Expression of interleukin-8 receptors in endometriosis

BACKGROUND: Although the etiology of endometriosis is not well understood, chemokines and their receptors are believed to play a role in its pathogenesis. Therefore, we aimed to investigate the expression and localization of interleukin-8 (IL-8) receptors CXCR1 and CXCR2 in eutopic and ectopic endometrial tissues of women with endometriosis, and in endometrium of women without endometriosis. METHODS: Ectopic (n = 27) and homologous eutopic endometrium (n = 25) from women with endometriosis and endometrium from women without endometriosis (n = 27) were used for immunohistochemical analysis of CXCR1 and CXCR2. RESULTS: In normal endometrium, epithelial CXCR1 and CXCR2 immunostaining intensities were similar in the proliferative and secretory phase. Stromal CXCR1 expression was less then epithelial expression and did not show cyclical difference. No stromal CXCR2 expression was observed. In eutopic endometrium of women with endometriosis compared to endometrium of women without endometriosis, there was a significant increase in both proliferative and secretory phases for epithelial CXCR2 expression, and in proliferative phase for CXCR1 expression (P < 0.05). Both receptor immunoreactivities were significantly increased in the epithelial cells of ectopic endometrial tissues compared to that of normal endometrium (P < 0.05). CONCLUSIONS: These findings suggest that IL-8 and its receptors may be involved in the pathogenesis of endometriosis.     

Immunohistochemical characterization of proliferation, oestrogen receptor and progesterone receptor expression in endometriosis: comparison of eutopic and ectopic endometrium with normal cycling endometrium

Recent studies examining oestrogen and progesterone receptorstatus and the proliferative activity of endo-metriotic lesionshave produced conflicting reports. This study aimed to clarifythe receptor status and proliferative activity of eutopic andectopic endometrium from women with endometriosis and endometriumfrom normal women. Progesterone and oestrogen receptor expressionand proliferative activity were studied in eutopic and ectopicendometrium from 30 women with endometriosis and in endometriumfrom 30 normal cycling women using microwave-pretreated paraffin-embeddedsections stained with an avidin-biotin peroxidase technique.Progesterone and oestrogen receptor expression in the controlendo-metrium did not differ from that of eutopic endometriumfrom women with endometriosis. Oestrogen receptor expressionin ectopic endometrium increased from the proliferative to thelate secretory phase. Epithelial progesterone receptor expressiondecreased during the cycle. Oestrogen receptor expression inboth epithelium and stroma of ectopic endometrium was significantlyhigher than in eutopic endometrium throughout the cycle. Incontrast, stromal progesterone receptor expression tended tobe reduced in ectopic endometrium compared with eutopic tissue.Epithelial progesterone receptor expression was increased inectopic endometrium but only in the late secretory phase. Althoughproliferative activity in the epithelium of control and eutopicendometrium was reduced from the proliferative to the late secretoryphase, stromal activity did not vary. The proliferative activityin ectopic endometrium remained low and constant throughoutthe cycle. In the proliferative and early secretory phases,the proliferative activity of eutopic endometrium was increasedcompared with ectopic endometrium, but in the late secretoryphase, levels were comparable. These findings challenge previousreports which have suggested that oestrogen receptors are reducedin ectopic tissue. This may have clinical implications for thedevelopment of novel treatments for endometriosis.     

Phenotypic and functional studies of leukocytes in human endometrium and endometriosis

The aetiology of endometriosis, a common and disabling disorder, is presently unknown, although immune dysfunction could allow ectopic endometrial fragments to survive outside the uterine cavity. These studies investigate the relationship between leukocyte populations, steroid hormone receptor expression, proliferative activity, bcl-2 expression and apoptosis in eutopic and ectopic endometrium from women with endometriosis or adenomyosis at different phases of the menstrual cycle. Significantly increased oestrogen receptor expression, bcl-2 expression and numbers of CD8+ leukocytes were found in ectopic compared with eutopic endometrium in endometriosis, and CD56+ endometrial granulated lymphocytes (eGLs) were significantly reduced in ectopic endometrium. Apoptotic cells were rarely found in control and subject endometria. In contrast with endometriosis, adenomyotic lesions showed identical steroid hormone receptor expression, proliferative activity, bcl-2 expression and leukocyte subpopulations to eutopic endometrium, indicating different aetiologies for these disorders. The unusual CD56+ CD16- eGLs present in large numbers in late secretory phase eutopic endometrium were highly purified (>98%) by immunomagnetic separation. Except for a negligible cytotoxic activity of eGLs from early proliferative samples, cytotoxic activity of eGLs from non-pregnant endometrium during the menstrual cycle was comparable with those in peripheral blood, predominantly CD56+ CD16+ natural killer cells. eGLs from non-pregnant endometrium and early pregnancy showed a variable proliferative response to 5 and 100 U/ml interleukin-2 over 48-h and 120-h time courses. eGLs are evidently functionally important in the eutopic endometrium. Their absence in endometriotic lesions together with increased CD+8 T-cell numbers and increased oestrogen receptor and bcl-2 expression may have significant effects on the development and progression of endometriosis.     

Apoptosis and bcl-2 expression in normal human endometrium, endometriosis and adenomyosis

Apoptosis has been implicated in the pathogenesis of several diseases and is partly regulated by bcl-2, which blocks the apoptotic pathway and promotes cell survival. Apoptosis and bcl-2 expression were examined in paired eutopic and ectopic endometrium from women with endometriosis (n = 30 samples) or adenomyosis (n = 15 samples) and compared with control endometrium (n = 30 samples). Apoptotic cells were detected using the dUTP nick-end labelling (TUNEL) assay for DNA fragmentation; bcl-2 expression was demonstrated with a streptavidin- biotin peroxidase immunohistochemical technique. Apoptotic cells were rare in eutopic, ectopic and control endometrium; there were no significant differences between subject groups nor between eutopic and ectopic endometrium. Stromal bcl-2 expression increased in the late secretory phase in control and eutopic endometrium in endometriosis; double labelling studies revealed that most stromal bcl-2+ cells were leukocytes. Stromal bcl-2 expression in endometriotic foci was significantly increased compared with the paired eutopic endometrium, did not vary with menstrual cycle and included a significant population of non-leukocytic bcl-2+ stromal cells. In contrast, stromal bcl-2 expression in adenomyosis remained at low levels and did not show significant cyclical variation. Glandular epithelial bcl-2 expression also varied with menstrual cycle phase and peaked in the proliferative phase; in contrast, surface epithelial bcl-2 expression increased in the late secretory phase. Elevated stromal bcl-2 expression in ovarian endometriotic lesions could have implications for the growth and survival of ectopic endometrial tissue at these sites.      

Cytokine network of eutopic and ectopic endometrium in women with adenomyosis

PROBLEM: Recent studies showed the impairment of local cytokine balance in women with external endometriosis, but similar findings concerning direct production of cytokines by immunocompetent cells of women with adenomyosis are absent. In this context, investigation of the cytokine synthesis by mononuclear cells (MNCs) infiltrating eutopic and ectopic endometrium is of special interest. METHOD OF STUDY: Concentration of interferon-gamma (IFNgamma), interferon-alpha (IFNalpha), tumour necrosis factor-alpha (TNFalpha), interleukin- 1beta (IL-1beta) and epidermal growth factor (EGF) in supernatants (SNs) of 24-hr cultures of MNCs obtained from eutopic and ectopic endometrium of women with adenomyosis was determined by enzyme-linked immunosorbent assay. RESULTS: The levels of IFNgamma, IFNalpha, TNFalpha, IL-1beta and EGF in SNs of eutopic endometrial MNCs of women with adenomyosis were significantly increased and the content of IL-8 in SNs was reduced compared with that of the control figures. Ectopic MNCs of women with adenomyosis produced higher levels of IFNgamma, IFNalpha and TNFalpha than the MNCs of normal endometrium. The production of IL-1beta, IL-8 and EGF by ectopic endometrial MNCs was significantly reduced. CONCLUSION: The results obtained indicate a significant role of local cytokine production impairment in the development of adenomyosis.     

galectin-3在子宫内膜异位症患者在位内膜中的表达及意义

目的：探讨galectin-3在子宫内膜异位症（内异症）患者在位子宫内膜中的表达及意义.方法：分别采用免疫组化和实时定量逆转录-聚合酶链反应（Real-time RT-PCR）检测内异症患者在位内膜及正常对照中galectin-3蛋白及mRNA的表达,比较其表达差异.结果：galectin-3蛋白定位于子宫内膜的腔上皮细胞、腺上皮细胞和间质细胞.与正常对照相比,galectin-3 mRNA及蛋白在内异症在位内膜中的表达显著下降（P＜0.05）.其表达在分泌期高于增生期.结论：内异症患者在位子宫内膜galectin-3低表达可能与其子宫内膜容受性的改变有关,从而导致了内异症不孕.     

GnRHⅡ蛋白在子宫内膜异位症患者中的表达及意义

目的：检测GnRHⅡ蛋白在子宫内膜异位症患者异位子宫内膜、在位子宫内膜和正常子宫内膜中的表达情况,同时分析其表达是否与子宫内膜月经周期有关。方法：采用免疫组织化学SP法检测GnRHⅡ蛋白在异位内膜、在位内膜及正常子宫内膜组织中的表达情况,并分析和比较其表达是否有差异。结果：GnRHⅡ蛋白在子宫内膜异位症患者异位、在位子宫内膜及正常子宫内膜中均有表达,阳性表达定位于子宫内膜腺体及间质细胞的细胞质;GnRHⅡ蛋白在异位内膜、在位内膜及对照组正常内膜的表达依次增强,两两比较差异有统计学意义（P＜0.05）;GnRHⅡ蛋白在正常子宫内膜分泌期表达强于增生期（P＜0.05）,且以分泌早中期最强,显著强于增生期和分泌晚期（P＜0.01）,而异位组或在位组的分泌期与增生期比较,差异无统计学意义（P＞0.05）。结论：GnRHⅡ蛋白在子宫内膜异位症的发病中以及在人类月经生理方面可能起重要作用。     

ORIGINAL ARTICLE: The Characterization of the Subpopulation of Suppressive B7H4+ Macrophages and the Subpopulation of CD25+ CD4+ and FOXP3+ Regulatory T‐cells in Decidua during the Secretory Cycle Phase,Arias Stella Reaction,and Spontaneous Abortion – A Preliminary Report

Problem The presence of immunosuppressive cells within the endometrium and decidua is crucial for establishing maternal immune tolerance against fetal antigens. We decided to evaluate the subpopulations of Treg cells and B7H4 macrophages in eutopic endometrium typified by Arias Stella reaction during the development of Fallopian tube pregnancy as well as in decidua at the time of spontaneous abortion (SA), and to compare these findings to those observed in the endometrium during the secretory cycle phase of healthy women. Method of study The decidual tissue samples evaluated in our study were obtained from 26 women who underwent curettage as a result of the following circumstances: five of the women because of a laparoscopic procedure necessitated by Fallopian tube pregnancy, and 11 of them because of SA. The control group consisted of 10 patients on whom curettage was preformed as an additional procedure during laparoscopic myomectomy. The presence of regulatory T‐cells and B7H4‐positive macrophages in the samples was analysed by fluorescence‐activated cell sorter (FACScan). Results Both the percentages of FOXP3⁺ cells in the subpopulation of CD25⁺ CD4⁺ T lymphocytes and the percentage of B7H4‐positive cells in the macrophage subpopulation found in the deciduae of patients suffering SA were higher than those found in eutopic endometrium with Arias Stella reaction. No such differences in the percentages of these cells were observed when the tissue samples from patients with SA were compared with those from the control group. The percentage of B7H4‐positive macrophages, however, was found to be significantly lower in endometrium with Arias Stella reaction in comparison to that observed in secretory endometrium. Conclusion The alterations in both the Treg cell and suppressive B7H4⁺ macrophage subpopulations would seem to be related to the suppression of maternal immune cells in the endometrium at the beginning of decidualization.     

基质金属蛋白酶2及其抑制因子2在子宫腺肌病中的表达及意义

目的 探讨基质金属蛋白酶2(MMP-2)及其抑制因子2(TIMP-2)在子宫腺肌病(AM)在位内膜与异位内膜的表达及意义.方法 选取广州医学院附属广州市第一人民医院2006年2月至2007年9月因AM行全子宫切除术的42例病例(取其在位及异位内膜分为AM在位内膜组和AM异位内膜组)和同期因子宫肌瘤在该院行全子宫切除术的32例病例(取其内膜作为正常子宫内膜组),应用半定量反转录聚合酶链反应(RT-PCR)检测AM在位内膜、异位内膜及正常子宫内膜中MMP-2、TIMP-2mRNA的表达.结果 (1)MMP-2 mRNA在AM异位内膜组中相对表达量(1.43±0.32)高于AM在位内膜组和正常子宫内膜组(1.22±0.35,0.97±0.37,P＜0.05).TIMP-2 mRNA在AM异位内膜组和AM在位内膜组中均低表达,其相对表达量分别低于正常子宫内膜组(0.96±0.31,1.13±0.30,1.29±0.19,P＜0.05).(2)MMP-2 mRNA、TIMP-2 mRNA在子宫腺肌病在位内膜组、正常子宫内膜组的表达有周期性,分泌期均高于增生期(P＜0.05);而在AM异位内膜组中,两者分泌期与增生期则差异无统计学意义.(3)MMP-2/TIMP-2的比值在AM异位内膜组、AM在位内膜组、正常子宫内膜组中依次递减,其中AM异位内膜组与后两组差异有统计学意义(均P＜0.05).结论 AM异位和在位内膜组织中MMP-2高表达,TIMP-2低表达,使MMP-2/TIMP-2平衡失调,异位内膜组织侵袭降解细胞外基质的能力增强.     

血管内皮生长因子在子宫内膜异位症发病中的作用

目的探讨血管内皮生长因子(vascularendothelialgrowthfactor,VEGF)在子宫内膜异位症(endometriosis,EM)发病中的作用。方法应用免疫组织化学方法并结合图像分析技术。结果正常子宫内膜和EM在位内膜腺上皮细胞的VEGF随月经周期呈现规律性变化,分泌期腺上皮VEGF蛋白表达量显著高于增殖期(P<0.05)。在增殖期,EM在位子宫内膜腺上皮VEGF的表达与正常子宫内膜相比无明显差别,但在分泌期,EM在位子宫内膜腺上皮细胞中VEGF的表达强度明显高于正常子宫内膜(P<0.01)。EM在位内膜腺上皮的VEGF含量显著高于同组卵巢子宫内膜异位囊肿的异位腺上皮(P<0.01)。结论表明VEGF的表达异常与EM的发病有关。     

In vitro apoptosis effects of GnRHII on endometrial stromal cells from patients with endometriosis

Purpose: To study the effect of gonadotropin-releasing hormone II (GnRHII) on the cell apoptosis of ectopic, eutopic and normal endometrial stromal cells cultured in vitro from endometriosis patients, and to provide theoretical basis for exploring new treatments for endometriosis (EMs). Methods: Ectopic, eutopic and normal endometrial stromal cells were isolated, cultured and identified in vitro, then treated with different concentrations of GnRHII (0, 10^-10 M, 10^-8 M and 10^-6 M). Cell apoptosis was detected by Hoechst staining and flow cytometry. Results: GnRHII increased apoptosis in ectopic, eutopic and normal stromal cells in a dosage-dependent manner (P<0.05), and apoptosis of ectopic stroma cells was significantly higher than that of eutopic and normal cells (P<0.05); apoptosis in eutopic and normal cells had no different (P>0.05). Conclusion: GnRHII can significantly induce apoptosis in ectopic, eutopic and normal endometrial stromal cells from patients with endometriosis, especially to the ectopic.     

STAT3和SOCS3与子宫内膜异位症的相关性研究

目的: 探讨信号转导和转录活化因子3(STAT3)和细胞因子信号转导抑制因子3(SOCS3)的表达与子宫内膜异位症的相关性。方法: 采用Western blotting方法检测30例子宫内膜异位症患者异位(异位组)、在位子宫内膜(在位组)和25例正常子宫内膜(对照组)中总STAT3、磷酸化STAT3(p-STAT3)和SOCS3蛋白的表达情况。结果: 总STAT3蛋白在3组内膜中表达无差异,而其活化水平(p-STAT3/STAT3)在异位组最高,SOCS3蛋白表达在异位组最低,两者与在位和对照组相比较差异有统计学意义(P<0.05),相关性分析显示内异症患者中STAT3活化水平与SOCS3蛋白表达呈负相关(r=-0.499, P<0.01)。结论: 在子宫内膜异位症患者的异位子宫内膜组织中存在STAT3过度活化和SOCS3蛋白表达下降,两者呈负相关。     

Immunohistochemical analysis of oestrogen and progesterone receptors of eutopic and ectopic endometrium in the rabbit model of endometriosis: the effect of pregnancy.

Oestrogen receptors (ER) and progesterone receptors (PR) were measured in the rabbit model of endometriosis in eutopic and ectopic endometrium of pregnant animals (n = 7) and controls (n = 7). Immunostaining of cryostat sections of ectopic and eutopic endometrium was performed using monoclonal antibodies against ER and PR. Levels of ER and PR were 'evaluated' in a semi-quantitative manner using a modified histoscore. The ER and PR content in stromal and glandular cells was not different in eutopic and ectopic endometrium in either pregnant or non-pregnant control animals. There was a significant difference between the PR content of the glandular epithelium of pregnant animals and controls for both eutopic (P less than 0.02) and ectopic (P less than 0.001) endometrium. The disappearance of glandular PR and the persistence of stromal PR suggest that the function of the glandular endometrium is mediated by the paracrine and autocrine action of stromal cells. The decidual cells are likely to produce substances that may be of importance in embryo implantation and early pregnancy.     

HLA-G蛋白在子宫腺肌病中的表达及其作用机制

目的:探讨HLA-G蛋白在子宫腺肌病中的作用机制。方法:采用免疫组织化学染色方法与Western blot方法检测31例子宫腺肌病患者在位与异位内膜组织HLA-G蛋白表达,18例子宫肌瘤患者正常子宫内膜作为对照。结果:子宫腺肌病患者在位与异位内膜组织腺上皮、间质细胞均有HLA-G蛋白表达,而子宫肌瘤患者(对照组)正常内膜中未检测到HLA-G蛋白;子宫腺肌病患者在位与异位内膜组织HLA-G蛋白表达无显著性差异(P0.05);子宫腺肌病患者在位内膜分泌期和增生期HLA-G蛋白表达无显著性差异(P0.05)。结论:子宫腺肌病患者在位与异位内膜组织HLA-G蛋白异常表达,可能与其发病有关。     

Decreased apoptosis and sensitivity to macrophage mediated cytolysis of endometrial cells in endometriosis

Ectopic dissemination of endometrial cells and their subsequent implantation are the mechanisms involved in the development of endometriosis. While the process of dissemination appears to be a phenomenon common to all women, it is unknown what facilitates or prevents ectopic implantation of misplaced endometrial cells. Prior studies by our group and others suggest that cell-mediated immunity in patients with endometriosis is decreased. The present studies evaluated (i) peripheral blood monocyte (PBM) and peritoneal macrophage (PM) mediated cytolysis of autologous eutopic and ectopic endometrial cells and (ii) programmed cell death (apoptosis) in the eutopic and ectopic endometrium. PBM-mediated cytolysis was (mean+/-SD) 23.1+/-13% for the eutopic and 7.8+/-% for the ectopic endometrium (P < 0.004), while the corresponding percentages for PM-mediated cytolysis were 5.4+/-7 and 0.3+/-1 respectively (P < 0.04). This indicates that PBM are much more effective than PM in inducing cytolysis of both eutopic and ectopic endometrium and that ectopic endometrial cells are significantly more resistant to both PBM- and PM-mediated cytolysis. The apoptosis was significantly decreased in the eutopic endometrium of women with endometriosis as compared to fertile controls (0.375+/-0.17 versus 1.57+/-0.3, P < 0.0001). Furthermore, in matched samples apoptosis was significantly lower in the ectopic (0.149+/-0.075) than eutopic (0.375+/-0.17) endometrium (P < 0.001). We conclude from these studies that the decrease in the capacity of monocytes to mediate cytolysis of the misplaced endometrial cells in the peritoneal locations and an increased resistance of these cells to apoptosis are fundamental to the aetiology and/or pathophysiology of endometriosis.     

Endometrial cells from women with endometriosis have increased adhesion and proliferative capacity in response to extracellular matrix components: towards a mechanistic model for endometriosis progression

BACKGROUND: Endometriosis, classified as the presence of endometrial cells in ectopic sites, is a debilitating disease causing pain and infertility in approximately 10% of women of reproductive age. It is associated with the aberrant expression of extracellular matrix (ECM) components and their receptors, integrins. METHODS: We analysed the expression of integrins in stromal cells derived from peritoneal, ovarian and deeply infiltrating endometriotic lesions and from endometrium from women with and without endometriosis in vitro, using quantitative immunocytochemistry. The adhesive and proliferative capacity of each of the cell types in response to ECM components was assessed by in vitro assays of cell attachment and DNA synthesis. RESULTS: We demonstrate that eutopic and ectopic endometrial stromal cells from women with endometriosis exhibit an aberrant integrin profile in vitro compared with stromal cells derived from healthy controls. In addition, the former display increased adhesion and proliferative capacity in response to specific ECM components. CONCLUSIONS: We propose that the increased adhesive and proliferative potential of cells from endometriotic lesions may be a key feature in the pathogenesis of endometriosis. Furthermore, the elevated responsiveness of eutopic cells from women with endometriosis may contribute to the predisposition of some women to the disease.     

组蛋白去乙酰化酶1在子宫内膜异位症中的表达及意义

目的 研究组蛋白去乙酰化酶1(HDAC1)在子宫内膜异位症患者在位及异位内膜中的表达,探讨其在子宫内膜异位症发生、发展中的作用. 方法 应用免疫组织化学和免疫印迹法检测20例子宫内膜异位症患者在位内膜和异位内膜组织(研究组)中及20例子宫肌瘤患者的子宫内膜组织(对照组)HDAC1的表达情况. 结果 HDAC1阳性着色主要分布于子宫内膜上皮细胞和间质细胞的细胞核,在位内膜中HDAC1的表达强度明显高于对照组子宫内膜(P<0.01).免疫印迹检测提示,子宫内膜异位症在位内膜和异位内膜组织中HDAC1蛋白的相对表达量分别为2.67±0.69和2.55±1.36,显著高于对照组子宫内膜1.63±0.93(P<0.01,P<0.05);而在位内膜组与异位内膜组之间无统计学差异(P＞0.05). 结论 HDAC1在子宫内膜异位症在位和异位内膜组织中的高表达,可能在子宫内膜异位症的发生、发展中起重要作用.     

Distribution of cyclooxygenase-2 in eutopic and ectopic endometrium in endometriosis and adenomyosis

The objective of this study was to determine the distribution of cyclooxygenase-2 (COX-2) in eutopic and ectopic endometria in endometriosis and adenomyosis. The subjects were 35 patients with endometriosis diagnosed by laparoscopy, 33 patients with histologically confirmed adenomyosis and 50 female controls with normal fecundity. Expression of COX-2 was immunohistochemically investigated in tissues from eutopic endometrium and myometrium and ectopic endometrium of the wall of ovarian chocolate cysts using polyclonal antibody. Surface epithelial cells, endometrial glandular epithelial cells or stromal cells were assessed. Cells were semi-quantitatively assessed on a scale of 1 to 5 using a nomogram created from positive cell count and the degree of staining. COX-2 expression in surface and glandular epithelia of the control group varied markedly during the menstrual cycle. It was lowest in the early proliferative phase and gradually increased thereafter. It remained high throughout the secretory phase. However, in patients with endometriosis, expression of COX-2 in glandular epithelium was higher than that in the control group, though it varied throughout the menstrual cycle. On the other hand, there was no variation in expression of COX-2 in the adenomyosis group during the menstrual cycle, and it was lower than that in the endometriosis group in all phases. Pronounced COX-2 expression was observed in glandular cells from ectopic endometrial tissue of ovarian chocolate cyst walls in all cases regardless of the menstrual phase. In summary, increased COX-2 expression in eutopic and ectopic endometria was believed to be strongly correlated with pathological abnormalities in these disorders.