在位和异位内膜中CD45RO,CD8阳性细胞数量均高于对照组,差异有统计学意义(P<0.05),且在异位内膜中的表达较在位内膜升高,差异具有统计学意义(P<0.05);在位和异位内膜中CD4阳性细胞较对照组略有升高,差异无统计学意义(P>0.05);在位和异位内膜中CD4/CD8的比值均较对照组降低,差异有统计学意义(P<0.05);在位和异位内膜之间比较CD4阳性细胞、CD4/CD8的比值无明显变化,差异无统计学意义(P>0.05)。在位内膜的CD45RO,CD4和CD8阳性细胞表现为由增殖期到分泌期增加的趋势,但差异无统计学意义(P>0.05);异位内膜中CD45RO,CD4及CD8阳性细胞无明显增殖期与分泌期的改变(P>0.05);对照组中CD45RO及CD4阳性细胞从增殖期到分泌期有增多的趋势,但差异无统计学意义(P>0.05)。EMs组异位和在位内膜Ⅰ～Ⅱ期病例和Ⅲ～Ⅳ期病例之间CD45RO,CD4和CD8阳性细胞未发现明显变化,差异无统计学意义(P>0.05)。结论:EMs患者在位和异位内膜中T淋巴细胞亚群分布的改变为EMs的发生提供条件,并对EMs的发生、发展和维持起重要作用。     

Analysis of normal human endometrial stroma cells--identification and cell kinetics of stromal cells in the human endometrium using the combined technic of immunohistochemistry and 3H-thymidine autoradiography

Combined Method of immunohistochemistry using monoclonal antibody to leucocyte-common antigen and autoradiography after incorporation of radiothymidine in vitro were carried out in human endometrium to identify the cell type of various stromal cells and to clarify their cell kinetics in a sexual cycle. It became evident that stromal fibroblasts incorporated radiothymidine during the proliferative phase only, while leucocytes, reacted with the monoclonal antibody, also did during the late secretory phase. The cells of endometrial blood vessels incorporated radiothymidine during the whole cycle. Endometrial granulocytes, carrying leucocytecommon antigen, incorporated radiothymidine during the late secretory phase.     

Metallothionein and RCAS1 expression in comparison to immunological cells activity in endometriosis, endometrial adenocarcinoma and endometrium according to menstrual cycle changes

OBJECTIVE: Endometrium is a specialized organ in which phenomena controlling the level of cell proliferation and apoptosis are marked. The aim of our study was to determine the presence of proteins involved in apoptosis and proliferation: RCAS1, MT and the number of CD56-positive cells and their activity to elucidate their possible role in the development of adenocarcinoma and endometriosis. MATERIALS AND METHODS: MT, RCAS1, CD56-positivity and CD69 expression were assessed in 55 tissue samples by Western blot and immunohistochemistry methods. RESULTS: We found that endometrium during secretory menstrual cycle phase is characterized by significantly higher RCAS1 and higher MT expression than in proliferative phase. The number of CD56-positive cells and the CD69 antigen expression was significantly increased. Endometrial adenocarcinoma was characterized by significantly increased RCAS1 expression, while MT expression was comparable to the level found in the secretory phase. The number of CD56-positive cells was significantly decreased and their activity was comparable to the level found in the secretory phase. Endometriosis was accompanied by significantly lower RCAS1 and MT expressions, with lower number of CD-56 positive cells and lower expression of CD69 antigen in comparison to the secretory phase. CONCLUSIONS: The ability of endometrium to determine cytotoxic activity (RCAS1 expression changes) and high protection against DNA damage (MT expression) with concomitant changes in the number of immune cells and their activity, observed in normal endometrium during the menstrual cycle phases seems to be fundamental for pathological features of endometrial adenocarcinoma and endometriosis.     

Antigen-presenting cells in human endometrium during the menstrual cycle compared to early pregnancy

OBJECTIVE: Human endometrium and early pregnancy decidua harbor a considerable and diverse population of antigen-presenting cells (APC). Changes in the number and distribution of macrophages and dendritic cells (DC) could point to a possible role of these immunocompetent cells in implantation and success of early pregnancy. METHODS: Uterine tissue was obtained from 22 women undergoing hysterectomy for bleeding disorders or dysmenorrhea and from 11 women undergoing legal abortion. Tissue was investigated with antibodies against CD14, CD68, CD83, DC-SIGN, Ki-67, and human leukocyte antigen (HLA)-DR using single and double immunohistochemical staining techniques. RESULTS: The number of CD14(+) cells was stable during all phases of the menstrual cycle and early pregnancy. In comparison to nonpregnant endometrium, DC-SIGN(+) cells showed a higher proliferation rate and were found associated in clusters with CD56(+) natural killer (NK) cells in early pregnancy. In the late secretory phase of the menstrual cycle, numbers of CD83(+) (P <.01) cells were significantly higher than in other endometrial phases and early pregnancy. HLA-DR(+) expression was significantly increased in early pregnancy but remained unchanged throughout the menstrual cycle. CONCLUSION: The presence of DC-SIGN(+) cells during the menstrual cycle and their proliferation in early pregnancy suggests an important role of these cells with regard to the balance between defense against pathogens and tolerance of the fetal allograft. Whether the increase of CD83(+) mature DC and CD68(+) macrophages in the late secretory phase is caused by hormonal stimuli and/or is due to changes of the cytokine/chemokine micromilieu remains to be investigated.     

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.     

The differential expression of oestrogen receptors, progesterone receptors, Bcl-2 and Ki67 in endometrial polyps

Objective To obtain a greater understanding of the pathogenesis of endometrial polyps and to gain insight into which factors play a pivotal role in their growth.
Design Retrospective analysis of archived paraffin-embedded specimens.
Setting St James's University Hospital.
Sample Thirty secretory phase endometrial samples, 10 secretory phase endometrial polyps, 8 proliferative phase endometrial samples and 10 proliferative phase endometrial polyps.
Methods Immunohistochemistry was used to characterise the expression of oestrogen and progesterone receptors, Bcl-2 and Ki67 in cycling endometrium and phase-matched endometrial polyps. Patterns of expression were compared between the polyps and the endometrium.
Main outcome measure The expression of oestrogen receptors, progesterone receptors, Bcl-2 and Ki67.
Results Three significant differences were found between the endometrium and the polyps. Polyps taken from the proliferative phase of the cycle displayed significantly elevated expression of Bcl-2 and weak or no expression of progesterone receptors. Secretory phase polyps displayed an elevated expression of oestrogen receptors.
Conclusion A localised increase in Bcl-2 expression and consequential decline or cessation of apoptosis is an important mechanism underlying the pathogenesis of endometrial polyps. Elevated Bcl-2 expression results in failure of the polyp tissue from undergoing normal cyclical apoptosis during the late secretory phase. This may mean the polyp is not shed along with the rest of the endometrium during menstruation.