interleukin-8 and Endometriosis

Endometriosis is a chronic inflammatory disease associated with an impairment in immune response. Disorders in the peritoneal fluid and ectopic endometrium macrophage populations and their secretory products create a specific microenvironment inducing the development of the disease. The important factors involved in inflammation associated with endometriosis are chemokines, especially interleukin (IL)-8. For this reason, the current study briefly reviews the role of IL-8 in the pathogenesis of endometriosis.. A systematic review was done on all published studies that compared IL-8 expression and concentration in patients with and without endometriosis to evaluate their potential as biomarkers for the disease.. IL-8 induces chemotaxis of neutrophils and other immune cells; also, it is a potent angiogenic agent. Most researchers pointed to the increased peritoneal and serum IL-8 levels and showed correlation with the severity of the disease, size and number of the active lesions. IL-8 takes part in all processes during the development of the disease: adhesion, invasion, and implantation of ectopic tissue. Additionally, the chemokine plays a role in growth and maintenance of ectopic endometrial tissue directly affecting endometrial cell proliferation. IL-8 might also protect ectopic cells against death by apoptosis.. It may act as an autocrine growth factor in the endometrium and promotes the vicious circle of endometrial cell attachment and, in consequence, may lead to a transformation from acute to chronic inflammation stage.

Topics: Biomarkers; Cellular Microenvironment; Endometriosis; Female; Humans; Interleukin-8; Peritoneum

Can we use chemokines as biomarkers to diagnose patients with endometriosis in clinical practice?. Some chemokines, especially CXCL8 (IL-8), CCL-2 (MCP-1) and CCL5 (RANTES), have the potential to work as biomarkers to identify patients with endometriosis but their accuracy could be improved by combination with other non-inflammatory markers in a panel of biomarkers.. The need for a good marker to diagnose endometriosis has increased in recent years and research in this field has intensified. Chemokines have been reported to be associated with endometriosis in several studies over the last 20 years. Many of these studies measured one or more chemokines in peritoneal fluid (PF) and peripheral blood (PB) or through endometrial biopsies in patients with and without endometriosis.. A systematic review was done on all published studies that compared chemokine concentrations in patients with and without endometriosis to evaluate their potential as biomarkers for the disease.. Using MEDLINE database from December 1993 to August 2013 and the MeSH terms 'Endometriosis' and 'Chemokines', we identified relevant studies to include in the present review, which was based on the PRISMA statement. Studies that measured at least one chemokine in patients with endometriosis and matching controls in PB, PF or endometrial samples were included. We did not include samples from ectopic lesions. All review articles as well as studies with animals and those not written in English were excluded from this systematic review. The studies were assessed using a modified version of the Quality Assessment of Diagnostic Accuracy Studies criteria. Two authors independently assessed studies for inclusion and risk of bias, and extracted data.. After inclusion and exclusion criteria, 62 studies were selected to be included in this systematic review. A total of 27 different chemokines or their receptors were evaluated in the reviewed studies. The most studied chemokines (including their receptors) were CXCL8 (51.6%), CCL2 (38.7%) and CCL5 (19.3%) (% of studies). CXCL8 (IL-8) appears to have the best results among all the other chemokines as a marker for endometriosis.. Some studies included have low power due to small sample size and study designs vary in the assessment criteria for the markers, the state of the patients (e.g. phase of the cycle and stage of disease) and the nature of the controls.. Our findings could guide future research in this field to select the chemokines with the best potential, and to stimulate better-designed studies to determine whether they can become a useful diagnostic tool in clinical practice.. There was no funding to support this systematic review. The authors have no competing interest to declare.

Topics: Ascitic Fluid; Biomarkers; Chemokine CCL2; Chemokine CCL5; Chemokines; Endometriosis; Endometrium; Female; Humans; Inflammation; Interleukin-8

Apoptosis is a physiologic process that eradicates undesired cells without inducing an inflammatory reaction. It is an important regulator of eutopic endometrial function and evidence suggests that apoptosis aids in maintaining cellular homeostasis during the menstrual cycle by eliminating aging cells from the functional layer of the uterine endometrium. Endometriosis, which is characterized by the growth of endometrial tissue outside the uterus, could result from increased cellular proliferation or decreased apoptosis in response to appropriate stimuli. Eutopic endometrium from women with endometriosis has several differences compared with normal endometrium of women without endometriosis. These differences may contribute to the survival of regurgitated endometrial cells into the peritoneal cavity and thus to the development of endometriosis. In this article, we will summarize recent literature concerning apoptosis-related genes such as Bcl-2 and Fas, outline the molecular basis of apoptosis and review the literature focused on the alterations in regulation of apoptosis in eutopic and ectopic endometrium from women with endometriosis.

Topics: Apoptosis; Caspases; Endometriosis; Endometrium; Epithelium; Fas Ligand Protein; fas Receptor; Female; Gene Expression Regulation; Humans; Interleukin-8; Peritoneum; Proto-Oncogene Proteins c-bcl-2; Receptors, Tumor Necrosis Factor

To review the malignant potential of endometriosis based on epidemiologic, histopathologic, and molecular data.. Literature review.. The pathogenesis of endometriosis remains unclear. The histopathologic development of endometriosis has undergone long-term investigation. Studies have confirmed histologic transition from benign endometriosis to ovarian malignancy, including malignant transformation of extraovarian endometriosis. The prevalence of endometriosis in patients with epithelial ovarian cancer, especially in endometrioid and clear cell types, has been confirmed to be higher than in the general population. Ovarian cancers and adjacent endometriotic lesions have shown common genetic alterations, such as PTEN, p53, and bcl gene mutations, suggesting a possible malignant genetic transition spectrum. Furthermore, endometriosis has been associated with a chronic inflammatory state leading to cytokine release. These cytokines act in a complex system in which they induce or repress their own synthesis and can cause unregulated mitotic division, growth and differentiation, and migration or apoptosis similar to malignant mechanisms.. The malignant potential of endometriosis holds serious implications for management, such as the need for earlier and more meticulous surgical intervention for complete disease treatment.

Topics: Cell Transformation, Neoplastic; Endometriosis; Female; Gene Expression Regulation, Neoplastic; Genomic Instability; Humans; Incidence; Intercellular Signaling Peptides and Proteins; Interleukin-1; Interleukin-8; Mutation; Ovarian Neoplasms; Prevalence; Transforming Growth Factor beta; Treatment Outcome; Tumor Necrosis Factor-alpha

Endometriosis, defined by the presence of viable endometrial tissue outside the uterine cavity, is among the most common gynecologic disorders affecting women of reproductive age. Endometriosis is associated with an inflammatory peritoneal environment, where multiple cytokines and growth factors are found at elevated levels. Interleukin-8 (IL-8) is a cytokine that induces chemotaxis of neutrophils and is a potent angiogenic agent. In addition, IL-8 was recently found to stimulate proliferation of various cells. We have observed that IL-8 is elevated in the peritoneal fluid of women with endometriosis and the levels correlate with the severity of the disease. We hypothesized that IL-8 may play a role in the growth and maintenance of ectopic endometrial tissue not only by chemoattracting and stimulating leukocytes to secrete growth factors and cytokines, but also by directly affecting endometrial cell proliferation. We found that IL-8 mRNA and protein levels in the endometrium were significantly higher during early proliferative and late secretory phases than during the mid-cycle. IL-8 receptors A and B are also expressed in the endometrium mostly localized in the stroma. Interestingly, IL-8 receptor expression is higher in the eutopic endometrium of women with endometriosis compared to the endometrium of women without endometriosis. Endometrial cells in culture proliferate significantly when treated with IL-8, which is inhibited by anti-IL-8 neutralizing antibody. More convincingly, IL-8 antisense oligonucleotide treatment decreases IL-8 production by endometrial cells as well as cell proliferation when compared to non-sense oligonucleotide treatment. The addition of IL-8 reverses the inhibitory effect of IL-8 antisense oligonucleotides on cell proliferation. These findings suggest that IL-8 may act as an autocrine growth factor in the endometrium. We have also studied the effect of endometrial cell adhesion on IL-8 expression and observed that IL-8 stimulates the adhesion of endometrial cells to fibronectin. Treatment of the cells with anti-IL-8 neutralizing antibody inhibited partially the cell adhesion. Thus, IL-8 may also be relevant for stimulating the attachment of endometrial implants in the pathogenesis of endometriosis. In addition, adherence of endometrial cells induced further IL-8 expression by an integrin-dependent mechanism. In summary, IL-8 may act as an autocrine growth factor in the endometrium and may also play a role in the pathogenesis

Topics: Cell Adhesion; Cell Division; Endometriosis; Endometrium; Female; Humans; Interleukin-8

Chemokines are increasingly recognized as important regulators of uterine function.. The following is a review of uterine chemokines, especially monocyte chemotactic protein (MCP)-1, interleukin (IL)-8, and regulated-upon-activation normal-T-cell-expressed and -secreted (RANTES) protein, in reproductive physiology and pathology.. It is increasingly clear that IL-8, MCP-1, RANTES and their receptors are produced by endometrial, myometrial, and trophoblast cell types in a timed and co-ordinated manner. In addition to the regulation of leukocyte migration and function, uterine chemokines also display specific roles in endometrial angiogenesis, apoptosis, proliferation, and differentiation. IL-8, MCP-1 and RANTES are regulated by local growth factors and cytokines such as tumor necrosis factor-alpha (TNF-alpha), interferon-gamma, and IL-1. IL-8 takes part in cervical ripening and parturition. IL-8, MCP-1 and RANTES are also found at high levels in the peritoneal fluid of women with endometriosis.. Co-ordination of chemokine-chemokine receptor interactions plays an important role in the menstrual cycle and successful pregnancy. Moreover, unbalanced chemokine expression contributes to pathologic conditions typified by uncontrolled cellular proliferation, migration and invasion.

Topics: Cervical Ripening; Chemokine CCL2; Chemokine CCL5; Chemokines; Endometriosis; Endometrium; Female; Humans; Interleukin-8; Menstruation; Myometrium; Parturition; Pregnancy; Receptors, Chemokine; Reproduction; Signal Transduction; Uterus

Topics: Angiogenesis Inducing Agents; Endometriosis; Endothelial Growth Factors; Estrogens; Female; Fibroblast Growth Factor 2; Humans; Interleukin-8; Lymphokines; Thymidine Phosphorylase; Vascular Endothelial Growth Factor A; Vascular Endothelial Growth Factors

Among angiogenic factors, VEGF secreted from activated macrophages under the influence of ovarian steroids, IL-8 expressed in endometrial stromal cells, and basic FGF expressed in endometriotic tissue and PD-ECGF expressed in lining epithelial cells independently of the sex steroidal milieu might contribute to the characteristic advancement of angiogenic lesions in endometriosis in individual manners. Copyrightz1999S.KargerAG,Basel

Topics: Adult; Endometrial Neoplasms; Endometriosis; Endometrium; Endothelial Growth Factors; Female; Fibroblast Growth Factor 2; Gene Expression Regulation, Neoplastic; Humans; Interleukin-8; Lymphokines; Neovascularization, Pathologic; Thymidine Phosphorylase; Vascular Endothelial Growth Factor A; Vascular Endothelial Growth Factors

Chemokines have been associated with endometriosis. Our study was aimed at evaluating the levels of six chemokines--CXCL8 (IL-8), CXCL12 (SDF-1), CCL2 (MCP-1), CCL5 (RANTES), CCL19 (MIP-3β), and CCL21 (6-Ckine)--in the peritoneal fluid (PF) of patients with and controls without endometriosis by multiplexed cytokine assay. In this retrospective case-control study conducted at the Charité University Hospital, patients (n = 36) and controls (n = 27) were enrolled. The patients were separated into groups according to stage of the disease: I-II (n = 21), III-IV (n = 1 5), and according to clinical findings: peritoneal endometriosis (PE; n = 7), deep-infiltrating endometriosis (DIE) affecting the retrocervical area (n = 13) or the bowel/rectovaginal site (n = 14). The subjects were also separated according to the cycle phase: follicular (n = 14) or luteal (n = 8) and the previous use (n = 25) or not (n = 38) of hormones. PF was collected from all subjects (n = 63) consecutively during laparoscopy. The concentration of chemokines in the PF was assessed using Luminex(®) x-MAP(®) technology. Sensitivity and specificity were calculated. A model of multiple logistic regressions estimated the odds of endometriosis for each combination of the chemokines detected. We observed significantly higher concentrations of IL-8 (p < 0.001), MCP-1 (p = 0.014), and MIP-3β (p = 0.022) in the PF of women with endometriosis than in the controls. A joint evaluation revealed that elevated levels of the three chemokines had a positive endometriosis prediction value of 89.1%. The combined assessment of MCP-1, MIP-3β, and IL-8 concentration in PF improved the likelihood of identifying patients with endometriosis. Future studies should investigate this panel in peripheral blood samples.

Topics: Adult; Ascitic Fluid; Chemokine CCL19; Chemokine CCL2; Endometriosis; Female; Follicular Phase; Humans; Interleukin-8; Luteal Phase; Retrospective Studies

Cytokines, and specifically interleukin 6 (IL-6) and interleukin 8 (IL-8), have been associated with the pathogenesis of endometriosis. We studied serum concentrations of IL-6 and IL-8 in patients with deep infiltrating endometriosis (DIE) or ovarian endometriomas (OE), but no other forms of associated endometriosis disease type. We carried out a case-control study including 19 patients with OE alone (OE group), 14 patients with DIE alone (DIE group) and 24 healthy patients without endometriosis (C group). Serum concentrations of IL-6 and IL-8 were measured in the three groups of patients. Serum levels of both IL-6 and IL-8 were significantly higher in the OE group. A high positive correlation was found between serum IL-6 and IL-8 levels in the OE group but not in the DIE and C groups. Serum IL-8 alone achieved the highest predictive value of the presence of OE (adjusted OR: 1.44; sensitivity: 78.2%; specificity: 76.2%). The combination of IL-6 and IL-8 levels did not significantly improve the discrimination between subjects with OE and those with DIE over that of IL-8. OE but not DIE are associated with increased serum levels of IL-6 and IL-8, and thus these may become useful tools for discriminating OE alone from DIE.

Topics: Adolescent; Adult; Case-Control Studies; Endometriosis; Female; Humans; Interleukin-6; Interleukin-8; Ovarian Diseases

Endometriosis is a common gynaecological disorder characterized by the presence of endometrial tissue outside the uterine cavity. This disease is associated with pelvic inflammation and displays some features of autoimmune disorder. Human neutrophil peptides 1, 2, and 3 (HNP 1-3) belonging to α-defensin family play a crucial role in innate immunity against infections and may exert immunoregulatory effects. They may play a role in various inflammatory reactions; however, their role in endometriosis has not been studied. Therefore, the aim of the present study was to evaluate HNP 1-3 in the peritoneal fluid of 67 patients with endometriosis and 16 healthy control women in relation to peritoneal leukocyte subpopulations (neutrophils, T cells, and macrophages) and inflammatory cytokines (IL-6 and IL-8). HNP 1-3, IL-6 and IL-8 were evaluated in the peritoneal fluid by specific enzyme-linked immunosorbent assays (ELISA), and peritoneal leukocyte subpopulations were evaluated by flow cytometry. We found that the levels of HNP 1-3 were significantly increased in the peritoneal fluid of endometriosis patients, compared with control women, and correlated with severity of the disease. Endometriosis was also associated with increased concentrations of peritoneal neutrophils. In endometriosis the levels of HNP 1-3 strongly correlated with concentrations of neutrophils, T cells and IL-8. HNP 1-3 levels were not associated with peritoneal IL-6 or macrophages. These data suggest that HNP 1-3 and neutrophils might play a role in immunopathogenesis of endometriosis and may be worth evaluating as targets for anti-endometriosis therapy.

Topics: Adult; alpha-Defensins; Ascitic Fluid; Biomarkers; Case-Control Studies; Defensins; Endometriosis; Enzyme-Linked Immunosorbent Assay; Female; Humans; Immunity, Innate; Interleukin-8; Neutrophils; T-Lymphocytes

To assess the angiogenic activity of peritoneal fluid in women with minimal to mild endometriosis and to investigate the relationship between this activity and the concentration of macrophage-derived angiogenic factors and clinical variables, such as phase of menstrual cycle, type of lesion, and revised American Society for Reproductive Medicine classification.. In vivo bioassay.. Tertiary-care university medical center.. Fifty-two female volunteers with laparoscopic findings indicating minimal to mild endometriosis.. Peritoneal fluid was collected at the start of laparoscopy. A standard amount of peritoneal fluid was applied to a chick embryo chorioallantoic membrane assay.. Angiogenic response was assessed by determining the vascular density index.. 85% of the peritoneal fluid samples induced angiogenesis in the chick embryo chorioallantoic membrane bioassay. Tumor necrosis factor-alpha and total protein were significantly related to the vascular density index, whereas interleukin-1beta, interleukin-8, and clinical variables appeared to not affect the angiogenic response.. The results confirms previous findings of peritoneal fluid angiogenic activity in women with minimal to mild endometriosis and indicate involvement of tumor necrosis factor-alpha.

Topics: Adult; Allantoin; Animals; Ascitic Fluid; Chick Embryo; Chorion; Endometriosis; Female; Humans; Interleukin-1; Interleukin-8; Laparoscopy; Membranes; Neovascularization, Pathologic; Proteins; Regression Analysis; Tumor Necrosis Factor-alpha

To evaluate interleukin (IL)-1β, IL-6, IL-8, and IL-12p70 levels in serum and peritoneal fluid in women related to infertility and pelvic pain.. Eighty-seven women were diagnosed with endometriosis or cases related to infertility. IL-1β, IL-6, IL-8, and IL-12p70 levels in serum and peritoneal fluid were determined by enzyme-linked immunosorbent assay (ELISA). Pain assessment was evaluated by the Visual Analog Scale (VAS) score.. Serum IL-6 and IL-12p70 levels increased in women with endometriosis compared to the control group. Serum and peritoneal IL-8 and IL-12p70 levels correlated with VAS scores in infertile women. A positive correlation was also found between peritoneal IL-1β and IL-6 with VAS score. A significant difference in peritoneal IL-1β levels was associated with menstrual pelvic pain, while peritoneal IL-8 levels were related to dyspareunia, menstrual, and post-menstrual pelvic pain in infertile women.. An association of IL-8 and IL-12p70 levels were related to pain in endometriosis, as well as a relationship between cytokine expression and VAS score. Further studies should be addressed to investigate the precise mechanism of cytokine-related pain in endometriosis.

Topics: Cytokines; Endometriosis; Female; Humans; Infertility, Female; Interleukin-12; Interleukin-6; Interleukin-8; Pelvic Pain

Endometriosis presents a pro-inflammatory microenvironment influenced by cytokines, such as interleukin (IL)-8, which expression may be influenced by genetic polymorphisms. Therefore, we aimed to investigate the role of interleukin (IL)-8 rs4073 polymorphism in endometriosis' development and its related symptoms. A case-control study was conducted with 207 women with endometriosis and 193 healthy controls. Polymorphism was genotyped using a TaqMan validated assay. Associations were evaluated by binary logistic regression, using odds ratios (OR) and 95 % confidence intervals (CI), and P ≤ 0.05 was considered significant. Cases were younger (36 ± 6.8 versus 39 ± 8.4) and had lower body mass index (26.5 ± 5.3 versus 35.7 ± 6.3 Kg/m

Topics: Case-Control Studies; Endometriosis; Female; Genetic Predisposition to Disease; Humans; Interleukin-8; Interleukins; Pelvic Pain; Polymorphism, Genetic

Endometriosis is a common gynecological hurting disorder in which tissue is similar to the tissue that normally lines the inner layer of the uterus. It often causes fertility problems. Unfortunately, effective treatments are limited. Therefore it's important to explore an imperative and easily accessible treatment to alleviate the probable pathologies and preserve fertility in endometriosis. Consequently, we aimed to investigate the effects of metformin, letrozole, and atorvastatin on inflammation and apoptosis in experimentally induced ovarian and peritoneal endometriosis in rat models. In the present study, 35 rats were randomly divided into five groups. Group 1: sham-operated control group. Group 2: untreated endometriosis group. Group 3: given 100 mg/kg/day of oral metformin. Group 4: given 0.1 mg/kg/day of oral letrozole. Group 5: given 2.5 mg/kg/day of oral atorvastatin. At the end of the 28 days, we examined Ki67, Bax and Bcl-2 immunoexpressions in ovarian and peritoneal tissues, and IL-6, IL-8, and TNF-α levels were evaluated from the peritoneal fluid. All medical treatment groups showed a significant decrease in Ki67 expression. A significant increase in Bax expression was also observed in all samples from all medical treatment groups (other than the untreated endometriosis groups). Further, a significant decrease in Bcl-2 expression was found in all medical treatment groups. IL-6, IL-8, and TNF-α levels were significantly lower in all medical treatment groups than in the endometriosis groups. In conclusion; Metformin, letrozole, and atorvastatin showed apoptosis induction and anti-inflammatory effects on both ovarian and peritoneal endometriosis in experimental models.

Topics: Animals; Apoptosis; Atorvastatin; bcl-2-Associated X Protein; Endometriosis; Female; Humans; Inflammation; Interleukin-6; Interleukin-8; Ki-67 Antigen; Letrozole; Metformin; Proto-Oncogene Proteins c-bcl-2; Rats; Tumor Necrosis Factor-alpha

Endometriosis is a gynecological disease that endometrial cells develop outside the uterus. This event happens when the endometrial glands grow outside the endometrium and uterine muscles, especially in the pelvis. Although endometriosis is widespread, the clinical manifestations of the disease are very different, and it is challenging to adapt to the conventional classification system to divide patients into homogeneous groups. Given the importance of endometriosis, a correct, accurate, and timely diagnosis of this disease can significantly prevent its complications. Using health-related software is one of these ways. Enhanced Endometriosis Archiving Software (ENEAS) is a web-based application based on one of the most widely used open-source database management systems (MySQL), allowing the direct link to other open-source software for data management and storage. In the current study, the effect of ENEAS application was considered in patients with endometriosis, and its influence on IL-8 and MCP-1 gene expression was evaluated. For this purpose, 100 women with endometriosis were divided into two groups of 50 patients. The first group (control group) was examined by a gynecologist and received medication and treatment. In the case group, their demographic and clinical information were entered into ENEAS software. To study the expression of the IL-8 gene and MCP-1 gene, after collecting 5 ml of blood samples in tubes containing anticoagulant, RNA extraction was performed by Total RNA Purification Kit (Cat. 17200, 37500, 17250). Then cDNA synthesis was performed for this purpose, and a Bioneer DNA synthesis kit (South Korea) was used. The results showed that the expression level of the IL-8 gene in the case group was significantly reduced compared to the control group (P = 0.035). MCP-1 gene expression was also decreased compared to the control group, but this decrease was not significant. Therefore, those who used this application for treatment had reduced expression of IL-8 and MCP-1 genes. This event indicates that this application has reduced the amount of inflammation caused by endometriosis with proper analysis.

Topics: Endometriosis; Endometrium; Female; Humans; Interleukin-8; RNA; Software

Endometriosis is an inflammatory estrogen-dependent gynecological disease characterized by the presence of endometrial tissue outside the uterine cavity. An important role in the pathogenesis of this disease is played by disorders of the immune system involving chemokines and their receptors, including the CXCL8-CXCR1/ 2 system.. The aim of the study was to assess the concentration of the CXCL8 chemokine and its CXCR1 and CXCR2 receptors in the peritoneal fluid of women with endometriosis.. The study included 32 women aged 21 to 47 years with diagnosed endometriosis and a control group of 8 healthy women aged 21 to 40 years. The material for the research was the peritoneal fluid collected during the laparoscopic procedure. The concentration of chemokines was determined by ELISA tests.. The conducted studies showed that the concentration of the CXCL8 chemokine was significantly higher in the peritoneal fluid of the studied women and depended on the clinical advancement of the disease.. Changes in the concentration of the CXCL8 chemokine in the peritoneal fluid of women with endometriosis may indicate impaired immune response and indicate an inflammatory process within the peritoneal cavity. The demonstrated relationship between the concentration of CXCL8 and the stages of clinical advancement indicates a significant role of this chemokine in the development of the disease.

Topics: Ascitic Fluid; Chemokines; Endometriosis; Female; Humans; Interleukin-8; Receptors, Interleukin-8A; Receptors, Interleukin-8B

to identify novel biomarkers for peritoneal endometriosis in eutopic endometrium thus giving an oportunity for non-invasive diagnosis.. A cross-sectional single-center study SETTING: tertiary care hospital PATIENTS: 49 patients subjected to laparoscopy because of suspected endometriosis, 33 patients out of the group qualified to the study had sufficient endometrial tissue taken and were in their follicular phase of menstrual cycle.. biopsy sampling of eutopic endometrial tissue during diagnostic or diagnostic and terapeutic laparoscopy, questionaires, MAIN OUTCOME MEASURE(S): qRT-PCR to evaluate the mRNA expression of selected candidate marker genes in endometrium: ARO1 (aromatase), CXCL8 (interleukin 8), NGF (nerve growth factor), VEGF-A (vascular endothelial growth factor A), PDGF-A (platelet-derived growth factor A).. mRNA expression of ARO1, CXCL8, VEGF-A and PDGF-A did not differ significantly between women with and without endometriosis. NGF mRNA expression was decreased in women with endometriosis.. Observed preliminary results suggest a possible role of NGF in early diagnosis of peritoneal endometriosis. The role of NGF changes in eutopic endometrium of patients with peritoneal endometriosis needs further evaluation.

Topics: Adult; Aromatase; Biomarkers; Case-Control Studies; Cross-Sectional Studies; Endometriosis; Endometrium; Female; Gene Expression; Humans; Interleukin-8; Nerve Growth Factor; Peritoneal Diseases; Platelet-Derived Growth Factor; RNA, Messenger; Tertiary Care Centers; Vascular Endothelial Growth Factor A

Inflammation and immune responses play crucial roles in the development of endometriosis. Although interleukin-9 (IL-9) has a pro-inflammatory function in chronic inflammatory diseases, its function in endometriosis remains unknown. Here, we aimed to investigate the significance of IL-9 and IL-9-producing lymphocytes in endometriosis.. Specimens were obtained from patients with and without endometriosis. Peritoneal fluid (PF), peripheral blood (PB), and ovarian endometrioma (OE) tissues were analyzed for the proportion of CD4. The proportion of CD4. Our findings show that IL-9 produced by helper T cells induces IL-8 expression, suggesting that IL-9 plays an important role in the development of endometriosis by stimulating IL-8 expression.

Topics: Adult; Endometriosis; Female; Humans; Interleukin-8; Interleukin-9; T-Lymphocytes, Helper-Inducer

Endometriosis is an inflammatory disease affecting reproductive-aged women. Immunologic disturbance, as well as inflammation, have crucial roles in the pathogenesis of endometriosis. In this study, we evaluated the effects of resveratrol treatment on expression of monocyte chemotactic protein-1 (MCP-1), interleukin-6 (IL-6), IL-8, and regulated upon activation, normal T cell expressed and secreted (RANTES) in endometrial stromal cells from patients with endometriosis compared with non-endometriotic controls. Thirteen eutopic (EuESCs) and nine ectopic (EESCs) endometrial stromal cells from endometriotic patients as well as eleven endometrial stromal cells from non-endometriotic controls (CESCs) were treated with resveratrol (100 μmol/L) or ethanol, and gene and/or protein expression of MCP-1, IL-6, IL-8 and RANTES was examined at 6, 24 and 48 hours following treatment in the cells from all origins. Resveratrol treatment significantly reduced gene and protein expression of MCP-1, IL-6, and IL-8 in EuESCs and EESCs compared with CESCs (P < .05-.001, P < .05-.001 and P < .05-<.01, respectively), and this reduction was more noticeable in EESCs than EuESCs (P < .05-<.001). Besides, resveratrol treatment significantly reduced RANTES protein expression in EESCs in all time intervals (P < .05). Resveratrol treatment significantly reduced the expression of MCP-1, IL-6, IL-8 and RANTES in EESCs.

Topics: Adult; Chemokine CCL2; Chemokine CCL5; Endometriosis; Female; Gene Expression Regulation; Humans; Interleukin-6; Interleukin-8; Middle Aged; Resveratrol; Stromal Cells; Young Adult

Endometriosis affects about 10-15% women for reproductive age, but it is not currently curable and the underlying etiology for this disease is still not clear. In the present study, functions and mechanisms of miR-182 and RELA in endometriosis were investigated. BAY 11-7082 was used to block NF-κB pathway. qRT-PCR, ELISA and western blot assays were employed to evaluate the expressions of miR-182 and RELA, inflammatory factors and epithelial-mesenchymal transition (EMT)-related markers, and activation of NF-κB pathway. MTT, wound healing or Transwell assays were used to evaluate the cell proliferation, migration and invasion capacities. Bioinformatic and dual-luciferase reporter assays were carried out to analyze the interaction between miR-182 and RELA. MiR-182 expression was decreased, while RELA was increased as developed from normal to eutopic and ectopic status, which was accompanied by upregulated inflammatory factors and EMT-related proteins. RELA was directly targeted by miR-182 in human endometrial stromal cells. Overexpression of RELA increased inflammation-associated and EMT-related markers expression, while miR-182 upregulation decreased the expression of these genes in a dose-dependent manner, which finally attenuated the proliferation, migration and invasion capacities of endometrial stromal cells through deactivation of NF-κB signaling pathway. Moreover, co-overexpression of RELA reversed the above effects induced by miR-182. In a word, miR-182 directly targeted RELA and inhibited proliferation, migration, invasion, EMT and inflammation of endometrial stromal cells through deactivation of NF-κB signaling pathway in endometriosis. These results provide new insights into the interaction between miR-182 and NF-κB pathway and their potential as therapeutic targets for treatment of endometriosis.

Topics: Cell Movement; Cell Proliferation; Computational Biology; Endometriosis; Endometrium; Epithelial Cells; Epithelial-Mesenchymal Transition; Female; Humans; Inflammation; Interleukin-6; Interleukin-8; MicroRNAs; Neoplasm Invasiveness; NF-kappa B p50 Subunit; Signal Transduction; Stromal Cells; Tetrazolium Salts; Thiazoles; Transcription Factor RelA; Transfection; Treatment Outcome; Wound Healing

Endometriosis is a common gynaecological disease of women in reproductive age, and is thought to arise from retrograde menstruation and implantation of endometrial tissue, mostly into the peritoneal cavity. The condition is characterized by a chronic, unresolved inflammatory process thereby contributing to pain as cardinal symptom in endometriosis. Elevated reactive oxygen species (ROS) and oxidative stress have been postulated as factors in endometriosis pathogenesis. We here set out for a systematic study to identify novel mechanisms and pathways relating to oxidative stress in ectopic peritoneal lesions. Using combined proteomic and transcriptomic approaches, we identified novel targets including upregulated pro-oxidative enzymes, such as amine oxidase 3/vascular adhesion protein 1 (AOC3/VAP1) as well as downregulated protective factors, in particular alkenal reductase PTGR1 and methionine sulfoxide reductase. Consistent with an altered ROS landscape, we observed hemoglobin / iron overload, ROS production and lipid peroxidation in ectopic lesions. ROS-derived 4-hydroxy-2-nonenal induced interleukin IL-8 release from monocytes. Notably, AOC3 inhibitors provoked analgesic effects in inflammatory pain models in vivo, suggesting potential translational applicability.

Topics: Aldehydes; Allyl Compounds; Amine Oxidase (Copper-Containing); Analgesics; Animals; Biomarkers; Cell Adhesion Molecules; Disease Models, Animal; Endometriosis; Female; Gene Expression Profiling; Heme; Humans; Inflammation Mediators; Interleukin-8; Iron; Lipid Peroxidation; Metabolic Networks and Pathways; Mice; Mice, Inbred BALB C; Myeloid Cells; Oxidative Stress; Peritoneal Diseases; Phagocytosis; Sulfonamides

What is the association between endometrioma-affected ovaries, their follicular fluid inflammatory microenvironment, and ovary-specific oocyte and embryo yield and quality?. Exposure-matched prospective cohort study conducted at a university-affiliated infertility clinic. Thirty-four women presenting for oocyte retrieval were enrolled between 2012 and 2013: women with unilateral endometrioma and no other observed peritoneal or deep lesions (n = 10) and women with no signs or symptoms of endometriosis (n = 24). Follicular fluid was aspirated at the time of oocyte retrieval. Samples from each ovary were analysed using a 27-plex immunoassay panel. The associations were evaluated by ovary-specific endometrioma exposure status (affected, unaffected, unexposed) with cytokine levels, oocyte yield and embryo quality.. Levels of interleukin (IL)-8 and monocyte chemoattractant protein-1 were higher in fluid obtained from endometrioma-affected ovaries compared with the unexposed ovaries from women without endometriosis, with intermediate levels observed in the contralateral unaffected ovaries. More modest differences were observed for IL-1β and IL-6. The affected ovaries of women with endometriosis yielded fewer oocytes (mean ± SD = 4.6 ± 2.3) compared with both the unaffected (6.0 ± 3.8) and unexposed (7.9 ± 5.6) ovaries. After adjusting for potential confounders and variables generated in a cytokine principal components analysis, oocyte yield remained slightly lower for the endometrioma-affected ovaries compared with unexposed ovaries. No informative differences among ovary groups for embryo quality parameters were observed.. The results suggest that the inflammatory milieu of ovarian endometriosis is strongly localized and has a more modestly systemic effect. The effect of endometriomas on infertility, however, cannot be entirely explained by increased inflammation.

Topics: Chemokine CCL2; Endometriosis; Female; Fertilization in Vitro; Follicular Fluid; Humans; Inflammation; Interleukin-8; Oocyte Retrieval; Oocytes; Ovarian Diseases; Ovary

Studies have shown a relationship between endometriosis and ovarian cancer. Our aims were to evaluate and compare the dosages of cytokines IL-2, IL-5, IL-6, IL-8, IL-10, and TNF-α in serum, intracystic fluid, and peritoneal fluid of patients with ovarian endometrioma, malignant and benign ovarian neoplasms, and non-neoplastic ovarian tumors; to verify if there is a correlation between the values of these cytokines between ovarian endometrioma and ovarian malignancy; and to determine the best cut-off point for serum cytokines that can be used to differentiate patients with ovarian malignancy and endometrioma.. The concentrations of cytokines were quantified by enzyme-linked immunosorbent assay (ELISA), analyzed by Kruskal-Wallis test with the Dunn post-test. Receiver operating feature (ROC) curve was used to obtain the area under the curve (AUC) and to determine the best cut-off values that could be used in the diagnosis of ovarian malignancy. Correlations of cytokine concentrations were performed by the Spearman test.. IL-6, IL-8, and IL-10 concentrations were higher in patients with malignant neoplasia. When evaluating the area under the curve (AUC) of serum cytokine levels comparing patients with malignant neoplasia and endometriomas, there was statistical significance for IL-6, IL-8, and IL-10.. Our results showed utility in serum concentrations of IL-6, IL-10, and IL-8 as parameters that differentiate endometriomas from ovarian malignancies.

Topics: Adolescent; Adult; Aged; Biomarkers, Tumor; Child; Diagnosis, Differential; Endometriosis; Female; Humans; Interleukin-10; Interleukin-6; Interleukin-8; Middle Aged; Neoplasms; Ovarian Neoplasms; ROC Curve; Young Adult

Concentrations of eight different cytokines and the level of expression of CD86 and CD163 macrophages were studied in peritoneal fluid in women with endometriosis. It was found that the concentration of both inflammatory (IL-6, IL-8, TNF-α) and anti-inflammatory cytokines (IL-4) as well as the level of macrophage expression of the proinflammatory marker CD86 and anti-inflammatory marker CD163 increased in women with mild external genital endometriosis (1-2 stage), and did not differ from the control group in women with severe endometriosis (3-4 stage). The content of IL-2, IL-10, CM-CSF and IFN-γ in the peritoneal fluid of women with endometriosis did not differ significantly from the control group. The results of the study indicate that the development of external genital endometriosis may be based on insufficient both inflammatory and anti-inflammatory activity of macrophages in the peritoneal fluid.. Issledovana kontsentratsiia vos'mi razlichnykh tsitokinov i uroven' ékspressii makrofagami CD86 i CD163 v peritoneal'noĭ zhidkosti u zhenshchin s éndometriozom. Ustanovleno, chto soderzhanie kak provospalitel'nykh (IL-6, IL-8, TNF-α), tak i protivovospalitel'nogo tsitokina (IL-4), a takzhe uroven' ékspressii makrofagami provospalitel'nogo markera CD86 i protivovospalitel'nogo markera CD163 v peritoneal'noĭ zhidkosti povyshaetsia u zhenshchin s legkimi formami naruzhnogo genital'nogo éndometrioza (1-2 st.); u zhenshchin s tiazhelymi formami éndometrioza (3-4 st.) issleduemye markery ne otlichaiutsia ot kontrol'noĭ gruppy. Soderzhanie IL-2, IL-10, CM-CSF i IFN-γ v peritoneal'noĭ zhidkosti u zhenshchin s éndometriozom ne imelo znachimykh razlichiĭ s kontrol'noĭ gruppoĭ. Rezul'taty issledovaniia svidetel'stvuiut o tom, chto v osnove razvitiia naruzhnogo genital'nogo éndometrioza mozhet lezhat' nedostatochnaia kak vospalitel'naia, tak i protivovospalitel'naia aktivnost' makrofagov v peritoneal'noĭ zhidkosti.

Topics: Ascitic Fluid; Cytokines; Endometriosis; Female; Genitalia; Humans; Interleukin-4; Interleukin-6; Interleukin-8; Macrophages; Tumor Necrosis Factor-alpha

Local pro-inflammatory environment and enhanced cell survival contribute to the endometriosis development. A serine/threonine kinase p38 mitogen-activated protein kinase (MAPK) mediates intracellular signaling of cytokine production, cell proliferation, and apoptosis in different cell types. The current study compares p38 MAPK activity in normal endometrium and endometriosis, and assesses role(s) of p38 MAPK on cytokine production and cell survival in endometriosis.. Immunohistochemical levels of total and phosphorylated (active) p38 MAPK as well as its correlation with interleukin 8 (IL-8) expression, and cell proliferation and apoptosis were compared in normal human endometrium and endometriosis. The action of p38 MAPK on pro-inflammatory cytokine-induced IL-8 and monocyte chemotactic protein (MCP)-1 expression in endometriotic cells were assessed by enzyme-linked immunosorbent assay. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide cell survival, 5-bromo-2'-deoxyuridine incorporation, and Terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick end labeling assays were used to determine the function of p38 MAPK in cultured human endometriotic stromal cell proliferation and apoptosis.. p38 MAPK activity was significantly higher in both eutopic and ectopic endometria compared to normal endometria during late proliferative and early secretory phases ( P < .05). Increased p38 MAPK activity in endometriotic cells correlated with IL-8 expression (Pearson correlation coefficient r = 0.83, P < .01), but not with apoptosis in vivo. The pro-inflammatory cytokines IL-1β and tumor necrosis factor (TNF)-α induced activation of p38 MAPK. Inhibition of p38 MAPK activity blocked IL-1β and TNF-α-induced IL-8 and MCP-1 secretion in cultured endometriotic stromal cells ( P < .05), but did not impact on endometriotic cell survival.. These results suggest that rather than modulating cell survival, increased p38 MAPK activity in endometriotic cells contributes to the pathogenesis of endometriosis by promoting the local inflammatory milieu.

Topics: Adult; Apoptosis; Cell Survival; Chemokine CCL2; Endometriosis; Endometrium; Female; Humans; Inflammation; Interleukin-1beta; Interleukin-8; p38 Mitogen-Activated Protein Kinases; Phosphorylation; Young Adult

CD74 is the primary receptor for macrophage migration inhibitory factor (MIF). Although expression of MIF has been described in endometriotic lesions, the cellular localization and function of the MIF receptor, CD74, are poorly understood. To further explore the role of CD74 in the pathophysiology of endometriosis, we utilized specimens from women with diagnostically confirmed endometriosis, women with no signs or symptoms of endometriosis (controls), and 8 baboons with experimentally induced endometriosis. Compared to eutopic endometrium from women with endometriosis, CD74 transcript expression was significantly increased in endometriotic lesion tissue. Similarly, cellular expression of CD74 was significantly greater in ectopic lesion tissue compared to paired eutopic endometrium, which both expressed greater CD74 expression compared to eutopic endometrium from control patients. Localization of CD74 was predominant to epithelial cells of ectopic and matched eutopic endometrium and was not influenced by the stage of the menstrual cycle. Eutopic endometrium from control patients did not express detectable levels of CD74 protein by immunohistochemistry. This pattern of expression and CD74 protein localization could be recapitulated in endometriotic lesion tissue from baboons with experimentally induced disease. Transfection of the endometriotic epithelial cell lines, 12Z with CD74 short hairpin RNA (shRNA), resulted in a significant decrease in CD74 protein expression, which was associated with a significant reduction in cellular proliferation as well as the expression of the prosurvival cytokine interleukin 8. Together, these data support the hypothesis that CD74 is elevated in endometriotic lesion tissue and may contribute to the pathogenesis of endometriosis by promoting cell survival.

Topics: Adult; Animals; Antigens, Differentiation, B-Lymphocyte; Cell Survival; Endometriosis; Endometrium; Epithelial Cells; Female; Histocompatibility Antigens Class II; Humans; Interleukin-8; Middle Aged; Papio anubis; Receptors, Immunologic; Species Specificity; Young Adult

Endometriosis is a chronic gynecological disorder defined as the presence of endometrial tissue within extra-uterine sites. The primary symptoms are infertility and chronic pain. The inflammatory environment and aberrant immune responses in women with endometriosis may be directly associated with the initiation and progression of endometriotic lesions. In the present study, the secretion of inflammatory cytokines was evaluated in cultures of primary endometrial cells (ECs) isolated from the endometrium of women with and without endometriosis. The presence of endometriotic cells leads to alterations in the secretory profile of healthy ECs. The expression of the inflammatory cytokines interleukin (IL)‑6 and IL‑8 was significantly increased in endometriotic and co‑cultured cells compared with healthy ECs. IL‑6 expression was strongly correlated with IL‑8 expression in endometriotic cells. IL‑1β expression was increased on day 10 of co‑culture to 48.30 pg/ml and may be associated with the long‑term co‑culture, rather than IL‑6 and IL‑8 expression. IL‑6 expression was strongly correlated with cell number, whereas IL‑8 expression was moderately correlated with cell number. Additionally, it was observed that co‑cultured cells exhibited a different population of cells, with expression of the mesenchymal stem cell marker cell surface glycoprotein MUC18, indicating a putative role of endometrial mesenchymal stem cells in the secretion of cytokines and disease development. These results indicate a predominant role of primary endometriotic cells in the secretion of cytokines, which contributes to the disrupted peritoneal and endometrial environment observed in the women with endometriosis.

Topics: Adolescent; Adult; CD146 Antigen; Coculture Techniques; Endometriosis; Female; Gene Expression Profiling; Gene Expression Regulation; Humans; Inflammation; Interleukin-6; Interleukin-8; Middle Aged

Diseases are complex systems that can be studied through the integration of data derived from different disciplines to obtain a global and reliable picture of the biological phenomenon under investigation. Based on the recent observations that the metabolomics profiling of follicular fluids reflects the ovarian microenvironment of women and that endometriosis represents an example of complex diseases, clearly diagnosed by laparoscopy, we thought that the follicular fluids of endometriosis patients can represent a study model to evaluate the possibility of integrating data obtained by different approaches. Hence, the aim of this work was to analyze and integrate different clinical chemistry parameters with specific reference to the metabolic profile, inflammatory state and cell damage by a

Topics: Adult; Chemokine CCL11; Chemokine CXCL10; Endometriosis; Female; Follicular Fluid; Humans; Insulin; Interleukin-8; Isoenzymes; L-Lactate Dehydrogenase; Lactic Acid; Metabolomics; Phospholipids; Vascular Endothelial Growth Factor A

The study aimed to explore the type 1 and type 2 cytokines expression in the endometrium from women affected by endometriosis compared to controls. The expression of TSG-6, a multifunctional protein involved in several inflammatory disease, was also evaluated. Study Design SETTING: Experimental clinical study.. 10 patients affected by endometriosis and 11 controls.. Patients underwent to an ultrasound transvaginal examination and a diagnostic hysteroscopy in order to exclude any uterine abnormality. All patients underwent endometrial biopsy using a Novak's curette.. The endometrial expression of type 1 (IL- 1 β TNF-α, IL-8) and type 2 (IL-10) cytokines, and of TSG-6 was evaluated by immunohistochemistry and by real time PCR. The expression of TSG-6 was confirmed by western blot.. Results of PCR analysis and of immunohistochemistry revealed an increased expression of IL-1β, TNF-α, IL-8 and of TSG-6 in the endometrium of endometriosic patients. IL-10 expression did not show any difference.. An increased expression of pro-inflammatory type 1 cytokines was demonstrated in the endometrium from endometriosic patients, suggesting an endometrial environment harmful for implantation due to the prevalence of Th1 related immunity. An increased expression of TSG-6 was also demonstrated for the first time. Our findings concur to better define the inflammatory imbalance and the abnormal endometrial receptivity, reported in literature, of the eutopic endometrium of women affected by endometriosis.

Topics: Adult; Blotting, Western; Case-Control Studies; Cell Adhesion Molecules; Endometriosis; Endometrium; Female; Gene Expression Regulation; Humans; Infertility, Female; Inflammation Mediators; Interleukin-10; Interleukin-1alpha; Interleukin-1beta; Interleukin-8; Real-Time Polymerase Chain Reaction; RNA, Messenger; Tumor Necrosis Factor-alpha; Ultrasonography; Young Adult

Endometriosis is a debilitating condition that is categorized by the abnormal growth of endometrial tissue outside the uterus. Although the pathogenesis of this disease remains unknown, it is well established that endometriosis patients exhibit immune dysfunction. Interleukin (IL)-33 is a danger signal that is a critical regulator of chronic inflammation. Although plasma and peritoneal fluid levels of IL-33 have been associated with deep infiltrating endometriosis, its contribution to the disease pathophysiology is unknown. We investigated the role of IL-33 in the pathology of endometriosis using patient samples, cell lines and a syngeneic mouse model. We found that endometriotic lesions produce significantly higher levels of IL-33 compared to the endometrium of healthy, fertile controls. In vitro stimulation of endometrial epithelial, endothelial and endometriotic epithelial cells with IL-33 led to the production of pro-inflammatory and angiogenic cytokines. In a syngeneic mouse model of endometriosis, IL-33 injections caused systemic inflammation, which manifested as an increase in plasma pro-inflammatory cytokines compared to control mice. Furthermore, endometriotic lesions from IL-33 treated mice were highly vascularized and exhibited increased proliferation. Collectively, we provide convincing evidence that IL-33 perpetuates inflammation, angiogenesis and lesion proliferation, which are critical events in the lesion survival and progression of endometriosis.

Topics: Animals; Ascitic Fluid; Cell Line; Cell Line, Tumor; Cytokines; Disease Models, Animal; Endometriosis; Endometrium; Female; Human Umbilical Vein Endothelial Cells; Humans; Inflammation; Interleukin-33; Interleukin-8; Mice; Mice, Inbred C57BL; Neovascularization, Pathologic

This study analyzed whether trefoil factor 3 (TFF3) is locally elevated and correlated with common biomarkers and inflammatory processes in endometriosis. Peritoneal fluid (PF) was obtained from 50 women and serum from 124 women with or without endometriosis. Experimental endometriosis was induced in female C57BL/6 mice by syngeneic transplantation of uterine tissue to the abdominal wall. Levels of TFF3 in PF of women with endometriosis were significantly increased ( P < .05) and correlated with local levels of known biomarkers for endometriosis: cancer antigen (CA) 125, CA-19-9, interleukin 8, monocyte chemotactic protein 1, and matrix metalloproteinase 7. Serum levels of TFF3 in women were significantly influenced by the menstrual cycle but were independent from disease state. In mice, local TFF3 levels were significantly elevated in early endometriosis (up to 4 weeks after transplantation, P < .001) and corresponded to increases in spleen weight as marker for systemic inflammation. This study provides the first evidence that TFF3 is locally elevated in the peritoneal cavity in endometriosis and might play a role in disease pathogenesis and its associated inflammatory processes. Furthermore, the results show that TFF3 is regulated through the menstrual cycle. With respect to animal models, syngeneic mouse model does reflect local TFF3 upregulation in the peritoneal cavity affected by endometriosis.

Topics: Adult; Animals; Ascitic Fluid; Biomarkers; Chemokine CCL2; Endometriosis; Female; Humans; Interleukin-8; Matrix Metalloproteinase 7; Menstrual Cycle; Mice; Peritoneal Cavity; Trefoil Factor-3

The objective was to evaluate the effect of lignocaine on cytokine expression and secretion in vitro in peritoneal fluid macrophages and endometriotic stromal cells.. Experimental in vitro study on human cells.. Peritoneal fluid (n = 10) and samples from endometriotic cysts (n = 7) were collected from 13 women (women with endometriosis n = 8, and healthy controls n = 5) during surgery for clinical reasons.. Macrophages from the peritoneal fluid and cells from the inside of the endometriotic cysts capsules were isolated and cultivated for 24 to 48 hours in medium with and without the supplement of lignocaine 0.1 or 1.0 mg/mL. Relative gene expression of monocyte chemotactic protein 1 (MCP-1), interleukin 6 (IL-6), and IL-8 was evaluated with quantitative polymerase chain reaction and compared between treated and untreated cells with Wilcoxon matched pairs. The concentrations of MCP-1, IL-6, and IL-8 were measured using enzyme-linked immunosorbent assay and were compared between treated and untreated cells with Wilcoxon matched pairs.. The gene expression and protein secretion of IL-8 in endometriotic stromal cells after incubation with lignocaine 0.1 mg/mL were significantly decreased after 24 hours compared to the controls ( P = .028 and P = .018). Macrophages from healthy controls had a significant lower gene expression of all tested cytokines ( P = .043) after treatment with lignocaine, but there were no significant differences in protein level. Macrophages from women with endometriosis showed diverging results since 3 of 5 samples showed increased gene expression of 1 (n = 2) or 2 cytokines (n = 1) after lignocaine treatment.. Lignocaine can affect the gene expression and secretion of some proinflammatory cytokines in vitro.

Topics: Adult; Anesthetics, Local; Ascitic Fluid; Cells, Cultured; Chemokine CCL2; Endometriosis; Endometrium; Female; Humans; Interleukin-6; Interleukin-8; Lidocaine; Macrophages, Peritoneal; Stromal Cells

Endometriosis (EM) is a hormone-dependent chronic inflammatory disease, usually accompanied by a high level of localized estrogen and abnormal levels of cytokines, which are regulated by GATA-3 in lymphocytes. This study aimed to investigate the role of estrogen on GATA-3 expression and the relationship between GATA-3 and cytokine response.. Endometrial tissues collected from 20 patients who underwent laparoscopic or open surgery were used. Immunohistochemistry, quantitative polymerase chain reaction, Western blot analysis, cell transfection, estrogen treatments and enzyme-linked immunosorbent assays were performed to evaluate the effects of estrogen on GATA-3 expression and the relationship between estrogen-induced GATA-3 and the Th2 immune status of EM.. Estrogen regulated the expression of GATA-3 in a dose and time-dependent manner. GATA-3 was relocated from the cytoplasm to the nucleus. Estrogen and GATA-3 regulated Th2 cytokine expression in eutopic endometrial cells, including interleukin (IL)-6, IL-8 and IL-10. Moreover, interferon-γ and IL-2 were highly expressed in the GATA-3 knockdown groups.. In summary, GATA-3 was induced by estrogen and may promote the occurrence and development of EM by regulating the secretion of cytokines in the eutopic endometrial cells of EM patients.

Topics: Adult; Cell Nucleus; Cells, Cultured; Cytokines; Cytoplasm; Endometriosis; Estrogens; Female; GATA3 Transcription Factor; Humans; Interleukin-10; Interleukin-6; Interleukin-8; Middle Aged; RNA, Messenger; Th2 Cells

Endometriosis is an estrogen-dependent, chronic inflammatory disease. Recent studies have shown that vitamin D (VD) is an effective modulator of the immune system and plays an important role in controlling many inflammatory diseases.. The objective of the study was to clarify the in vitro effects of 1,25-dihydroxy vitamin D3 (1,25[OH]2D3) on human endometriotic stromal cells (ESCs) and to determine the serum levels of VD in endometriosis patients.. ESCs were isolated from ovarian endometrioma and cultured with 1,25(OH)2D3. Gene expression of IL-8, cyclooxygenase-2, microsomal prostaglandin E synthase-1, microsomal prostaglandin E synthase-2, cytosolic prostaglandin E synthase, 15-hydroxyprostaglandin dehydrogenase, matrix metalloproteinase (MMP)-2, and MMP-9 was examined using quantitative RT-PCR. The production of IL-8 and prostaglandin E2 was measured using an ELISA and an enzyme immunoassay. Viable cell number was assessed using a cell-counting assay, and DNA synthesis was assessed using the bromodeoxyuridine incorporation assay. Apoptosis was assessed using flow cytometry. The expression of inhibitory-κBα protein was detected using Western blotting. The serum levels of 25-hydroxyvitamin D3 and 1,25(OH)2D3 were measured by a RIA.. In vitro studies showed that 1,25(OH)2D3 significantly reduced IL-1β- or TNF-α-induced inflammatory responses, such as IL-8 expression and prostaglandin activity. 1,25(OH)2D3 also reduced viable ESC numbers and DNA synthesis but did not affect apoptosis. MMP-2 and MMP-9 expressions were reduced by 1,25(OH)2D3. 1,25(OH)2D3 inhibited nuclear factor-κB activation. The serum 25-hydroxyvitamin D3 levels were significantly lower in women with severe endometriosis than in the controls and women with mild endometriosis. Serum 1,25(OH)2D3 levels were not different between groups.. VD modulates inflammation and proliferation in endometriotic cells, and a lower VD status is associated with endometriosis. Taken together, VD supplementation could be a novel therapeutic strategy for managing endometriosis.

Topics: Adult; Apoptosis; Calcitriol; Cell Proliferation; Cells, Cultured; Cyclooxygenase 2; Endometriosis; Endometrium; Female; Gene Expression; Humans; Interleukin-8; Matrix Metalloproteinase 2; Matrix Metalloproteinase 9; Ovarian Diseases; Prostaglandin-E Synthases; Stromal Cells; Vitamin D

The study aimed to investigate key intrafollicular prognostic factors among various cytokines and angiogenic molecules for prediction of mature oocytes and good-quality embryos in women with endometriosis undergoing in vitro fertilization (IVF).. Paired follicular fluid and serum samples were collected from 200 women with advanced stage endometriosis and 140 normal ovulating women during oocyte retrieval. The concentrations of cytokines (pro-inflammatory: IL-1β, TNF-α, IL-2, IL-8, IL-12, IFN-γ; anti-inflammatory: IL-4, IL-6, IL-10) and angiogenic molecules (vascular endothelial growth factor (VEGF), adrenomedullin, angiogenin) were determined in follicular fluid and serum using ELISA. Expression of these molecules was subjected to multivariate analysis for the identification of major predictive markers of oocyte and embryo quality. Receiver operating characteristic (ROC) curve was applied to determine the best cutoff point for the discrimination between mature and immature oocytes in these women.. Significant increases in levels of cytokines and angiogenic molecules were observed in women with endometriosis compared to controls (P < 0.001). From the validated partial least squares-discriminant analysis (PLS-DA) model, IL-8, IL-12, and adrenomedullin were identified as the most important factors contributing to endometriosis and were negatively associated with oocyte maturity and embryo quality.. The levels of IL-8, IL-12, and adrenomedullin may be good indicators of embryo and oocyte quality in endometriosis patients undergoing IVF. Further studies are necessary to ascertain the potential of these markers for oocyte and embryo developmental competence which may help improve the chances of a successful IVF in endometriosis patients.

Topics: Adrenomedullin; Adult; Embryo Transfer; Endometriosis; Female; Fertilization in Vitro; Follicular Fluid; Humans; Infertility, Female; Interleukin-12; Interleukin-8; Oocyte Retrieval; Oocytes

Previous study indicated that bleeding into the peritoneum may accelerate inflammatory response in endometriosis-like grafts in mice. To identify changes in protein levels in the grafts from mice that underwent unilateral ovariectomy (uOVX), which causes bleeding from ovarian arteries and vein, the grafts were generated by injecting a suspension of human endometrial cells in BALB/c nude female mice, and protein profile changes were compared with non-uOVX control mice. The level of α1-antitrypsin (α1-AT) decreased in grafts from nude mice that underwent uOVX. The levels of phosphorylated Akt, mammalian target of rapamycin, S6K, regulatory factors for cell survival, and of phosphorylated nuclear factor κB, an inflammatory mediator, were higher in endometriosis-like grafts from the uOVX group than from the control. The grafts were mostly comprised of stromal cells. The bioactivity of α1-AT was assessed by investigating cytokine expression in protease-activated receptor (PAR) 1/2 agonists-stimulated stromal cells. The PARs promoted the expression of interleukin 8 (IL-8), but treatment with α1-AT blocked IL-8 expression dose dependently. Knocking down α1-AT expression increased the constitutive IL-6, IL-8, and cyclooxygenase 2 expression as well as PAR1 agonist-stimulated IL-6 expression. These findings support the notion that decreased α1-AT protein in the grafts constituted with human endometrial cells in mice may have exacerbated inflammation in endometriosis-like grafts, suggesting the possible involvement of α1-AT in the pathophysiology of endometriosis.

Topics: alpha 1-Antitrypsin; Animals; Cells, Cultured; Cyclooxygenase 2; Disease Models, Animal; Endometriosis; Endometrium; Female; Heterografts; Humans; Interleukin-6; Interleukin-8; Mice, Inbred BALB C; Mice, Nude; NF-kappa B; Ovariectomy; Phosphorylation; Proto-Oncogene Proteins c-akt; Receptor, PAR-1; Receptor, PAR-2; Ribosomal Protein S6 Kinases, 70-kDa; RNA Interference; Signal Transduction; TOR Serine-Threonine Kinases; Transfection

Endometriosis is a chronic, inflammatory disease characterized by the growth of endometrial tissue in aberrant locations outside the uterus. Neoangiogenesis or establishment of new blood supply is one of the fundamental requirements of endometriotic lesion survival in the peritoneal cavity. IL-17A is emerging as a potent angiogenic and proinflammatory cytokine involved in the pathophysiology of several chronic inflammatory diseases such as rheumatoid arthritis and psoriasis. However, sparse information is available in the context of endometriosis. In this study, we demonstrate the potential importance of IL-17A in the pathogenesis and pathophysiology of endometriosis. The data show a differential expression of IL-17A in human ectopic endometriotic lesions and matched eutopic endometrium from women with endometriosis. Importantly, surgical removal of lesions resulted in significantly reduced plasma IL-17A concentrations. Immunohistochemistry revealed localization of IL-17A primarily in the stroma of matched ectopic and eutopic tissue samples. In vitro stimulation of endometrial epithelial carcinoma cells, Ishikawa cells, and HUVECs with IL-17A revealed significant increase in angiogenic (vascular endothelial growth factor and IL-8), proinflammatory (IL-6 and IL-1β), and chemotactic cytokines (G-CSF, CXCL12, CXCL1, and CX3CL1). Furthermore, IL-17A promoted tubulogenesis of HUVECs plated on Matrigel in a dose-dependent manner. Thus, we provide the first evidence, to our knowledge, that endometriotic lesions produce IL-17A and that the removal of the lesion via laparoscopic surgery leads to the significant reduction in the systemic levels of IL-17A. Taken together, our data show a likely important role of IL-17A in promoting angiogenesis and proinflammatory environment in the peritoneal cavity for the establishment and maintenance of endometriosis lesions.

Topics: Adult; Cell Line; Chemokine CX3CL1; Chemokine CXCL1; Chemokine CXCL12; Endometrial Neoplasms; Endometriosis; Endometrium; Female; Granulocyte Colony-Stimulating Factor; Human Umbilical Vein Endothelial Cells; Humans; Inflammation; Interleukin-17; Interleukin-1beta; Interleukin-6; Interleukin-8; Laparoscopy; Neovascularization, Pathologic; Vascular Endothelial Growth Factor A; Young Adult

Chronic inflammation is important for the occurrence of endometriosis, but the molecular mechanisms are still poorly understood. TLR4 is not only expressed on immune cells but is also present in the human endometrium, and its regulation might be crucial for the pathogenesis of endometriosis.. In this study, the expression of TLR4 in normal, eutopic endometrium, and ectopic tissues was analyzed by immunohistochemistry. The expression of the key molecules in endometrial stromal cells (ESCs) was assessed by in-cell Western assays. The invasion of eutopic ESCs from patients with endometriosis was evaluated by Matrigel invasion assay. The effects of CXCL8 on the proliferation of ESCs in vitro were assessed using BrdU assays.. We found that the expression of TLR4 is higher in the eutopic endometrium than the normal endometrium and that ectopic tissue had the highest level of expression. TLR4 activation stimulated IL-8 secretion and the expression of its receptor CXCR1 in ESCs by activating p38/ERK, but not JNK and NK-κB signal pathways. IL-8 could enhance the invasion and proliferation of ESCs through the FAK signal pathway, and these effects could be abolished by an anti-CXCL8 neutralizing antibody or by a FAK inhibitor.

Topics: Adult; Autocrine Communication; Cell Movement; Cell Proliferation; Cells, Cultured; Choristoma; Endometriosis; Endometrium; Female; Focal Adhesion Kinase 1; Humans; Interleukin-8; Stromal Cells; Toll-Like Receptor 4; Young Adult

Angiogenesis and inflammation are pivotal processes in developing endometriosis in the peritoneal cavity. The aim of the present study was to evaluate these two processes in women with endometriosis who had been treated with danazol to determine the sensitivity of a non-invasive test in diagnosing endometriosis. The clinical follow-up study was conducted in a group of 103 women diagnosed laparoscopically with endometriosis. Thirty-five patients qualified for danazol treatment. Pain was assessed using a visual analogue scale, whereas endometriosis was assessed using the revised American Society of Reproductive Medicine (rASRM) scale. Cancer antigen (CA)-125 and C-reactive protein (CRP) concentrations in plasma and peritoneal fluid were determined by immunoenzymatic methods, whereas vascular endothelial growth factor (VEGF) and interleukin (IL)-1β concentrations in plasma and peritoneal fluid were determined by ELISA. Endometrial expression of IL-8 and platelet-derived growth factor alpha polypeptide (PDGF-A) was determined using real-time polymerase chain reaction (PCR). Women with endometriosis (68.9% of patients) had higher plasma concentrations of CA-125, as well as higher concentrations of both CA-125 and VEGF in the peritoneal fluid. Endometrial expression of IL-8 mRNA was significantly higher, whereas that of PDGF-A was significantly lower, in contrast. After danazol treatment the patients reported lower pain scores; in addition, CA-125 concentrations in the plasma were decreased (P<0.001), whereas VEGF concentration in the plasma increased (P=0.009). For the diagnosis of endometriosis, none of the combinations of given markers had a sensitivity >60%. Danazol treatment is highly effective in relieving pain and decreasing CA-125 concentrations in the plasma. Higher plasma concentrations of VEGF after treatment could imply stimulation of angiogenesis.

Topics: Ascitic Fluid; Biomarkers; C-Reactive Protein; CA-125 Antigen; Danazol; Endometriosis; Enzyme-Linked Immunosorbent Assay; Female; Follow-Up Studies; Humans; Immunoenzyme Techniques; Inflammation; Interleukin-1beta; Interleukin-8; Neovascularization, Pathologic; Pain Measurement; Platelet-Derived Growth Factor; Real-Time Polymerase Chain Reaction; Sensitivity and Specificity; Vascular Endothelial Growth Factor A

Endometriosis is a chronic inflammatory disease. Sirtuin 1 (SIRT1) plays a role in regulation of inflammation. The role of SIRT1 in endometriosis remains unknown. We here addressed the anti-inflammatory effects of SIRT1 on endometriosis.. The expression of SIRT1 in human ovarian endometriomas and eutopic endometria were examined using immunohistochemistry and reverse transcription polymerase chain reaction (RT-PCR). Endometriotic stromal cells (ESC) obtained from endometriomas were exposed to either resveratrol or sirtinol, an activator or inhibitor of sirtuins, respectively, and tumor necrosis factor (TNF)-α-induced interleukin (IL)-8 release from the ESC was assessed at mRNA and protein levels.. Both immunochemistry and RT-PCR demonstrated that SIRT1 was expressed in ESC and normal endometrial stromal cells. Resveratrol suppressed TNF-α-induced IL-8 release from the ESC in a dose-dependent manner while sirtinol increased IL-8 release.. These opposing effects of SIRT1-related agents suggest that IL-8 release from the ESC is modulated through the SIRT1 pathway. Resveratrol may have the potential to ameliorate local inflammation in endometriomas.

Topics: Adult; Anti-Inflammatory Agents, Non-Steroidal; Benzamides; Cells, Cultured; Endometriosis; Endometrium; Enzyme Activators; Enzyme Inhibitors; Female; Humans; Interleukin-8; Naphthols; Ovarian Diseases; Ovary; Resveratrol; Signal Transduction; Sirtuin 1; Stilbenes; Stromal Cells

How is the tumor necrosis factor (TNF) α-induced inhibitor of apoptosis (IAP) protein expression in endometriotic stromal cells (ESCs) involved in cell viability and signaling pathways?. Endometriotic stromal cells were isolated from ovarian chocolate cysts in 20 patients who underwent laparoscopic surgery. IAP protein expression and IκB phosphorylation were evaluated by Western blot analysis. Interleukin (IL)-8 protein expression and cell proliferation were assessed by ELISA.. Cellular IAP (cIAP)-2 protein expression in endometriotic tissue was higher than that of endometrium. TNFα markedly enhanced cIAP-2 protein expression in ESCs. Pretreatment with a nuclear factor (NF)-κB inhibitor attenuated TNFα-induced cIAP-2 expression. An antagonist of IAPs abrogated TNFα-induced cIAP-2 protein expression and showed a decrease in TNFα-induced IL-8 protein expression and BrdU incorporation in ESCs.. TNFα and its downstream NFκB pathway have proven to be critical regulators of highly expressed cIAP-2 in ESCs. cIAP-2 may be a novel therapeutic target for endometriosis.

Topics: Cell Proliferation; Cells, Cultured; Endometriosis; Endometrium; Female; Gene Expression Regulation; Humans; I-kappa B Proteins; Inhibitor of Apoptosis Proteins; Interleukin-8; Molecular Targeted Therapy; NF-kappa B; Oligopeptides; Phosphorylation; Signal Transduction; Stromal Cells; Tosylphenylalanyl Chloromethyl Ketone; Tumor Necrosis Factor-alpha

It has reported that human endometrial stromal cells (ESCs) express thymic stromal lymphopoietin (TSLP), and TSLP concentrations in the serum and peritoneal fluid were higher in women with endometriosis. Endometriosis is an estrogen-dependent disease. The present study aimed to elucidate whether and how estrogen regulates the growth of ESCs through TSLP. The ESCs behaviors in vitro were verified by SRB assay and Ki67 level detection, respectively. In addition, the effects of estrogen on TSLP and TSLP on the correspondent functional molecules were investigated by ELISA and flow cytometry. Here we found that estrogen stimulated the secretion of TSLP in a dosage-dependent manner. Recombinant human TSLP stimulates the secretion of MCP-1 and IL-8, and markedly promotes the viability and proliferation relative gene Ki-67 expression of ESCs. These effects could be abolished by the inhibitor for JNK or NF-κB signal, respectively. Moreover, not only anti-TSLP neutralizing antibody, but also blocking JNK or NF-κB signal by inhibitor abrogated the stimulatory role in the production of MCP-1 and IL-8, and the growth of ESCs induced by estrogen. Our current study has demonstrated that TSLP is involved in the regulation of estrogen on the secretion of MCP-1 and IL-8, and the growth of ESCs through JNK and NF-κB signal pathways, which suggests that the abnormal high expression of TSLP induced by estrogen may play an important role in ESCs growth and finally contribute to the origin and development of endometriosis.

Topics: Adult; Cell Proliferation; Cell Survival; Cells, Cultured; Chemokine CCL2; Cytokines; Dose-Response Relationship, Drug; Endometriosis; Endometrium; Estradiol; Female; Humans; Interleukin-8; JNK Mitogen-Activated Protein Kinases; Ki-67 Antigen; Middle Aged; NF-kappa B; Ovarian Diseases; Protein Kinase Inhibitors; Signal Transduction; Stromal Cells; Thymic Stromal Lymphopoietin; Up-Regulation; Young Adult

Immune tolerance to endometriotic cells is important to promote endometriosis. Tryptophan 2,3-dioxygenase (TDO) enhances immune tolerance by catabolizing tryptophan to kynurenine. We studied whether interleukin-1β (IL-1β), a typical endometriosis-associated cytokine, affects the expression of TDO and the catabolism of tryptophan in endometrioma stromal cells (ESCs). We also studied whether the expression of TDO is involved in IL-1β-induced secretion of IL-6 and IL-8 in ESCs.. Nineteen endometriotic patients of reproductive age with normal menstrual cycles were recruited. Primary cultures of ESCs were treated with IL-1β and TDO siRNA. TDO mRNA was measured using quantitative PCR. TDO protein was measured using Western blotting. Concentrations of kynurenine in condition media were measured using Ehrlich reagent. Concentrations of tryptophan in conditioned media were measured using tryptophan detection kit. Concentrations of IL-6 and IL-8 in conditioned media were measured using ELISA kits.. IL-1β (1 ng/mL) increased the expression of TDO mRNA and TDO protein in ESCs. IL-1β-treated ESCs increased the production of kynurenine and the effect was inhibited by TDO siRNA. Treatment with the siRNA also decreased IL-1β-induced secretion of IL-6 and IL-8 from ESCs.. IL-1β is suggested to stimulate tryptophan catabolism and production of IL-6 and IL-8 by increasing TDO expression in endometriosis.

Topics: Adult; Cells, Cultured; Endometriosis; Endometrium; Female; Gene Expression Regulation, Enzymologic; Humans; Interleukin-1beta; Interleukin-6; Interleukin-8; Stromal Cells; Tryptophan Oxygenase

How does hypoxia-mediated down-regulation of dual specificity phosphatase-2 (DUSP2) promote endometriotic lesion development?. Inhibition of DUSP2 by hypoxia enhances endometriotic lesion growth via promoting interleukin-8 (IL-8)-dependent angiogenesis.. Angiogenesis is a prerequisite for the development of endometriosis. DUSP2 is down-regulated in endometriotic stromal cells in a hypoxia inducible factor-1α-dependent manner. Down-regulation of DUSP2 contributes to the pathological process of endometriosis.. A laboratory study recruiting 20 patients of reproductive age with endometriosis and normal menstrual cycles, and an autoimplant-induced mouse model of endometriosis using 13 mice in a 28-day treatment.. IL-8 mRNA levels were assayed in endometrial stromal cells maintained in normoxic or hypoxic (1% O2) conditions, with or without DUSP2 knockdown. Promoter activity and chromatin immunoprecipitation (ChIP) assays were conducted to characterize the regulation of IL-8 by DUSP2. Conditioned media from cells maintained in normoxic or hypoxic conditions, and cells with/without DUSP2 knockdown were collected to investigate the angiogenic capacity using an in vitro tube formation assay. Reparixin, an IL-8 receptor blocker, was administered to investigate the role of IL-8 in hypoxia-mediated angiogenesis and the development of endometriotic-like lesions in an autotransplanted mouse model.. IL-8 mRNA was increased by both hypoxia and DUSP2 knockdown in endometrial stromal cells in an extracellular signal-regulated protein kinase-dependent manner (P < 0.05 versus control). Promoter activity and ChIP assays demonstrated that expression of IL-8 was regulated by CCAAT/enhancer binding protein α (P < 0.05 versus control). Furthermore, conditioned media collected from hypoxia-exposed or DUSP2 knockdown endometrial stromal cells promoted tube formation, which was abolished by co-treatment with reparixin (P < 0.05 versus control). Results from the autotransplanted mouse model demonstrated that number of blood vessels and size of endometriotic-like lesions were markedly reduced in recipient mice treated with reparixin (P < 0.05 versus control).. This study was conducted in primary human cell cultures and a mouse model, therefore may not fully reflect the situation in vivo.. This is the first study to highlight the potential application of an IL-8 receptor blocker as a therapeutic target to treat endometriosis. This study demonstrates IL-8 as a key angiogenic factor regulated by hypoxia/DUSP2, which suggests an alternative mechanism through which hypoxia may promote angiogenesis.. This study was funded by the National Science Council of Taiwan (NSC101-2314-B-006-043-MY2). The author declares that there is no conflict of interest.

Topics: Animals; Cell Hypoxia; Dual Specificity Phosphatase 2; Endometriosis; Female; Gene Knockdown Techniques; Humans; Interleukin-8; Mice; Neovascularization, Pathologic; Signal Transduction; Up-Regulation

Do endometriomas induce an inflammatory reaction with increased cytokine concentrations in nearby follicles and thereby affect follicular development during controlled ovarian stimulation for in vitro fertilization (IVF)?. With most endometriomas, there is no evidence of increased cytokine concentrations in the ipsilateral leading follicle. Infrequently, the concentration of inflammatory cytokines is increased in the follicular fluid (FF) and associated with diminished ovarian response.. The link between peritoneal endometriosis, inflammation and infertility is well established; however, the association between intraovarian inflammation and endometrioma is unknown.. This prospective cohort study included 117 infertile women undergoing IVF in a tertiary infertility clinic at Oslo University Hospital Rikshospitalet, Norway, during the period May 2009 to September 2011.. There were 47 patients with unilateral endometrioma and 17 patients with bilateral endometrioma, while the 53 control patients had unexplained or male factor infertility. Concentrations of IL-1β, IL-6, IL-8, IL-10, IL-12 and TNF-α were measured in serum and in the fluid of the largest pre-ovulatory follicles from each ovary of each participant.. Cytokine levels in the follicular fluid from the two ovaries in women with unilateral endometriomas were comparable, and were not significantly altered compared with that of control groups with male factor infertility, unexplained infertility or bilateral endometriomas. Compared with serum levels, the follicular fluid levels of IL-8 and IL-6 were higher, suggesting a local production or recruitment. The follicular fluid IL-8 level varied considerably and showed an inverse relationship with IL-12, IL-10 and TNF-∝, suggesting a complex interaction between various immune cells. A small group of patients (n = 3) had increased levels of all follicular fluid cytokines combined with moderately to slightly elevated serum levels and these patients had a significantly lower ovarian response.. For ethical reasons, the endometriomas were diagnosed indirectly by ultrasound rather than by histology.. This paper reveals that endometriomas seldom induce inflammation in nearby follicles during IVF; therefore, routine cystectomy prior to IVF may not be necessary. Cytokine levels in the follicular fluid, nonetheless, show distinctive patterns and increased levels may be linked to reduced ovarian response independent of the cause of infertility.

Topics: Cohort Studies; Endometriosis; Female; Fertilization in Vitro; Follicular Fluid; Humans; Interleukin-10; Interleukin-12; Interleukin-1beta; Interleukin-6; Interleukin-8; Interleukins; Ovarian Follicle; Tumor Necrosis Factor-alpha

To evaluate the effects of parthenolide on human endometriotic cells and murine endometriotic lesions.. Experimental study.. University hospital and laboratory of animal science.. Twenty women with ovarian endometrioma and 30 mice.. Ectopic endometrial tissue from the endometrioma was collected.. Human endometriotic stromal cells (ESCs) were pretreated with parthenolide and exposed to tumor necrosis factor (TNF)-α. Interleukin 8 (IL-8) and COX-2 gene expressions were evaluated by real-time reverse transcription-polymerase chain reaction. Interleukin-8 protein, prostaglandin E₂ (PGE₂) level, and intranuclear p65 protein concentration were determined by ELISA. Cell proliferation was assessed by 5-bromo-2'-deoxyuridine-ELISA. Phosphorylation of signaling pathways in ESCs was evaluated by Western blotting. Gene expression and proliferative activity in murine endometriosis-like lesions were assessed by real-time reverse transcription-polymerase chain reaction and Ki67 staining, respectively.. With parthenolide pretreatment, TNF-α-induced IL-8 gene and protein expression in ESCs were diminished. Tumor necrosis factor α-induced COX-2 expression and PGE2 synthesis were also inhibited. Adding parthenolide repressed TNF-α-induced 5-bromo-2'-deoxyuridine incorporation and IκB phosphorylation in ESCs. As in vivo experiments, administering parthenolide reduced the number, surface area, and weight, the level of Vegf, Il-6, Mcp-1, and Lif gene expression, and the percentage of Ki67-positive cells in murine endometriosis-like lesions.. Parthenolide repressed the development of endometriosis by suppressing the inflammatory peritoneal environment through the nuclear factor κB pathway.

Topics: Animals; Anti-Inflammatory Agents; Blotting, Western; Cell Proliferation; Cells, Cultured; Cyclooxygenase 2; Dinoprostone; Disease Models, Animal; Dose-Response Relationship, Drug; Down-Regulation; Endometriosis; Endometrium; Enzyme-Linked Immunosorbent Assay; Estradiol; Female; Gene Expression Regulation, Enzymologic; Humans; I-kappa B Proteins; Inflammation Mediators; Interleukin-8; Ki-67 Antigen; Mice; Mice, Inbred BALB C; Phosphorylation; Real-Time Polymerase Chain Reaction; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Sesquiterpenes; Signal Transduction; Stromal Cells; Transcription Factor RelA; Tumor Necrosis Factor-alpha

Endometriosis is characterized by repeated inflammatory changes and serious adhesions, inducing innate and adaptive immune responses within the abdominal cavity. To assess these immune responses, we evaluated the levels of expression of Toll-like receptors (TLR)-1, -2, -4, -5, and -9; nucleotide-binding oligomerization domains (NOD)-1 and -2; interleukins-1β, -6, -8, -10, and -12; interferon-γ; tumor necrosis factor-α; inducible nitric oxide synthase (iNOS) and endothelial NOS (eNOS); and immunoglobulins (Igs) in patients with endometriosis.. The levels of TLRs, NODs, cytokines, and NOS mRNAs in peritoneal effusions were assessed by real time reverse transcription-polymerase chain reaction; and IgG, IgA and IgM concentrations were measured by enzyme-linked immunosorbent assays (ELISA) in 40 patients with and 40 without endometriosis. Findings from the two groups were compared.. We observed expression of all pattern recognition receptors (PRRs), cytokines, and NOS mRNAs and Igs in the effusion fluid of patients with and without endometriosis. The levels of TLR-2 and -9; NOD-1 and -2; iNOS and eNOS mRNAs and CA 125 were significantly higher in the endometriosis than in the non-endometriosis group (p<0.05 each). Moreover, PRR, cytokine, and NOS expression showed significant correlations (p<0.05).. PRRs, cytokines, and NOS, which act cooperatively in the innate immune response, are closely associated with endometriosis. Increased expression of TLR-2, TLR -9, NOD-1, NOD-2, and NOS mRNA in peritoneal fluid may be associated with endometriosis.

Topics: Adult; Endometriosis; Enzyme-Linked Immunosorbent Assay; Female; Humans; In Vitro Techniques; Interleukin-10; Interleukin-12; Interleukin-1beta; Interleukin-6; Interleukin-8; Middle Aged; Nitric Oxide Synthase; Nitric Oxide Synthase Type II; Nitric Oxide Synthase Type III; Nod1 Signaling Adaptor Protein; Nod2 Signaling Adaptor Protein; Real-Time Polymerase Chain Reaction; Receptors, Pattern Recognition; Toll-Like Receptor 1; Toll-Like Receptor 2; Toll-Like Receptor 4; Toll-Like Receptor 9

Interleukin-22 (IL-22) is a member of the IL-10 cytokine family and plays critical roles in inflammation, immune surveillance, and tissue homeostasis. However, whether IL-22 regulates the growth of endometrial stromal cells (ESCs), and participates in the pathogenesis of endometriosis remain unclear. In this study, we found that the expression of IL-22 and it receptors (IL-22R1 and IL-10R2) in eutopic endometrium and ectopic lesion of women with endometriosis was higher than that from healthy control. Recombinant human IL-22 (rhIL-22) stimulated the proliferation of ESCs in a dosage-dependent manner. On the contrary, anti-human IL-22 neutralizing antibody inhibited the proliferation of ESCs in vitro. The stimulatory effect of IL-22 on the proliferation of ESCs could be reversed by inhibitor of STAT5, ERK1/2 or AKT signal pathway. However, blocking STAT3, JNK or P38 signal pathway had no these effects. By Enzyme-linked immunosorbent assay (ELISA) and flow cytometry assay, we demonstrated the rhIL-22 not only stimulate the secretion of CCL2 and IL-8, but also significantly up-regulate the expression of IL-8 receptor CXCR1 on ESCs. Meanwhile, STAT5, ERK1/2 and or AKT signal inhibitors could abrogate the increase of CCL2, IL-8 and CXCR1 levels induced by rhIL-22. However, rhIL-22 had not similar influence on CCL2 receptor CCR2. Our current results suggested that the higher level of IL-22 and it receptors in eutopic endometrium may stimulate the expression of CCL2, IL-8/CXCR1, and further promote the growth of ESCs possibly through activating STAT5, MAPK/ERK1/2 and or AKT signal pathways, which may be involved in the occurrence and development of endometriosis.

Topics: Cell Proliferation; Chemokine CCL2; Dose-Response Relationship, Drug; Endometriosis; Endometrium; Female; Humans; Interleukin-22; Interleukin-8; Interleukins; Receptors, Interleukin; Receptors, Interleukin-10; Signal Transduction; Stromal Cells

Nonmetastatic gene 23-H1 (NME1, also known as nm23-H1) is a wide-spectrum tumor metastasis suppressor gene that plays an important role in suppressing the proliferation, adhesion and invasion of endometrial stromal cells (ESCs). The present study is undertaken to explore the mechanism by which NME1 in ESCs from endometriosis modulates the angiogenesis and herein participates in the pathogenesis of endometriosis. The expression of NME1 in the primary ESCs from normal endometrium without endometriosis was higher than that from eutopic endometrium and ectopic lesion with endometriosis. Silencing NME1 stimulated the secretion of angiogenic factors interleukin-8 (IL-8) and vascular-endothelial growth factor (VEGF) of the eutopic ESCs from women with endometriosis, and these effects could be abrogated by MAPK/ERK1/2 or AKT inhibitor. In addition, the supernatant of NME1-silenced ESCs increased the expression of angiogenesis-relative molecules CD62E and CD105, and promoted angiogenesis of human umbilical vein endothelial cells (HUVECs). Anti-human IL-8 or VEGF neutralizing antibody reversed the effect on angiogenesis of HUVECs induced by NME1-silenced ESCs. Our current results suggest that the abnormal lower expression of NME1 in ESCs secrete more IL-8 and VEGF through activation of MAPK/ERK1/2 and AKT signal pathways, up-regulate the level of CD62E and CD105, and finally lead to numerous angiogenesis of vascular endothelial cells in the endometriotic milieu, which is beneficial to the origin and development of endometriosis.

Topics: Adult; Antigens, CD; E-Selectin; Endoglin; Endometriosis; Endometrium; Endothelial Cells; Female; Humans; Interleukin-8; Middle Aged; Neovascularization, Pathologic; NM23 Nucleoside Diphosphate Kinases; Receptors, Cell Surface; Stromal Cells; Up-Regulation; Vascular Endothelial Growth Factor A

We investigated the potential of gonadotropin-releasing hormone (GnRH) agonists and GnRH antagonists to inhibit cell proliferation in endometriotic and endometrial stromal cells.. Twenty patients with ovarian endometriomas and 18 patients with uterine fibromas were recruited. Endometriotic and endometrial stromal cells were obtained from the ovarian chocolate cyst linings and the eutopic endometria of premenopausal women with uterine fibromas, respectively.. GnRH agonist or antagonist treatment attenuated tumor necrosis factor (TNF)-α-induced cell proliferation in the endometrial stromal cells, whereas endometriotic stromal cells did not respond to treatment. The endometriotic stromal cells exhibited a decreased expression of the type I GnRH receptor compared with the endometrial stromal cells. GnRH agonists or antagonists did not repress TNF-α-induced IL-8 production in endometriotic stromal cells.. GnRH agonists and antagonists have similar effects in slowing the growth of endometrial stromal cells. Endometriotic stromal cells resist the antiproliferative effect of GnRH agonists and antagonists.

Topics: Adult; Blotting, Western; Buserelin; Cell Proliferation; Endometriosis; Endometrium; Enzyme-Linked Immunosorbent Assay; Female; Gene Expression; Gonadotropin-Releasing Hormone; Humans; Interleukin-8; Leiomyoma; Ovarian Diseases; Premenopause; Receptors, LHRH; Stromal Cells; Tumor Necrosis Factor-alpha; Uterine Neoplasms

Pelvic endometriosis is a chronic inflammatory disease with an immunological background. Yet there is paucity of contemporary research exploring both the angiogenic cytokines, leptin and IL-8 for a possible role in its pathophysiology.. To compare levels of both leptin and IL-8 in peritoneal fluid (PF) in women with endometriosis vs. fertile controls and correlate with disease stage, type and symptoms.. PF from 58 women with endometriosis and 28 women undergoing tubal ligation was collected at laparoscopy and leptin and IL-8 levels were measured using ELISA. Results showed significantly higher levels of both cytokines in women with endometriosis. Significantly higher leptin and IL-8 levels were demonstrated in patients with early peritoneal (ASRM stage I and II) and advancing disease (ASRM stage III and IV), respectively. Levels of leptin/IL-8 were significantly lower in patients with endometrioma (4.8 ng/mL/32 pg/mL) vs. implants (13.0 ng/mL/68 pg/mL). There was no correlation of infertility or chronic pelvic pain with these levels.. Both leptin and IL-8 levels are raised in PF of women with endometriosis reflecting inflammation and dysregulated immunomodulation. Higher levels of leptin were seen in early stages; IL-8 seems to stimulate the disease in a dose-dependent manner.

Topics: Adult; Ascitic Fluid; Case-Control Studies; Endometriosis; Female; Humans; Infertility, Female; Interleukin-8; Leptin; Ovarian Diseases; Pelvic Pain; Peritoneal Diseases

MicroRNAs have recently been identified as regulators that modulate target gene expression and are suggested to be involved in the development and progression of endometriosis. This study was undertaken to analyze the expression level of microRNA-199a (miR-199a) in paired ovarian endometrioma and eutopic endometrium from women with endometriosis, and to investigate the contribution of miR-199a to the invasive capability of endometrial stromal cells (ESCs). Cell adhesion, migration and Matrigel invasion assays were carried out to measure the invasiveness of ESCs. Bioinformatics prediction, reporter gene assay, PCR, western blotting and ELISA were performed to identify miR-199a targets and related signaling pathways. The results showed that the expression level of miR-199a was lower in the eutopic endometrium from women with endometriosis, and even lower in the paired ovarian endometrioma, compared with the expression in normal controls. Moreover, ectopic expression of miR-199a attenuated ESC adhesion, migration and invasiveness. MiR-199a targeted and inhibited IkappaB kinase beta (IKKβ) in ESCs. Accompanied by IKKβ reduction, miR-199a suppressed nuclear factor-kappa B (NF-κB) pathway activation and interleukin-8 (IL-8) production in ESCs. All these findings suggest that miR-199a, down-regulated in endometriosis, attenuates the invasive capability of ESCs in vitro partly through IKK/NF-κB pathway suppression and reduced IL-8 expression. In conclusion, miR-199a could be involved in the pathogenesis of endometriosis.

Topics: Adult; Blotting, Western; Cell Adhesion; Cell Movement; Cells, Cultured; Endometriosis; Endometrium; Enzyme-Linked Immunosorbent Assay; Female; Humans; I-kappa B Kinase; In Vitro Techniques; Interleukin-8; MicroRNAs; NF-kappa B; Real-Time Polymerase Chain Reaction; Signal Transduction; Stromal Cells; Young Adult

Curcumin, a naturally occurring polyphenolic compound from Curcuma longa, has long been used in folk medicine as an antiinflammatory remedy in Asian countries. Endometriosis is a chronic gynecological inflammatory disorder in which immune system deregulation may play a role in its initiation and progression. A number of mediators, including cell adhesion molecules such as intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1); proinflammatory cytokines such as tumour necrosis factor-α (TNF-α), interleukin-1 (IL-1), IL-6 and IL-8; and chemokines such as monocyte chemotactic protein-1 (MCP-1), play key roles in the pathogenesis of endometriosis. The aim of our study was to explore the effect of curcumin on the expression of these critical molecules in human ectopic endometriotic stromal cells isolated from women with endometriosis. Endometriotic stromal cells treated with curcumin showed marked suppression of TNF-α-induced mRNA expression of ICAM-1 and VCAM-1. Curcumin treatment also significantly decreased the TNF-α-induced cell surface and total protein expression of ICAM-1 and VCAM-1 in a dose-dependent manner. In addition, treatment of endometriotic stromal cells with curcumin markedly inhibited TNF-α-induced secretion of IL-6, IL-8 and MCP-1. Furthermore, curcumin inhibited the activation of transcription factor NF-κB, a key regulator of inflammation, in human endometriotic stromal cells. These findings suggest that curcumin may have potential therapeutic uses in the prevention and treatment of endometriosis.

Topics: Adult; Cells, Cultured; Chemokine CCL2; Curcumin; Dose-Response Relationship, Drug; Endometriosis; Female; Humans; Intercellular Adhesion Molecule-1; Interleukin-6; Interleukin-8; Middle Aged; NF-kappa B p50 Subunit; Stromal Cells; Tumor Necrosis Factor-alpha; Vascular Cell Adhesion Molecule-1

Activin A is an endometrial secretory product involved in inflammation and angiogenesis. The present study aimed to evaluate the effect of activin A and its antagonist follistatin on interleukin (IL)-6, IL-8, and vascular endothelial growth factor (VEGF) expression and release from cultured human endometrial stromal cells (HESCs) from women with and without endometriosis.. The HESCs were collected from women with endometriosis (n = 6) and controls (n = 6). Primary cultures were treated with activin A at different doses or activin A plus follistatin. The IL-6, IL-8, and VEGF messenger RNA expression was evaluated by real-time polymerase chain reaction and protein release was evaluated by enzyme-linked immunosorbent assay.. Unstimulated HESC from women with endometriosis secreted more IL-6 and IL-8 than controls. The addition of activin A increased IL-8 and VEGF secretion in HESC from controls and decreased IL-6 and IL-8 secretion in HESC from women with endometriosis. These effects were counteracted by follistatin.. Activin A regulates the expression and secretion of IL-8 and VEGF in cultured HESC, and this mechanism appears to be disrupted in eutopic endometrial cells from women affected by endometriosis.

Topics: Activins; Adult; Cells, Cultured; Endometriosis; Endometrium; Female; Follistatin; Gene Expression Regulation; Humans; Interleukin-6; Interleukin-8; RNA, Messenger; Stromal Cells; Vascular Endothelial Growth Factor A

To evaluate inflammatory/angiogenic cytokines-interleukin-1β (IL-1β), IL-6, IL-8, IL-12, interferon-γ (IFN-γ), tumor necrosis factor (TNF), and vascular endothelial growth factor A (VEGF-A)-in the peritoneal fluid of patients with endometriosis in relation to the occurrence and severity of pelvic adhesions and in control women without pelvic pathology.. Case-control study.. University research institution and hospital.. Sixty-five women with laparoscopically and histopathologically confirmed endometriosis, including 40 women with pelvic adhesions, and 37 control women without pelvic pathology.. Peritoneal fluid aspirated during routine diagnostic laparoscopic examination.. Cytokines evaluated in the peritoneal fluid via specific enzyme-linked immunosorbent assays.. Endometriosis and the revised American Fertility Society score of this disease were associated with statistically significantly increased levels of peritoneal IL-6 and IL-8 whereas the incidence and score of endometriosis-related pelvic adhesions were negatively associated with increased levels of VEGF-A. Notably, the concentration of VEGF-A predicted adhesion development and severity after adjustment for endometriosis severity. The adhesion score also correlated with increased levels of IL-6; however, after adjustment for endometriosis severity, the effect of this cytokine was no longer statistically significant.. Increased levels of VEGF-A may be associated with a decreased rate of pelvic adhesion formation in the course of endometriosis.

Topics: Ascitic Fluid; Case-Control Studies; Cytokines; Endometriosis; Female; Humans; Interferon-gamma; Interleukin-12; Interleukin-1beta; Interleukin-6; Interleukin-8; Peritonitis; Severity of Illness Index; Tissue Adhesions; Tumor Necrosis Factor-alpha; Vascular Endothelial Growth Factor A

Chemokine CXCL8 (also known as IL-8) has been identified as a potential regulator of endometrial stromal cells (ESCs), but it is unclear how CXCL8 regulates the survival of ESCs in the pathogenesis of endometriosis.. We assessed the secretion of CXCL8 by enzyme-linked immunosorbent assays and the expression of its receptors, CXCR1 and CXCR2, by in-cell Western assay and immunohistochemistry. The effects of CXCL8 on the activation or expression of various cell mediators were also investigated by in-cell Western assay. The effects of CXCL8 on the proliferation, growth and apoptosis of ESCs in vitro were assessed by BrdU assays, cell counts and annexin V labeling, respectively.. Secretion of CXCL8 and expression of CXCR1 in the eutopic ESCs from women with endometriosis were significantly higher than that in control ESCs, but the expression of CXCR2 showed no significant difference between these two cell types. CXCL8 stimulated proliferation and growth and reduced apoptosis of ESCs in an autocrine manner, and these effects were abolished by anti-human CXCL8 and CXCR1 neutralizing antibodies and by a PI3K/Akt inhibitor. Moreover, CXCL8 up-regulated the expression of the anti-apoptotic proteins, survivin and Bcl-2, inhibited the expression of the Phosphatase and tensin homolog (PTEN) and activated the phosphorylation of Akt.. This study suggests that CXCL8 and CXCR1 are involved in the pathogenesis of endometriosis by up-regulating proliferation and growth and restricting apoptosis in ESCs by activating the PTEN/Akt pathway and mediating the expression of survivin and Bcl-2.

Topics: Adult; Apoptosis; Bromodeoxyuridine; Cell Enlargement; Cell Proliferation; Endometriosis; Endometrium; Enzyme-Linked Immunosorbent Assay; Female; Humans; Immunohistochemistry; Inhibitor of Apoptosis Proteins; Interleukin-8; Middle Aged; Phosphorylation; Proto-Oncogene Proteins c-akt; Proto-Oncogene Proteins c-bcl-2; PTEN Phosphohydrolase; Receptors, Interleukin-8A; Survivin; Up-Regulation

Endometriosis, a disease of reproductive age women, is a major cause of infertility, menstrual disorders and pelvic pain. Little is known about its etiopathology, but chronic pelvic inflammation is a common feature in affected women. Beside symptomatic treatment of endometriosis-associated pain, only two main suboptimal therapeutic approaches (hormonal and invasive surgery) are generally recommended to patients and no specific targeted treatment is available. Our studies led to the detection of a marked increase in the expression of macrophage migration inhibitory factor (MIF) in the eutopic endometrium, the peripheral blood and the peritoneal fluid of women with endometriosis, and in early, vascularized and active endometriotic lesions. Herein, we developed a treatment model of endometriosis, where human endometrial tissue was first allowed to implant into the peritoneal cavity of nude mice, to assess in vivo the effect of a specific antagonist of MIF (ISO-1) on the progression of endometriosis and evaluate its efficacy as a potential therapeutic tool. Administration of ISO-1 led to a significant decline of the number, size and in situ dissemination of endometriotic lesions. We further showed that ISO-1 may act by significantly inhibiting cell adhesion, tissue remodeling, angiogenesis and inflammation as well as by altering the balance of pro- and anti-apoptotic factors. Actually, mice treatment with ISO-1 significantly reduced the expression of cell adhesion receptors αv and ß3 integrins (P<0.05), matrix metalloproteinases (MMP) 2 and 9 (P<0.05), vascular endothelial cell growth factor (VEGF) (P<0.01), interleukin 8 (IL8) (P<0.05), cyclooxygenease (COX)2 (P<0.001) and the anti-apoptotic protein Bcl2 (P<0.01), but significantly induced the expression of Bax (P<0.05), a potent pro-apoptotic protein. These data provide evidence that specific inhibition of MIF alters endometriotic tissue growth and progression in vivo and may represent a promising potential therapeutic avenue.

Topics: Animals; Cell Adhesion; Cyclooxygenase 2; DNA Primers; Endometriosis; Female; Gene Expression Regulation; Histological Techniques; Humans; Integrin alphaV; Integrin beta3; Interleukin-8; Intramolecular Oxidoreductases; Isoxazoles; Macrophage Migration-Inhibitory Factors; Matrix Metalloproteinase 2; Matrix Metalloproteinase 9; Mice; Mice, Nude; Neovascularization, Pathologic; Real-Time Polymerase Chain Reaction; Statistics, Nonparametric; Vascular Endothelial Growth Factor A

We analyzed selected well-known and less well-known serum markers that have been proposed for diagnosis and severity assessment of endometriosis, in a case-control study.. This prospective study was carried out in a Clinical Department of Gynecology in Iasi, Romania. Study participants included endometriosis patients, and controls in whom laparoscopy had excluded endometriosis. Each case and control was investigated for serum levels of CA125, TNF, IL-1, IL-6 and IL-8. The data were correlated with clinical symptoms and revised American Fertility Society (rAFS) score and stage, and interpreted by Mann-Whitney U-test and ANOVA regression analysis.. Over the course of 1 year, 24 cases of endometriosis and 24 controls of matched age were selected. The rAFS stages were: stage I, 12.5%; stage II, 16.7%; stage III, 58.3%; and stage IV, 12.5%. CA125 levels were over the cut-off of 35 IU/l in 54% of patients (versus 8% of controls), averaging 67.5 (CI95: ±17.5). The sensitivity and specificity were 54% and 91%, respectively, with a p value of <0.001 (statistically significant). For IL-6, 71% of cases and 87% of controls were above the cut-off of 2 pg/ml, with an average of 11.83 ± 7. The sensitivity and specificity were 71% and 12%, respectively, but the difference was not statistically significant, p = 0.071. Other tested serum markers had no discrimination value. A correlation with severity of endometriosis was seen for CA125 (p = 0.03) but not for IL-6, by ANOVA.. CA125 correlated with endometriosis screening and severity, indicating its superiority as a marker for further, larger studies.

Topics: Adult; Biomarkers; CA-125 Antigen; Case-Control Studies; Endometriosis; Female; Humans; Interleukin-1; Interleukin-6; Interleukin-8; Prognosis; Prospective Studies; Sensitivity and Specificity

To examine the effects of interleukin (IL)-17F on the secretion of IL-8 and the gene expression of cyclooxygenase 2 (COX2) in endometriotic stromal cells.. In vitro experimental study using human samples.. University hospital.. Endometriotic tissues were obtained from women with ovarian endometriomas undergoing laparoscopic surgery.. Endometriotic stromal cells (ESCs) were cultured with IL-17F.. Concentrations of IL-8 were measured by a specific ELISA, and messenger RNA levels of IL-8 and COX2 were measured by real-time reverse transcription-polymerase chain reaction (PCR).. IL-17F increased the secretion of IL-8 from ESCs, and the effect was inhibited by antibodies for IL-17 receptor A and IL-17 receptor C. Tumor necrosis factor α (TNF-α) synergistically enhanced IL-17F-induced increase in IL-8 secretion from ESCs. The IL-17F increased the gene expression of IL-8 and COX2 in ESCs.. These findings suggest that IL-17F may stimulate the development of endometriosis by up-regulation of IL-8 and COX2.

Topics: Cells, Cultured; Cyclooxygenase 2; Endometriosis; Endometrium; Female; Gene Expression Regulation, Enzymologic; Humans; Interleukin-17; Interleukin-8

Inflammatory mediators, including chemokines, may play crucial roles in the development of endometriosis. Therefore, we investigated the expression and localization of CXCL16 and its receptor, CXCR6, in ovarian endometriotic tissues. We also examined whether CXCL16 induces IL-8 production in endometriotic stromal cells.. We performed immunohistochemical and Western blotting analyses of in vivo and in vitro samples. IL-8 production was assayed using an ELISA.. Both CXCL16 and CXCR6 were expressed by endometriotic epithelial cells and stromal cells, but not normal ovarian stroma. A Western blotting analysis using primary cultured endometriotic stromal cells showed a constant expression of CXCL16 and CXCR6 in the proliferative phase, secretory phase and during gonadotropin-releasing hormone agonist therapy. CXCL16 induced IL-8 production in several endometriotic stromal cells in vitro.. CXCL16 and CXCR6 might be involved in the pathophysiology of endometriosis through regulation of the inflammatory response.

Topics: Adult; Blotting, Western; Chemokine CXCL16; Chemokines, CXC; Endometriosis; Enzyme-Linked Immunosorbent Assay; Female; Gene Expression; Humans; Immunohistochemistry; Interleukin-8; Middle Aged; Ovarian Cysts; Receptors, Chemokine; Receptors, CXCR6; Receptors, Scavenger; Receptors, Virus; Stromal Cells

To explore the effect and mechanism of Jiawei Foshou San on the accretion ectopic endometrium of rats.. Endometriosis model was established by surgical implant of endometrial tissue which belongs to its body in rats. Jiawei Foshou San was administrated to the model rats. Twenty-eight days later, the length of ectopic endometrium was measured by vernier caliper, the spleen was weighed by electronic balance, the content of tumor necrosis factor (TNF)-alpha in blood serum and peritoneal fluid was detected by ELISA test, the expression of interleukin (IL)-8 in ectopic endometrium was detected by immunohistochemical method, the expression of NF-kappaB p65 protein and inhibitory KBalpha (IkappaBalpha) protein in ectopic endometrium were analyzed by western blot.. Jiawei Foshou San 0.045, 0.09, 0.18 g x kg(-1) group reduced the volume of ectopic endometrium. Jiawei Foshou San 0.18 g x kg(-1) group raised the spleen exponent of model rats. Jiawei Foshou San 0.09, 0.18 g x kg(-1) group decreased the content of TNF-alpha in blood serum and peritoneal fluid, and the content of IL-8 in ectopic endometrium was also decreased. Jiawei Foshou San can decrease the expression of NF-KB p65 and increase the expression of IkappaBalpha in ectopic endometrium.. Jiawei Foshou San can inhibit the accretion of endometriosis implants of rats, and its mechanism might be associated with improving the environment of body.

Topics: Animals; Drugs, Chinese Herbal; Endometriosis; Endometrium; Female; Humans; Interleukin-8; Male; Rats; Rats, Sprague-Dawley; Tumor Necrosis Factor-alpha

To determine whether interleukin-8 (IL-8) serum levels are correlated with pelvic pain in patients with ovarian endometriomas.. Prospective study.. Tertiary-care university hospital.. Interleukin-8 serum levels were prospectively analyzed in 51 patients (group A, asymptomatic patients or patients with mild dysmenorrhea; group B, severe dysmenorrhea and/or chronic pelvic pain and/or dyspareunia) who underwent surgery for cystic ovarian endometriosis to asses whether a correlation exists between IL-8 serum levels and pelvic pain.. Interleukin-8 serum levels determination.. Interleukin-8 serum levels and pelvic pain.. From 56 patients, five cases were ultimately excluded because the histologic diagnosis was not cystic ovarian endometriosis (2 teratomas and 3 haemorragic cysts). The mean (+/-SD) IL-8 serum levels in group A were 6.41 +/- 12.17 pg/mL and in group B were 6.52 +/- 8.73 pg/mL.. Pain symptoms in ovarian endometriosis is not correlated with IL-8 serum levels.

Topics: Adult; Biomarkers; Chronic Disease; Dysmenorrhea; Endometriosis; Female; Humans; Interleukin-8; Ovarian Cysts; Pelvic Pain; Prospective Studies

Lack of a non-invasive diagnostic test contributes to the long delay between onset of symptoms and diagnosis of endometriosis. The aim of this study was to evaluate the combined performance of six potential plasma biomarkers in the diagnosis of endometriosis.. This case-control study was conducted in 294 infertile women, consisting of 93 women with a normal pelvis and 201 women with endometriosis. We measured plasma concentrations of interleukin (IL)-6, IL-8, tumour necrosis factor-alpha, high-sensitivity C-reactive protein (hsCRP), and cancer antigens CA-125 and CA-19-9. Analyses were done using the Kruskal-Wallis test, Mann-Whitney test, receiver operator characteristic, stepwise logistic regression and least squares support vector machines (LSSVM).. Plasma levels of IL-6, IL-8 and CA-125 were increased in all women with endometriosis and in those with minimal-mild endometriosis, compared with controls. In women with moderate-severe endometriosis, plasma levels of IL-6, IL-8 and CA-125, but also of hsCRP, were significantly higher than in controls. Using stepwise logistic regression, moderate-severe endometriosis was diagnosed with a sensitivity of 100% (specificity 84%) and minimal-mild endometriosis was detected with a sensitivity of 87% (specificity 71%) during the secretory phase. Using LSSVM analysis, minimal-mild endometriosis was diagnosed with a sensitivity of 94% (specificity 61%) during the secretory phase and with a sensitivity of 92% (specificity 63%) during the menstrual phase.. Advanced statistical analysis of a panel of six selected plasma biomarkers on samples obtained during the secretory phase or during menstruation allows the diagnosis of both minimal-mild and moderate-severe endometriosis with high sensitivity and clinically acceptable specificity.

Topics: Biomarkers; C-Reactive Protein; CA-125 Antigen; CA-19-9 Antigen; Case-Control Studies; Endometriosis; Female; Humans; Interleukin-6; Interleukin-8; Logistic Models; Tumor Necrosis Factor-alpha

Evidence suggests that eutopic endometrium from women with endometriosis (US-E) has intrinsic functional anomalies compared with women without endometriosis (US-C). We hypothesized that differences in endometrial haptoglobin (eHp) mRNA and protein levels exist between eutopic endometrium from US-E and US-C and that inflammatory mediators may be involved.. Endometrial stromal cells and tissue explants from US-E (n = 18) and US-C (n = 18) were cultured (24 h/48 h for cells/explants) with interleukin (IL)-1alpha, -1beta, -6, -8 or tumor necrosis factor-alpha (TNF-alpha) at 0-100 ng/ml. eHp protein in media and mRNA levels were quantified by enzyme-linked immunosorbent assay and quantitative PCR.. In eutopic endometrial stromal cells from US-E, IL-1beta, IL-6 and TNF-alpha (10 ng/ml) increased eHp mRNA levels (P = 0.002, P < 0.001 and P < 0.001, respectively) and eHp protein (P = 0.023, 0.031 and 0.006, respectively) versus control. In endometrial tissues from US-E, IL-1beta, IL-6 and TNF-alpha increased eHp mRNA (P < 0.001, P = 0.017 and P < 0.001, respectively) and eHp protein (P < 0.001, P = 0.007 and 0.039, respectively) versus control. IL-1alpha and IL-8 had small or no effects on isolated endometrial cells or tissues. In US-C, IL-1beta, IL-8 and TNF-alpha each reduced eHp mRNA in endometrial stromal cells (all P < 0.001) versus control; IL-1alpha and IL-6 had no effect. eHp mRNA increased in endometrial tissues from US-C in response to IL-1beta (P = 0.008), IL-6 (P = 0.015) and TNF-alpha (P = 0.031) versus control; IL-1alpha or IL-8 had no effect.. Endometrium from US-E differentially responds to specific inflammatory cytokines by production of eHp. We propose that up-regulation of endometrial eHp by inflammatory mediators disrupts normal endometrial function and may facilitate the pathogenesis of endometriosis.

Topics: Cytokines; Endometriosis; Endometrium; Female; Haptoglobins; Humans; In Vitro Techniques; Inflammation Mediators; Interleukin-1alpha; Interleukin-1beta; Interleukin-6; Interleukin-8; RNA, Messenger; Stromal Cells; Tumor Necrosis Factor-alpha; Up-Regulation

An active angiogenesis is required for ectopic endometrial tissue growth. Our previous studies led to the identification of macrophage migration inhibitory factor (MIF), which is markedly elevated in active, vascularized, and early-stage endometriotic lesions, as a potent mitogenic factor for endothelial cells.. Our objective was to study the mechanisms by which MIF may stimulate angiogenesis in ectopic endometrial implantation sites.. Primary cultures of ectopic endometrial cells were exposed to MIF, and the release of major angiogenic factors with targeted disruption of MIF signaling pathways was assessed.. Patients were women found to have endometriosis during laparoscopy.. The study was conducted at a hospital and reproduction research laboratory.. Biopsies were removed from endometriotic lesions.. Vascular endothelial cell growth factor (VEGF), IL-8, and monocyte chemotactic protein-1 (MCP-1) mRNA and protein levels and expression and small interfering RNA silencing of MIF CD74/CD44 receptor complex and phosphorylation of ERK and p38 MAPKs were evaluated.. MIF markedly up-regulated VEGF, IL-8, and MCP-1 expression in endometriotic cells. Such an effect was abolished by (S,R)-3-(4-hydroxyphenyl)-4,5-dihydro-5-isoxazole acetic acid methyl ester (ISO-1), a specific inhibitor of MIF, and significantly down-regulated after specific small interfering RNA silencing of CD44 or CD74. MIF treatment strongly activated ERK and p38 MAPKs, and specific inhibitors of both pathways completely blocked basal and MIF-induced VEGF, IL-8, and MCP-1 synthesis.. These results show for the first time that MIF exerts a potent indirect angiogenic effect by interacting with ectopic endometrial cells and inducing the secretion of major angiogenic factors via CD44, CD74, and MAPK signaling pathways and provide evidence for a possible new mechanism underlying endometriosis development and pathophysiology.

Topics: Angiogenesis Inducing Agents; Antigens, Differentiation, B-Lymphocyte; Biopsy; Chemokine CCL2; DNA Primers; Endometriosis; Endometrium; Female; Gene Silencing; Histocompatibility Antigens Class II; Humans; Hyaluronan Receptors; Interleukin-8; Macrophage Migration-Inhibitory Factors; Mitogen-Activated Protein Kinase Kinases; Neovascularization, Physiologic; p38 Mitogen-Activated Protein Kinases; Phenotype; Reverse Transcriptase Polymerase Chain Reaction; RNA Interference; RNA, Messenger; RNA, Small Interfering; Signal Transduction; Vascular Endothelial Growth Factor A

We previously reported that lipopolysaccharide (LPS)-promoted endometriotic stromal cell (ESC) proliferation by inducing TNFalpha production. The aim of this study was to investigate the efficacy of TNFalpha gene silencing on LPS-treated ESCs.. Endometriotic stromal cells (ESCs) and endometrial stromal cells (ESCs) (EMSCs) were obtained from ovarian chocolate cysts and uterine myoma, respectively. Using PCR array, LPS-induced gene expression profiling after transfection of TNFalpha siRNA into ESCs was performed. Down-regulated genes by TNFalpha silencing were examined using real-time RT-PCR. Effect of TNFalpha silencing was examined using ELISA and BrdU incorporation, respectively.. In PCR array, TNFalpha silencing in ESCs repressed LPS-induced expression of cIAP2 and IL-8, NFkappaB pathway responsive genes. After adding LPS, the levels of cIAP2 and IL-8 expression in ESCs were higher compared with those in EMSCs. TNFalpha silencing attenuated the LPS-induced ESC proliferation.. Tumor necrosis factor alpha may be involved in cell proliferation of endometriotic tissues.

Topics: Apoptosis; Baculoviral IAP Repeat-Containing 3 Protein; Cell Proliferation; Cells, Cultured; Endometriosis; Endometrium; Female; Gene Expression Profiling; Gene Silencing; Humans; Inhibitor of Apoptosis Proteins; Interleukin-8; Leiomyoma; Lipopolysaccharides; NF-kappa B; Ovarian Cysts; Ovarian Diseases; RNA, Small Interfering; Signal Transduction; Stromal Cells; Tumor Necrosis Factor-alpha; Ubiquitin-Protein Ligases; Uterine Neoplasms

Endometriosis causes pelvic pain and infertility in women of reproductive age. We explored TNFalpha-induced specific signaling pathways and gene expressions in endometriotic stromal cells (ESCs). Based on the data of the pathway specific cDNA array, we analyzed the role of TAK1, which is believed to work as a common mediator for NF-kappaB and MAPK pathways. Using the NF-kappaB pathway array, we found that TNFalpha upregulated ICAM-3, IL-6, IL-8, TAK1, JNK2, RelA, and TLR4 expressions. TNFalpha augmented the phosphorylation of TAK1. By transfection of TAK1 siRNA, TNFalpha-induced phosphorylation of IkappaBalpha, JNK1/2, and p38MAPK, as well as IL-6 or IL-8 expression, were repressed. TAK1 silencing in TNFalpha-pretreated ESCs caused a decrease in the proportion of cells in S-phase, and reduced TNFalpha-promoted BrdU incorporation. We provide the first evidence that TNFalpha and its downstream TAK1, which are key mediators for NF-kappaB and MAPK pathways, may be involved in the pathogenesis of endometriosis.

Topics: Cell Proliferation; Cytokines; DNA, Complementary; Endometriosis; Enzyme Activation; Female; Gene Expression Regulation; Gene Silencing; Humans; Interleukin-6; Interleukin-8; JNK Mitogen-Activated Protein Kinases; MAP Kinase Kinase Kinases; NF-kappa B; Oligonucleotide Array Sequence Analysis; p38 Mitogen-Activated Protein Kinases; Phosphorylation; Stromal Cells; Tumor Necrosis Factor-alpha

Intraperitoneal immuno-inflammatory changes may be associated with the pathogenesis of endometriosis. We evaluated the effects of peritoneal fluid obtained from patients with endometriosis (ePF) on the release of interferon-gamma-induced protein-10 (IP-10/CXCL10) and interleukin-8 (IL-8/CXCL8) by neutrophils, CD4(+) T cells, and monocytes.. Neutrophils, CD4(+) T cells, and monocytes were cultured with ePF and the chemokine levels in the supernatants were then measured using enzyme-linked immunosorbent assay.. The addition of ePF to cultures of CD4(+) T cells led to a significant increase in the release of IP-10 when compared with control PF without endometriosis (cPF). There was a positive correlation between the levels of IL-8 and IP-10 in ePF (R = 0.89, P = 0.041), but not between the levels of IP-10 and IL-8 released by neutrophils, CD4(+) T cells, and monocytes. The levels of IP-10 in ePF were positively correlated with the release of IP-10 by ePF-treated neutrophils (R = 0.89, P < 0.001), CD4(+) T cells (R = 0.93, P < 0.001), and monocytes (R = 0.70, P = 0.01). Moreover, the addition of ePF significantly enhanced the interferon-gamma-induced release of IP-10 by nuetrophils and CD4(+) T cells.. These findings suggest that neutrophils and T cells release differential levels of IP-10 and IL-8 in response to stimulation with ePF, and that these cells are a major source of IP-10 in the PF of endometriosis patients.

Topics: Adult; Ascitic Fluid; CD4-Positive T-Lymphocytes; Cells, Cultured; Chemokine CXCL10; Coculture Techniques; Endometriosis; Female; Humans; Interferon-gamma; Interleukin-8; Neutrophils

To investigate the expression and localization of interleukin-8 (IL-8) and monocyte chemotactic protein 1 (MCP-1) in women with and without endometriosis.. Comparative immunohistochemical study.. Academic medical center.. Ectopic (n = 24) and homologous eutopic endometrium (n = 24) from women with endometriosis and endometrium from women without endometriosis (n = 27) were used for immunohistochemical analysis of IL-8 and MCP-1.. Tissue sections were immunostained with antihuman IL-8 and MCP-1 antibodies.. Microscopic evaluation to assess the presence and localization of IL-8 and MCP-1 throughout the menstrual cycle in both eutopic endometrial and endometriotic tissues of women with endometriosis and comparison with normal endometrium.. In normal endometrium, secretory phase samples expressed higher levels of epithelial IL-8 than in proliferative phase samples. Epithelial MCP-1 expression was similar in both proliferative and secretory phases. Proliferative phase samples showed higher epithelial IL-8 and MCP-1 expressions in eutopic endometrium of women with endometriosis compared with that of normal women. Immunoreactivities of both chemokines were significantly increased in the epithelial cells of ectopic endometrial tissues compared with those of normal endometrium.. These findings suggest that IL-8 and MCP-1 may be involved in the pathogenesis of endometriosis.

Topics: Adult; Case-Control Studies; Chemokine CCL2; Endometriosis; Endometrium; Epithelial Cells; Female; Humans; Immunohistochemistry; Interleukin-8; Middle Aged; Retrospective Studies; Stromal Cells

To determine whether serum interleukin (IL)-8 concentration can be measured in patients with ovarian endometrioma and whether this measurement is a useful tool in diagnosing this disease.. A controlled clinical study and an in vitro study.. Department of Obstetrics and Gynecology, Tottori University, Japan.. Seventy patients with ovarian endometrioma and 21 patients with benign ovarian cyst.. Laparoscopic surgery or laparotomy for ovarian endometriomas or benign ovarian cyst was performed. Preoperative blood samples were obtained. Endometriotic stromal cells obtained from nine patients with endometrioma were cultured.. Interleukin-8 concentration in the serum or supernatant of the cell culture was measured with use of ELISA.. The serum concentration of IL-8 in patients with endometrioma was significantly higher than in patients with benign ovarian cyst. The serum IL-8 threshold (25 pg/mL) had a higher sensitivity (71.4%) for diagnosing ovarian endometrioma than did serum CA-125 level. The increased rates of IL-8 concentration in the culture supernatants after adding tumor necrosis factor alpha were significantly higher in patients whose serum IL-8 levels were >or=25 pg/mL than in those with levels <25 pg/mL.. Measuring of serum IL-8 concentration may be a valuable tool in diagnosing endometriosis.

Topics: Adult; Endometriosis; Female; Humans; Interleukin-8; Middle Aged; Ovarian Diseases; Reproducibility of Results; Sensitivity and Specificity

The pathogenesis of endometriosis is related to functional changes in CD3+ and CD14+ cells observed both at the local and systemic level. Here we investigated whether, and if so, how the body compartment influences cytokine expression in stimulated peritoneal and peripheral CD3+ and CD14+ cells of women with endometriosis.. Isolated peripheral blood (PB) and peritoneal fluid (PF) mononuclear cells from women with endometriosis were cultured under non-adherent conditions and stimulated with PMA and ionomycin for 6h to induce intracellular cytokine synthesis of TNF-alpha, IFN-gamma, and IL-8 by CD3+ cells or with LPS for 9h to produce TNF-alpha, IL-6, IL-10, MCP-1, and IL-8 by CD14+ cells.. The percentages of positive CD3+ cells stained for TNF-alpha and IFN-gamma were significantly higher and those stained for IL-8 were significantly lower in PF compared with PB, this being independent of the stage of endometriosis. In contrast, the percentages of CD14+ cells producing TNF-alpha, IL-6, IL-10, MCP-1, and IL-8 were significantly higher in PB than PF of women with endometriosis.. Monocytes/macrophages and lymphocytes derived from the peripheral and peritoneal compartments of women with endometriosis differentially respond to stimulated cytokine synthesis induction. However, it is difficult to state whether the observed phenomenon is more related to body compartment influence per se or to the presence of endometriosis.

Topics: Adult; Ascitic Fluid; CD3 Complex; Chemokine CCL2; Cytokines; Endometriosis; Female; Flow Cytometry; Humans; Interferon-gamma; Interleukin-10; Interleukin-6; Interleukin-8; Leukocytes, Mononuclear; Lipopolysaccharide Receptors; Lymphocytes; Macrophages; Tumor Necrosis Factor-alpha

To determine the role of interleukin-1 receptor type II (IL-1RII) in the pathogenesis of endometriosis.. Cultures of endometrial cells exposed to soluble IL-1RII or the recombinant adenovirus of IL-1RII (rAd-RII).. Gynecology clinic and human reproduction research laboratory.. Women with endometriosis undergoing hysterectomy.. Cell culture media were collected 12 hours after addition of soluble IL-1RII or infection of rAd-RII.. The levels of IL-6 and IL-8 in the culture media were measured via enzyme-linked immunoabsorbent assay. Furthermore, proteins of the cells were collected for two-dimensional electrophoresis and the differential protein expression was identified by the matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry.. Addition of soluble IL-1RII (2.0 microg/mL) significantly inhibited IL-1 beta-induced IL-6 and IL-8 secretion by endometrial cells in vitro. Infection of endometrial cells with rAd-RII significantly decreased IL-1 beta-induced IL-8 secretion, compared with the PBS and rAd-LacZ controls but had no significant effect on IL-6 secretion. Proteins of the infected cells were collected for two-dimensional electrophoresis, and intensities of 62 spots were significantly increased or decreased when compared with those in the PBS group. Thirty-four proteins were identified by MALDI-TOF mass spectrometry. The majority of the identified proteins are related to cellular metabolism and proliferation.. These results suggest that IL-1RII can neutralize IL-1 beta and counteract its effect on endometrial stromal cells, and may provide a new clinical strategy for the treatment of endometriosis.

Topics: Adenoviridae; Adult; Cells, Cultured; Electrophoresis, Gel, Two-Dimensional; Endometriosis; Enzyme-Linked Immunosorbent Assay; Female; Genetic Vectors; Humans; Interleukin-1beta; Interleukin-6; Interleukin-8; Middle Aged; Proteins; Proteomics; Receptors, Interleukin-1 Type II; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Stromal Cells; Time Factors; Transfection

To evaluate the influence of peroxisome proliferator-activated receptor-gamma (PPAR gamma) ligand (pioglitazone) on tumor necrosis factor-alpha (TNF-alpha)-induced interleukin-8 (IL-8) expression in endometriotic stromal cells (ESCs) and on proliferation of ESCs.. Prospective study.. Department of Obstetrics and Gynecology, Tottori University Hospital, Yonago, Japan.. Twenty-seven patients who underwent laparoscopic surgery.. The ESCs were obtained from the chocolate cyst linings of ovaries.. The expression of PPAR gamma gene and protein was determined by reverse transcriptase-polymerase chain reaction (RT-PCR) and immunocytochemistry. We determined the effect of pioglitazone on the production of TNF-alpha-induced IL-8 protein in culture supernatant of ESCs using ELISA. The effect of pioglitazone on TNF-alpha-induced proliferation of ESCs was evaluated by 5-bromo-2'-deoxyuridine proliferation assay. The activation of nuclear factor (NF)-kappaB in ESCs was evaluated by Western blot analyses and NF-kappaB transcription factor assays.. Immunocytochemistry and RT-PCR revealed the expression of PPAR gamma gene and protein in ESCs. The PPAR gamma protein was predominantly located in the cell nucleus. Measurement of IL-8 protein by ELISA showed that adding TNF-alpha (100 pg/mL) significantly increased IL-8 protein. Treating ESCs with 0.1-10 microM of pioglitazone significantly reduced the TNF-alpha-induced IL-8 production. The presence of 0.1-10 microM of pioglitazone significantly suppressed growth of ESCs. The TNF-alpha increased the expression of phosphorylation of inhibitor kappaB (I kappaB). Adding pioglitazone (10 microM) did not influence the expression of phosphorylated inhibitor kappaB (I kappaB). The TNF-alpha markedly increased the intranuclear concentration of p65, and adding pioglitazone (10 microM) significantly reduced the concentration of p65.. The present study demonstrates for the first time that PPAR gamma is expressed in ESCs, and that pioglitazone reduced IL-8 secretion and the proliferation of ESCs. The PPAR gamma ligand may be an attractive therapeutic agent for endometriosis.

Topics: Cell Nucleus; Cell Proliferation; Cells, Cultured; Endometriosis; Female; Gene Expression; Humans; I-kappa B Proteins; Interleukin-8; Ligands; Pioglitazone; PPAR gamma; Stromal Cells; Thiazolidinediones; Transcription Factor RelA; Tumor Necrosis Factor-alpha

To test the ability of a group of serum cytokines, either individually or in combination, to serve as biomarkers for the nonsurgical diagnosis of endometriosis.. Subjects were allocated to two groups according to their laparoscopic diagnosis. The first group consisted of patients with endometriosis and the second group was made up of infertile women with no pelvic pathology (controls). Blood samples were collected preoperatively and stored. Cytokines were measured in the serum of all participants using the Bio-Plex Protein Array System. Nonparametric statistics and the Mann-Whitney test were used to compare groups. Subjects were seen at the Gynecologic endoscopy unit.. Three cytokines were significantly higher in the serum of subjects with endometriosis than in the control group: interleukin-6 (IL-6) [4.41 pg/ml (range: 1.47-15.01) versus 0.97 pg/ml (range: 0.29-2.98), respectively; p<0.001], monocyte chemotactic protein-1 (MCP-1) [37.91 pg/ml (range: 24.54-94.74) versus 22.13 pg/ml (range: 13.85-39.45), respectively; p<0.001], and interferon-gamma (INF-gamma) [19.01 pg/ml (range: 1.19-73.52) versus 0.30 pg/ml (range: 0.00-13.05), respectively; p<0.001]. There was no statistically significant difference between subjects with endometriosis and controls in the serum concentration of vascular endothelial growth factor (VEGF), tumor necrosis factor-alpha (TNF-alpha), or granulocyte macrophage colony stimulating factor (GM-CSF). Interleukin-2 (IL-2), interleukin-8 (IL-8), and interleukin-15 (IL-15) were undetectable in the serum of both groups. None of the measured cytokines showed significant correlation with the cycle phase or stage of endometriosis. In a multivariate analysis, serum interleukin-6 provided a sensitivity of 71% and a specificity of 66% to discriminate between endometriosis patients and controls at a cutoff point of 1.9 pg/ml. Adding monocyte chemotactic protein-1 and interferon-gamma to interleukin-6 did not increase the discriminative ability over that achieved by measuring serum interleukin-6 alone.. Serum of subjects with endometriosis contains significantly higher levels of interleukin-6, monocyte chemotactic protein-1, and interferon-gamma than control women. Serum interleukin-6 measurements discriminate between women with endometriosis and controls. Interleukin-6 provides a promising serum marker for the nonsurgical prediction of endometriosis.

Topics: Adult; Biomarkers; Case-Control Studies; Chemokine CCL2; Endometriosis; Female; Granulocyte-Macrophage Colony-Stimulating Factor; Humans; Interferon-gamma; Interleukin-15; Interleukin-2; Interleukin-6; Interleukin-8; Multivariate Analysis; Predictive Value of Tests; Sensitivity and Specificity; Tumor Necrosis Factor-alpha; Vascular Endothelial Growth Factor A

To analyze the constitutive production of epithelial neutrophil activating peptide 78 (ENA-78) and interleukin-8 (IL-8) by epithelial cells and the response of these cells to cytokine stimulation.. In vitro study using eutopic endometrial tissue.. University hospital.. Cycling women undergoing laparoscopy for reasons of infertility or unexplained abdominal pain.. Isolation of epithelial and stromal cells from endometrium, immunocytochemical characterization and separate culture of these cells in presence of IL-1, tumor necrosis factor alpha (TNF-alpha), and interferon-gamma.. Quantitation of IL-8 and ENA-78 released into the medium by ELISA. Polymerase chain reaction was used to demonstrate the presence of ENA-78 in the cell lysate.. High purity of the endometrial epithelial cell preparation before culture was demonstrated by the lack of immunocytochemical staining for CD10. Stromal cell preparations were CD10 positive and cytokeratin negative. Stromal cells produced ENA-78 and IL-8 under cytokine stimulation, and epithelial cells were found not only to produce these markers in the absence of cytokine stimulation, but also to increase this output in the presence of IL-1beta or of TNF-alpha plus interferon-gamma.. This response may be an important angiogenic step in the early stages in the pathogenesis of endometriosis.

Topics: Antibodies, Monoclonal; Cells, Cultured; Chemokine CXCL5; Cytokines; Endometriosis; Endometrium; Epithelial Cells; Female; Humans; Interferon-gamma; Interleukin-1beta; Interleukin-8; Neprilysin; RNA, Messenger; Stromal Cells; Tumor Necrosis Factor-alpha

IL-17A is secreted from Th17 cells, a discovery leading to revision of the mechanism underlying the role of Th1/Th2 in the immune response. Strong evidence suggests that immune responses associated with inflammation are involved in the pathogenesis of endometriosis. In the present study, we first demonstrated that the presence of Th17 cells in peritoneal fluid of endometriotic women by flow cytometric analysis and IL-17A-positive cells in endometriotic tissues by immunohistochemistry. To investigate the role of IL-17A in the development of endometriosis, we then studied the effect of IL-17A on IL-8 production, cyclooxygensase-2 expression, and cell proliferation of cultured endometriotic stromal cells (ESCs). IL-17A enhanced IL-8 secretion from ESCs in a dose-dependent manner. The IL-17A-induced secretion of IL-8 from ESCs was suppressed by anti-IL-17 receptor A antibodies or inhibitors of p38 MAPK, p42/44 MAPK, and stress-activated protein kinase/c-Jun N-terminal kinase. Addition of TNFalpha synergistically increased IL-17A-induced IL-8 secretion from ESCs. IL-17A also enhanced the expression of cyclooxygensase-2 mRNA and proliferation of ESCs. IL-17A may play a role in the development of endometriosis by stimulating inflammatory responses and proliferation of ESCs.

Topics: Antibodies, Anti-Idiotypic; Cell Proliferation; Cells, Cultured; Cyclooxygenase 2; Endometriosis; Endometrium; Female; Humans; Interleukin-17; Interleukin-8; Mitogen-Activated Protein Kinase Kinases; Phosphorylation; RNA, Messenger; Stromal Cells; T-Lymphocytes, Regulatory; Tumor Necrosis Factor-alpha

Tumor necrosis factor (TNF)-alpha is central to the endometriotic disease process. TNF-alpha receptor signaling regulates epithelial cell secretion of inflammation and invasion mediators. Because epithelial cells are a disease-inducing component of the endometriotic lesion, we explored the response of 12Z immortalized human epithelial endometriotic cells to TNF-alpha. This report reveals the impact of disruption of established TNF-alpha-induced signaling cascades on the expression of biomarkers of inflammation and epithelial-mesenchymal transition (EMT) from endometriotic epithelial cells. Note that we show the molecular potential of soluble TNF-R1 [TNF binding protein (TBP)] and a panel of small molecule kinase inhibitors to block endometriotic gene expression directly. The TNF-alpha receptor is demonstrated to signal through IkappaB kinase complex (IKK) 2 > IkappaB > nuclear factor kappaB, extracellular signal-regulated kinase > mitogen-activated protein kinase kinase (MEK), p38, and phosphatidylinositol 3-kinase (PI3K) > Akt1/2. TNF-alpha induces the expression of transcripts for inflammatory mediators interleukin (IL)-6, IL-8, regulated on activation normal T cell expressed and secreted, TNF-alpha, granulocyte macrophage-colony-stimulating factor (GM-CSF), and monocyte chemoattractant protein (MCP)-1 and also invasion mediators matrix metalloproteinase (MMP)-7, MMP-9, and intracellular adhesion molecule-1. Indeed, TBP inhibits the TNF-alpha-induced expression of all the above endometriotic genes in 12Z endometriotic epithelial cells. The secretion of IL-6, IL-8, GMCSF, and MCP-1 by TNF-alpha is blocked by TBP. Interestingly, MEK, p38, and IKK inhibitors block TNF-alpha-induced IL-8, IL-6, and GM-CSF secretion and 12z invasion, whereas the PI3K inhibitors do not. The only inhibitor to block MCP-1 expression is the p38 inhibitor. Last, TBP, MEK inhibitor, or p38 inhibitor also block cell surface expression of N-cadherin, a marker of mesenchymal cells. Taken together, these results demonstrate that interruption of TNF-alpha-induced signaling pathways in human endometriotic epithelial cells results in decreased expression and secretion of biomarkers for inflammation, EMT, and disease progression.

Topics: Cadherins; Cell Line; Chemokine CCL2; Endometriosis; Epithelial Cells; Extracellular Matrix; Female; Granulocyte-Macrophage Colony-Stimulating Factor; Humans; I-kappa B Kinase; Inflammation; Inflammation Mediators; Interleukin-6; Interleukin-8; MAP Kinase Signaling System; Mesoderm; NF-kappa B; p38 Mitogen-Activated Protein Kinases; Phosphoinositide-3 Kinase Inhibitors; Protein Kinase Inhibitors; Proto-Oncogene Proteins c-akt; Receptors, Tumor Necrosis Factor, Type I; Transcription, Genetic; Tumor Necrosis Factor-alpha

Metformin, a widely used treatment for diabetes that improves insulin sensitivity, also has both antiinflammatory properties and a modulatory effect on ovarian steroid production, two actions that have been suggested to be efficacious in therapy for endometriosis.. To determine whether metformin may be effective for the treatment of endometriosis, we evaluated the effects of this agent on inflammatory response, estradiol production, and proliferation of endometriotic stromal cells (ESCs).. ESCs derived from ovarian endometriomas were cultured with various concentrations of metformin.. IL-8 production, mRNA expression and aromatase activity, and 5-bromo-2'-deoxyuridine incorporation in ESCs were measured.. Metformin dose-dependently suppressed IL-1beta-induced IL-8 production, cAMP-induced mRNA expression and aromatase activity, and 5-bromo-2'-deoxyuridine incorporation in ESCs.. These results suggest that further investigation into the unique therapeutic potential of metformin as an antiendometriotic drug is warranted.

Topics: Adult; Antimetabolites; Aromatase; Biomarkers; Bromodeoxyuridine; Cell Separation; Cell Survival; Cells, Cultured; Endometriosis; Endometrium; Enzyme Activation; Estrogens; Female; Humans; Hypoglycemic Agents; Inflammation; Interleukin-1beta; Interleukin-8; L-Lactate Dehydrogenase; Metformin; Middle Aged; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Stromal Cells

Molecular abnormalities in the epithelial cells of endometriosis and their relevance to carcinogenesis of the ovary have been well studied. On the other hand, the differences of proinflammatory microenvironments between endometriosis and ovarian carcinomas have not been well documented yet. In this study, the expression patterns of CXC chemokines (IL-8, ENA-78, GRO-alpha, I-TAC, Mig, and SDF-1) and their receptors (CXCR2, CXCR3, and CXCR4) were compared among 12 ovarian carcinomas, 8 endometriosis, and 6 normal ovaries using quantitative reverse transcriptase polymerase chain reaction and immunohistochemistry. The CXCR3-mediated signaling in ovarian carcinoma cells in vitro was also investigated. In quantitative reverse transcriptase polymerase chain reaction, ENA-78 was up-regulated both in endometriosis and carcinomas, whereas I-TAC was detected exclusively in carcinomas. CXCR3 was up-regulated both in carcinomas and endometriosis. However, immunohistochemical studies revealed that the localization of CXCR3 in carcinomas was distinctively different from that in endometriosis. In carcinoma-endometriosis coexisting cases, CXCR3-positive lymphocytes in benign lesions decreased in proportion as CXCR3-positive tumor cells replaced the tissues. CXCR3 was also detected in ovarian carcinoma cell lines in vitro. Administration of interferon gamma (IFN-gamma)-inducible chemokines induced extracellular signal-regulated kinase phosphorylation in these carcinoma cells. The results indicated that CXC chemokines might contribute to the progression of ovarian carcinomas and endometriosis in different manners. Aberrant expression of IFN-gamma-inducible chemokines and CXCR3 in carcinoma cells in association with reduced CXCR3-positive immune cells raised the possibility that IFN-gamma-inducible chemokines might not exert effective antitumor immune responses but that they might work in favor of tumor progression.

Topics: Adult; Aged; Cell Line, Tumor; Chemokine CXCL1; Chemokine CXCL11; Chemokine CXCL12; Chemokine CXCL5; Chemokine CXCL9; Chemokines, CXC; Endometriosis; Female; Humans; Immunohistochemistry; Interleukin-8; Middle Aged; Ovarian Neoplasms; Ovary; Receptors, CXCR; Receptors, CXCR3; Receptors, CXCR4; Up-Regulation

Chemokines play an important role in the pathogenesis of endometriosis. In the present study, the transcription of 18 chemokine receptors in eutopic endometrium and ectopic tissue with endometriosis was first analysed by RT-PCR. Dioxin, an air pollutant, and estrogen are reported to be associated with endometriosis. The regulatory mechanisms of dioxin and estrogen in the expression of CXCR1/IL-8 in the corresponding cells will help in elucidating roles of the chemokine in the aetiology of endometriosis.. CXCR1, a type of chemokine receptor, was over-expressed in endometriotic tissue. The high translation of the receptor and its ligand, interleukin (IL-8), in endometriotic tissue was then demonstrated by immunochemistry. Estradiol and 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) alone inhibited expression of CXCR1, whereas the combination of estradiol with TCDD up-regulated the expression. TCDD promoted IL-8 secretion by human pelvic mesothelial cells (HPMC), and 17beta-estradiol magnified the stimulatory effect. Both 17beta-estradiol and TCDD alone inhibited IL-8 secretion of U937 (a cell line of monocyte), but combination of 17beta-estradiol and TCDD had no further inhibitory effect. The co-culture of endometrial stromal cells (ESC) with HPMC produced more IL-8 than respective or total production of either of the cells alone, and estradiol played a synergistic stimulatory role with TCDD in IL-8 secretion of the co-culture. Interaction of HPMC and the monocytes significantly stimulated IL-8 secretion, suggesting a main resource of IL-8 in peritoneal cavity with endometriosis. TCDD promoted IL-8 secretion by HPMC-U937 co-culture, but exerted a contrary effect for IL-8 secretion when combined with estradiol.. Estradiol and TCDD in the peritoneal cavity can lead to a persistent and serious inflammation, which gives a new insight into the interactions of estrogen and TCDD in endometriosis.

Topics: Adult; Cells, Cultured; Coculture Techniques; Drug Synergism; Endometriosis; Endometrium; Environmental Pollutants; Estradiol; Female; Gene Expression Regulation; Humans; Immunohistochemistry; Interleukin-8; Middle Aged; Polychlorinated Dibenzodioxins; Receptors, Interleukin-8A; Signal Transduction; U937 Cells

An increase in the level of the vascular endothelial growth factor (VEGF) production has been reported in the peritoneal fluid (PF) of endometriosis patients. This suggests that changes in the vascular permeability and angiogenesis play an important role in the pathophysiology of this disease. This study examined the effects of the PF obtained from endometriosis patients on the release of VEGF by neutrophils and monocytes.. Neutrophils and monocytes were obtained from young healthy volunteers and cultured with the PF obtained from either endometriosis patients (EPF) (n=18) or a control group (CPF) (n=4). A human monocyte/macrophage cell line, THP-1, was cultured with either 10% EPF or 10% CPF. The PF and culture supernatants were assayed for VEGF using ELISA. Real-time PCR and Western blotting were used to measure the VEGF mRNA and protein expression level, respectively.. The VEGF levels were higher in the EPF than in the CPF (591+/-75 versus 185+/-31 pg/ml, P<0.05). However, the level of VEGF released by THP-1 cells in CPF and EPF was similar. The EPF induced the release of VEGF by neutrophils, but no VEGF was released by monocytes. The VEGF mRNA expression levels in the neutrophils were higher in the EPF, which was abrogated by cycloheximide, suggesting that the EPF induces the production of VEGF in neutrophils. Neutralizing antibodies against IL-8 and TNF-alpha did not completely prevent the EPF-induced release of VEGF by the neutrophils, even though these growth factors stimulated the release of VEGF by neutrophils. There was a positive correlation between the VEGF and IL-10 concentrations in the EPF (correlation coefficient=0.549, P=0.012, n=18), but the neutralizing antibody of IL-10 did not affect the release of VEGF by the EPF-treated neutrophils.. The EPF induced the production and release of VEGF by neutrophils, suggesting that neutrophils may be a source of peritoneal VEGF. In addition, neutrophil-derived VEGF might be a marker for diagnosing endometriosis.

Topics: Ascitic Fluid; Cell Line; Cell Survival; Endometriosis; Female; Humans; Interleukin-10; Interleukin-8; Monocytes; Neutrophils; RNA, Messenger; Tumor Necrosis Factor-alpha; Vascular Endothelial Growth Factor A

The purpose of this study was to evaluate whether pregnancy-associated plasma protein A, glycodelin, osteoprotegerin, and soluble CD163 are possible peritoneal fluid markers for endometriosis and to compare them with the established chemokine markers interleukin-8 and regulated on activation, normal T-cell expressed and secreted.. Determination of the concentrations of interleukin-8, regulated on activation, normal T-cell expressed and secreted, pregnancy-associated plasma protein A, glycodelin, CD163, osteoprotegerin, and progesterone in the peritoneal fluid collected from women undergoing laparoscopy.. From a total of 132 women, 77 women were diagnosed with endometriosis, and 55 women were free of the disease and served as control subjects. Pregnancy-associated plasma protein A and osteoprotegerin showed significantly (P < 0.05) elevated peritoneal fluid concentrations as a function of the severity of the disease, together with interleukin-8 and regulated on activation, normal T-cell expressed and secreted (P < .001). Glycodelin and CD163 did not differ between cases and control subjects. Many of these marker concentrations were intercorrelated strongly.. Pregnancy-associated plasma protein A and osteoprotegerin may play a role in the inflammation process of endometriosis, but interleukin-8 and regulated on activation, normal T-cell expressed and secreted are superior peritoneal fluid markers.

Topics: Adult; Ascitic Fluid; Chemokine CCL5; Endometriosis; Female; Glycoproteins; Humans; Interleukin-8; Osteoprotegerin; Pregnancy-Associated Plasma Protein-A; Receptors, Cytoplasmic and Nuclear; Receptors, Tumor Necrosis Factor

Topics: Biomedical Research; Endometriosis; Female; Genetic Predisposition to Disease; Humans; Interleukin-8; Laparoscopy; Progestins; Tumor Necrosis Factors

The elevation of the proinflammatory chemoattractant cytokine levels in ectopic and eutopic endometrium of endometriosis implies an inflammatory basis for this disease. The relationship between endothelial cells and leukocytes is likely to be important in the regulation of inflammatory mediators of endometriosis. The aim of this study was to describe the temporal and spatial expression of IL-8 in human endometrial endothelial cells (HEEC) in vivo and to compare the in vitro regulation of IL-8 expression by sex steroids in HEEC from women with or without endometriosis. Eutopic endometrial tissues and endometriosis implants were grouped according to menstrual cycle phase and examined by immunohistochemistry for IL-8 expression. Endothelial cells of endometriotic implants expressed higher IL-8 immunoreactivity compared with endothelial cells of eutopic endometrium from women with or without endometriosis (P < 0.02). For in vitro studies, HEEC were isolated from women with or without endometriosis and grown to preconfluence. The purity of cultured HEEC (90-95%) was confirmed by immunocytochemistry using endothelium-specific markers, CD31 and CD146. The effects of estradiol (5 x 10(-8) m), progesterone (10(-7) m), or both on IL-8 mRNA and protein levels were analyzed by RT-PCR and ELISA, respectively. Sex steroids reduced the expression of IL-8 mRNA and protein in HEEC from women without endometriosis. In contrast, both estradiol and progesterone stimulated IL-8 mRNA and protein expression in HEEC from women with endometriosis. We postulate that the stimulation of chemokine expression by sex steroids in HEEC of women with endometriosis may play a role in the inflammatory aspect of this disease.

Topics: Cells, Cultured; Endometriosis; Endometrium; Endothelial Cells; Estradiol; Female; Gene Expression; Humans; Immunohistochemistry; Interleukin-8; Progesterone; RNA, Messenger; Stromal Cells

Endometriosis, a common disease among women of reproductive age, is characterized by the presence of endometrial-like tissue outside the uterus. TNF-alpha induces IL-8 production in endometriotic cells through nuclear factor-kappaB (NF-kappaB) activation. Thalidomide (Thal) inhibits inflammation by down-regulating the expression of proinflammatory cytokines in tumor cells and inflammatory cells. However, the mechanism of Thal action in human endometriotic stromal cells has not yet been elucidated.. We examined whether Thal abrogates TNF-alpha-induced up-regulation of IL-8 expression in endometriotic stromal cells.. Here, we show 1) that treatment of endometriotic stromal cells with TNF-alpha increased the expression of phosphorylated IkappaBalpha and degradation of total IkappaBalpha, which in turn activates NF-kappaB; 2) Thal significantly inhibits the TNF-alpha-induced expression of phosphorylated IkappaBalpha and degradation of IkappaBalpha; 3) TNF-alpha activation induced increased nuclear translocation of NF-kappaB, which was inhibited by pretreatment with either Thal or N-tosyl-L-phenylalanine chloromethyl ketone, an NF-kappaB inhibitor. Thal did not enhance the N-tosyl-L-phenylalanine chloromethyl ketone's action; and 4) Pretreatment with Thal reduced TNF-alpha-induced IL-8 protein production as well as mRNA expression.. The current study showed for the first time that Thal treatment attenuated the expression of IL-8 by reducing TNF-alpha-induced NF-kappaB activation.

Topics: Active Transport, Cell Nucleus; Cells, Cultured; Endometriosis; Female; Humans; I-kappa B Proteins; Interleukin-8; NF-KappaB Inhibitor alpha; Stromal Cells; Thalidomide; Tosylphenylalanyl Chloromethyl Ketone; Tumor Necrosis Factor-alpha

Endometriosis is known to be associated with local inflammatory reactions. Given the emerging concept of thrombin and its specific receptor, protease-activated receptor 1 (PAR1), as important players in inflammation and cell proliferation, we investigated whether thrombin and PAR1 might be involved in the pathophysiology of the disease, using a primary cell culture system of endometriotic tissues. PAR1 mRNA was expressed in primary endometriotic stromal cells (ESCs). Thrombin and SFLLRN (Ser-Phe-Leu-Leu-Arg-Asp), a PAR1 agonist peptide, increased the mRNA expression of IL-8, monocyte chemoattractant protein-1 (MCP-1), and cyclooxygenase-2 (COX-2) and the protein secretion of IL-8 nd MCP-1 in ESCs. The addition of thrombin inhibitor d-phenylalanyl-l-prolyl-l arginine chloromethyl ketone (PPACK) together with thrombin inhibited the thrombin-induced secretion of IL-8 and MCP-1. Thrombin, but not SFLLRN, activated matrix metalloproteinase-2 in ESCs, and the effect was inhibited by PPACK. Thrombin and SFLLRN increased proliferating cell nuclear antigen-positive ratio of ESCs, indicating their cell proliferation-stimulating effects. The thrombin-induced increase in proliferating cell nuclear antigen-positive ratio was diminished by PPACK. These findings imply that the thrombin system might be involved in the pathophysiology of endometriosis, stimulating inflammatory responses of endometriotic cells and their mitogenic activity.

Topics: Chemokine CCL2; Cyclooxygenase 2; DNA Primers; Endometriosis; Female; Humans; Interleukin-8; Membrane Proteins; Prostaglandin-Endoperoxide Synthases; Receptor, PAR-1; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Thrombin

To clarify the inflammatory nature of adenomyosis, we aimed to investigate the expression of interleukin-8 (IL-8) and monocyte chemotactic protein-1 (MCP-1) by immunohistochemistry to determine their putative role in pathophysiology of adenomyosis.. Adenomyosis samples, with their eutopic endometrium, were collected from 30 women undergoing hysterectomy. Endometrium from 27 women without adenomyosis were also collected as a control group. Samples were grouped according to the menstrual cycle phase and examined by immunohistochemistry for IL-8 and MCP-1.. In normal endometrium, secretory phase samples expressed higher levels of epithelial IL-8 than in proliferative phase samples (P = 0.01), and we observed a trend for an increased epithelial MCP-1 expression in the secretory phase samples compared with the proliferative phase samples (P = 0.07). Endometrial samples of women with adenomyosis did not show the same cyclic variation. In the secretory phase, eutopic endometrium of women with adenomyosis expressed lower levels of epithelial IL-8 and MCP-1 compared with normal endometrium (P < 0.05). The expression of epithelial IL-8 and MCP-1 was higher in the adenomyosis foci than the eutopic endometrium (P < 0.05).. These findings may indicate that an intrinsic abnormality of inflammatory response may be present in eutopic endometrium of women with adenomyosis, and IL-8 and MCP-1 may contribute to the pathophysiology of adenomyosis.

Topics: Adult; Chemokine CCL2; Endometriosis; Endometrium; Female; Gene Expression Regulation; Humans; Hysterectomy; Immunohistochemistry; Inflammation; Interleukin-8; Middle Aged; Retrospective Studies; Time Factors

GnRH II is the second form of GnRH and is widely distributed in peripheral tissues of the female reproductive tract as well as in the central nervous system. In the present study, we studied the possible implication of GnRH II in endometriosis.. Effects of GnRH II on 5-bromo-2'-deoxyuridine (BrdU) uptake by cultured endometriotic stromal cells were examined. Effects of GnRH II on interleukin (IL)-1beta-induced expression of cyclooxygenase (COX)-2 and IL-8 were also studied. mRNA levels of GnRH I, GnRH II, type I GnRH receptor and type II GnRH receptor were determined by real-time quantitative RT-PCR in endometrial tissues of women with or without endometriosis and in endometriotic tissues.. GnRH II dose-dependently suppressed BrdU uptake by endometrial stromal cells. Treatment with IL-1beta markedly increased mRNA levels of COX-2 and IL-8 in endometrial stromal cells and IL-8 protein secretion by these cells, while these increments were significantly suppressed by supplementation with GnRH II. The mRNA levels of GnRH II were lower in endometrial and endometriotic tissues of women with endometriosis than in endometrial tissues of women without endometriosis, both in the proliferative phase and the secretory phase. In addition, as for GnRH I, type I GnRH receptor and type II GnRH receptor, the mRNA levels were lower in endometrial tissues of women with endometriosis than in those without endometriosis in the secretory phase.. In the light of the demonstrated antiproliferative and anti-inflammatory effects of GnRH II on endometrial stromal cells, the lower expression of GnRH II in eutopic and ectopic endometrium of women with endometriosis suggests that endogenous GnRH II-mediated cytostatic regulation may be impaired in the development of endometriosis.

Topics: Adult; Bromodeoxyuridine; Cell Proliferation; Cells, Cultured; Cyclooxygenase 2; Endometriosis; Endometrium; Female; Gonadotropin-Releasing Hormone; Humans; Interleukin-1; Interleukin-8; Receptors, LHRH; RNA, Messenger; Stromal Cells

Inflammation has been proposed to play essential roles in the pathophysiology of endometriosis, in which neutrophils and mast cells have been suggested to be involved. We studied whether the protease-activated receptor 2 (PAR2), which is activated by enzymes from neutrophils and mast cells, in endometriotic stromal cells (ESC) has any implication in the development of the disease.. Cultured ESC were stimulated with various concentrations of a specific PAR2 agonist peptide. Proliferating activity of the cells was determined using immunostaining of proliferating cell nuclear antigen (a cell proliferation marker), 5-bromo-2'-deoxyuridine incorporation into DNA and cell count. The concentrations of interleukin (IL)-6 and IL-8 were measured using specific enzyme-linked immunosorbent assay kits. The phosphorylation of three mitogen-activated protein kinases (MAPK), i.e. p38 MAPK, p42/44 MAPK and stress-activated protein Kinase/c-jun N terminal Kinase, in ESC was examined with Western blot analysis.. Activation of PAR2 stimulated the proliferation of ESC and the secretion of IL-6 and IL-8 from ESC in a dose-dependent manner. Activation of PAR2 stimulated the phosphorylation of all three MAPK, and inhibitors of each MAPK suppressed the PAR2 activation-induced proliferation of ESC.. The activation of PAR2 in ESC may be involved in the pathophysiology of endometriosis by inducing the growth and inflammation of endometriotic lesions.

Topics: Blotting, Western; Bromodeoxyuridine; Cell Count; Cell Proliferation; DNA; Dose-Response Relationship, Drug; Endometriosis; Endometrium; Enzyme Activation; Enzyme Inhibitors; Enzyme-Linked Immunosorbent Assay; Female; Humans; Immunohistochemistry; Interleukin-6; Interleukin-8; MAP Kinase Signaling System; Mast Cells; Neutrophils; Phosphorylation; Proliferating Cell Nuclear Antigen; Receptor, PAR-2; Stromal Cells; Time Factors

The aim of this study was to study the regulatory function of Chinese medicine-YiWeiNing (YWN) on the cytokines of endometriosis (EM) model rats.. After successfully creating EM models of rats, all were separated randomly into five groups: high dose of YWN (5.4 g/100 g), low dose of YWN (1.8 g/100 g), Danazol group (3.6 mg/100 g), a model group, and a false-operation group. They were then reviewed for the variation of the amount of TNF-alpha (tumor necrosis factor-alpha), interleukin-6 (IL-6), and interleukin-8 (IL-8).. The content of TNF-alpha, IL-6, and IL-8 in the peripheral blood of the model group was apparently higher than the false-operation group (p < 0.01), and YWN reduced the amount of TNF-alpha, IL-6, and IL-8 in the serums of the model group's rats. This made these cytokines tend toward normal levels without an apparent variation, when compared with the false-operation group (p > 0.05).. YWN can prevent the growth of ectopic endometrium by inhibiting the synthesis and secretion of TNF-alpha, IL-6, and IL-8.

Topics: Animals; Cytokines; Drugs, Chinese Herbal; Endometriosis; Enzyme-Linked Immunosorbent Assay; Female; Interleukin-6; Interleukin-8; Models, Animal; Phytotherapy; Random Allocation; Rats; Time Factors; Tumor Necrosis Factor-alpha

To evaluate cytokine levels (IL-1beta, TNF-alpha, IL-6, IL-8), soluble intercellular adhesion molecule-1 (sICAM-1) and number of macrophages in peritoneal fluid (PF) of women with no minimal endometriosis and associated (or not) infertility in order to discriminate between infertility and/or endometriosis.. Cytokines and sICAM-1 were measured by using quantitative enzyme-linked immunosorbent assay (ELISA) while the macrophages were identified by May-Grunwald-Giemsa and non-specific esterase staining and presented as medians. The measurements were performed in 78 women belonging to four selected subgroups according to their endometriosis and/or infertility status. Statistical analysis was performed using Kruskal-Wallis non-parametric ANOVA test. Additionally, discriminant function analyses were performed.. We have found the most elevated macrophage numbers in the groups of women with endometriosis. IL-1beta did not present any statistically significant values differentiating the analysed subgroups. IL-6 (110.0 pg/ml) and TNF-alpha exhibited the highest concentrations (statistically significant) in a group of fertile women with endometriosis. IL-8 clearly differentiated between the subgroups with infertility and sICAM-1 was statistically significantly elevated in the subgroups of infertile women. In the forward discriminant analysis, when subdividing the studied population according to its infertility status (we considered macrophages, IL-8 and IL-6 in order of probability values), we have found striking probability value of 92% for the correct classification into an infertile population.. Out of the range of the analysed factors we have found only the sICAM-1 to be singled out between the standard discriminant analysis and the forward one. However, this important flagging molecule might be of considerable value for discrimination between different types of pathology at the level of immune effector function. The increased levels of TNF-alpha and IL-6 signified a group of fertile women with endometriosis; however only IL-6 presented a discriminating value in multifunctional analysis of examined subgroups. The analysed range of factors had a greater tendency to discriminate between infertility status rather than endometriosis.

Topics: Adult; Ascitic Fluid; Biomarkers; Case-Control Studies; Cytokines; Endometriosis; Enzyme-Linked Immunosorbent Assay; Female; Humans; Infertility, Female; Intercellular Adhesion Molecule-1; Interleukin-1; Interleukin-6; Interleukin-8; Tumor Necrosis Factor-alpha

To study the adjustment of Xianggui pill on the cytokine of endometriosis model rat, and investigate the mechanism of Xianggui pill on the treatment of endometriosis.. To set up endometriosis model by rat self-endometria transplantation, drench sodium chloride, Xianggui pill elixation or Danazol after grouping, and to detect the contents of interleukin-8 (IL-8) and tumor necrosis factor-alpha (TNF-alpha) by ELISA.. The contents of IL-8, TNF-alpha in the peripheral blood and peritoneal fluid of model group were higher than that of the blank group; The quality of allotopia growth intima tissue, the quantity of macrophage in peritoneal fluid and the contents of IL-8, TNF-alpha in the Xianggui pill group and Danazol group were all lower than those of the model group; but there was no significant difference of each target between the Xianggui pill group and Danazol group.. Xianggui pill can restrain significantly the growth of allotopia intima tissue, and has apparently adjustment to the cytokine.

Topics: Animals; Cell Count; Drug Combinations; Drugs, Chinese Herbal; Endometriosis; Female; Interleukin-8; Macrophages, Peritoneal; Plants, Medicinal; Random Allocation; Rats; Rats, Sprague-Dawley; Tumor Necrosis Factor-alpha

To assess the release of epithelial neutrophil-activating peptide-78 (ENA-78) into peritoneal fluid in women with and without endometriosis.. Retrospective study.. Nagoya City University Hospital.. Surgery was scheduled in the proliferative or secretory phase of the menstrual cycle for 59 women with (n = 35) and without (n = 24) endometriosis.. Peritoneal fluid samples were obtained at laparotomy or laparoscopy.. The ENA-78 concentrations in the peritoneal fluid were measured using enzyme-linked immunosorbent assay (ELISA).. The concentrations of ENA-78 in the peritoneal fluid were markedly elevated in the endometriosis patients as compared with the controls, especially in women with severe stage disease.. We conclude that ENA-78 is an important factor that may contribute to the pathogenesis of endometriosis, possibly promoting neovascularization.

Topics: Ascitic Fluid; Biomarkers; Chemokine CXCL5; Chemokines, CXC; Chemotaxis, Leukocyte; Endometriosis; Endometrium; Female; Humans; Interleukin-8; Neovascularization, Pathologic; Reference Values; Retrospective Studies

To evaluate the involvement of chemokines in the pathogenesis of endometriosis, we investigated the expression of CXC chemokines in cultured ovarian endometriotic cyst stromal cells (ECSC), endometrial stromal cells with endometriosis (ESCwE), and normal endometrial stromal cells (NESC). Using ELISA, TNF-alpha significantly enhanced the production of IL-8, growth-related oncogene alpha, and epithelial neutrophil-activating peptide-78 in all cases of ECSC (n = 10), ESCwE (n = 6), and, NESC (n = 10). IL-1beta did not affect the production of these chemokines in eight of 10 cases of ECSC. In contrast, IL-1beta significantly enhanced the expression of these chemokines in all cases of ESCwE (n = 6) and NESC (n = 10). Western blot analysis revealed down-regulation of expression of IL-1 receptor type 1 (IL-1-R1) in all cases of ECSC with low response to IL-1beta (n = 8). In contrast, significant IL-1-R1 expression was detected in all cases of NESC. Although IL-1-R1 expression was detected in all cases of ESCwE (n = 6), its expression in ESCwE tended to decrease compared with that in NESC. Moreover, phosphorylation of inhibitor kappaB-alpha was detected in all cases of ESCwE and NESC after stimulation with IL-1beta, but not in ECSC with low response to IL-1beta (n = 8). In contrast, significant IL-1-R2 expression was detected in all cases of ECSC, ESCwE, and NESC. The present findings suggest that the dysregulation of IL-1/IL-1-R system relates to immunological dysfunction in endometriosis. The alteration of the CXC chemokines expression may be important for elucidation of the pathogenesis of endometriosis.

Topics: Chemokine CXCL1; Chemokine CXCL5; Chemokines, CXC; Down-Regulation; Endometrial Stromal Tumors; Endometriosis; Female; Gene Expression; Humans; Intercellular Signaling Peptides and Proteins; Interleukin-8; Leiomyoma; Receptors, Interleukin-1; Receptors, Interleukin-1 Type I; Receptors, Interleukin-1 Type II; Receptors, Tumor Necrosis Factor, Type I; RNA, Messenger; Signal Transduction; Stromal Cells; Tumor Cells, Cultured

To evaluate the effect of lipopolysaccharide (LPS) on the expression of tumor necrosis factor alpha (TNFalpha) and interleukin-8 (IL-8) protein in endometriotic stromal cells (ESC) and their effect on the proliferation of ESC.. Prospective study.. Department of Obstetrics and Gynecology, Tottori University Hospital, Yonago, Japan.. Seventeen patients who underwent laparoscopic surgery.. Endometriotic stromal cells were obtained from chocolate cyst linings of the ovary.. We determined the effect of LPS on the production of TNFalpha and IL-8 and the effect of IL-8 antisense oligonucleotide and nuclear factor-kappaB (NF-kappaB) inhibitor on IL-8 production using ELISA. TNFalpha production was examined by immunocytochemical staining. We determined the effect of LPS and the effect of IL-8 antisense oligonucleotide and NF-kappaB inhibitor on LPS-promoted ESC proliferation.. LPS-stimulated ESC produced significant amounts of TNFalpha and IL-8 in a dose- and time-dependent fashion. Adding LPS promoted ESC proliferation. Anti-TNFalpha antibody and anti-IL-8 antibody inhibited the stimulatory effects of LPS. IL-8 antisense oligonucleotide and NF-kappaB inhibitor significantly decreased LPS-induced IL-8 protein production and LPS-induced ESC proliferation.. Pelvic inflammation may promote the progression of endometriosis.

Topics: Cell Division; Dose-Response Relationship, Drug; Endometriosis; Female; Gene Expression; Humans; Immunohistochemistry; Interleukin-8; Lipopolysaccharides; Membrane Glycoproteins; NF-kappa B; Oligonucleotides, Antisense; Prospective Studies; Receptors, Cell Surface; Staining and Labeling; Stromal Cells; Toll-Like Receptors; Tosylphenylalanyl Chloromethyl Ketone; Tumor Necrosis Factor-alpha

Endometriosis accompanies local inflammatory reactions in the peritoneal cavity. We examined the phosphorylation of mitogen-activated protein kinases (MAPKs), i.e. extracellular signal-regulated kinase (ERK), p38 MAPK (p38) and c-Jun N-terminal kinase (JNK) in endometriotic stromal cells, and their possible pathophysiological roles in endometriosis in relation to proinflammatory substances.. Endometriotic stromal cells were isolated from endometriomas and were cultured for the experiments. Phosphorylation of MAPKs in endometriotic stromal cells treated with interleukin (IL)-1beta, tumor necrosis factor (TNF)alpha and H(2)O(2) were examined by Western blot analysis. Effects of PD98059, SB202190 and SP600125 (inhibitors of ERK, p38 and JNK, respectively) on IL-1beta-induced secretion of IL-6 and IL-8, and on IL-1beta-induced expression of cyclo-oxygenase-2 (COX-2) in endometriotic cells were studied. In addition, eutopic endometrial tissues were collected, and the phosphorylation rate of p38 in eutopic endometrial tissues and endometriotic tissues were determined.. IL-1beta, TNFalpha and H(2)O(2) stimulated the phosphorylation of ERK, p38 and JNK, while the total amounts of proteins of the respective MAPKs were virtually the same compared with those in the unstimulated controls. Both SB202190 and SP600125 suppressed IL-1beta-induced secretion of IL-6 and IL-8, and PD98059 suppressed IL-1beta-induced secretion of IL-8. Both SB202190 and PD98059 suppressed IL-1beta-induced expression of COX-2 in endometriotic cells. The p38 phosphorylation rates in the endometriotic tissues were significantly higher than those in the eutopic endometrial tissues of the same patients.. Given the current theory that inflammatory changes are involved in the progression of endometriosis, MAPKs could play as pivotal intracellular signal transducers in endometriotic cells, and thus have a pathophysiological role in the disease.

Topics: Anthracenes; Cells, Cultured; Cyclooxygenase 2; Endometriosis; Endometrium; Enzyme Inhibitors; Extracellular Signal-Regulated MAP Kinases; Female; Flavonoids; Gene Expression Regulation, Enzymologic; Humans; Hydrogen Peroxide; Interleukin-1; Interleukin-6; Interleukin-8; JNK Mitogen-Activated Protein Kinases; Membrane Proteins; p38 Mitogen-Activated Protein Kinases; Phosphorylation; Prostaglandin-Endoperoxide Synthases; RNA, Messenger; Signal Transduction; Stromal Cells; Tumor Necrosis Factor-alpha

Endometriosis, a common disease among women of reproductive age, is characterized by the presence of endometrial-like tissue outside the uterus. We previously reported that TNFalpha promoted proliferation of endometriotic stromal cells by inducing IL-8 gene and protein expression. We hypothesize that TNFalpha may induce IL-8 production in endometriotic cells through nuclear factor-kappa B (NF-kappa B) activation. Western blot analyses and electrophoretic mobility shift assays revealed that incubation with TNF alpha induced the expression of phosphorylated inhibitor kappa B (p-I kappa B) and activation of NF-kappa B in endometriotic stromal cells. The NF-kappa B inhibitor, N-tosyl-L-phenylalanine chloromethyl ketone, reduced TNFalpha-induced IL-8 gene and protein expression. The medical treatment of endometriosis with GnRH agonist (GnRHa) has been shown to induce hypoestrogenemia and reduce the observable number of endometriotic implants. We compare the expression of IL-8 gene and protein in endometriotic stromal cells of patients treated with GnRHa and those of patients without treatment before laparoscopic cystectomy for endometrioma. The addition of TNFalpha (0.1 ng/ml) significantly increased protein and gene expression of IL-8 in the cells of patients without GnRHa treatment, but this expression was not observed in the cells of patients with GnRHa. The addition of estradiol (E2; 10(-7) M) enhanced the expression of IL-8. However, in the cells of patients who received GnRHa treatment, TNFalpha and E2 did not show any significant effect. In endometriotic stromal cells without GnRHa treatment, TNFalpha and E2 increased the expression of p-I kappa B. In contrast, TNFalpha and E2 had no significant effect on the expression of p-I kappa B in cells that received GnRHa treatment. These findings demonstrate that NF-kappa B activation is critical for TNFalpha-induced IL-8 expression in endometriotic stromal cells. The current study showed for the first time that GnRHa treatment attenuated the expression of IL-8 by reducing TNFalpha-induced NF-kappa B activation.

Topics: Adult; Antineoplastic Agents; Cells, Cultured; Electrophoretic Mobility Shift Assay; Endometriosis; Estrogens; Female; Gene Expression; Gonadotropin-Releasing Hormone; Humans; I-kappa B Proteins; Interleukin-8; NF-kappa B; NF-KappaB Inhibitor alpha; Phosphorylation; Protein Synthesis Inhibitors; RNA, Messenger; Stromal Cells; Tosylphenylalanyl Chloromethyl Ketone; Tumor Necrosis Factor-alpha

Previous evaluations of the relationship between the concentrations of interleukin-8 (IL-8) in the peritoneal fluid and endometriosis led to non-consistent results. Our purpose was to investigate the correlation of the concentrations of IL-8 in the peritoneal fluid with the stage of endometriosis, the presence of red lesions and the phase of the menstrual cycle.. Ninety-two patients with infertility (n = 87) or undergoing sterilization (n = 5) had peritoneal fluid samples collected at laparoscopy. IL-8 determinations were performed using an enzyme-linked immunosorbent assay.. The concentrations of IL-8 in the peritoneal fluid of the 68 women with endometriosis were not significantly different from those of the 24 controls. Patients with moderate/severe stages had IL-8 significantly higher than controls (P = 0.008) and marginally higher than patients with minimal/mild endometriosis (P = 0.053). Concentrations of IL-8 were significantly higher in patients than in controls in the luteal phase. Red lesions were associated with significantly increased levels of peritoneal fluid IL-8 only in the luteal phase.. Our findings reinforce the importance of IL-8 in the pathogenesis of endometriosis.

Topics: Adult; Ascitic Fluid; Case-Control Studies; Endometriosis; Enzyme-Linked Immunosorbent Assay; Female; Humans; Infertility, Female; Interleukin-8; Luteal Phase; Osmolar Concentration; Severity of Illness Index

To determine the contribution of endometrial cells in the development of endometriosis. Specifically the response of the mesothelium to endometrial cells in the production of monocyte chemotactic protein-1 (MCP-1), interleukin-6 (IL-6), and IL-8 was studied.. In vitro study.. University Research Laboratory.. None.. Cellular MCP-1, IL-6 secretion and MCP-1, and IL-6 and IL-8 messenger RNA expression were evaluated by ELISA and reverse transcription-polymerase chain reaction (RT-PCR) assay.. The mesothelial cells produced more MCP-1 and IL-6 than endometrial epithelial and stromal cells. Mesothelial cells cultured in the presence of endometrial epithelial cells produced even greater levels of MCP-1 and IL-6 than those cultured in the presence of stromal cells or cultured alone. The MCP-1, IL-6, and IL-8 mRNA expression also increased when mesothelial cells were co-cultured with endometrial epithelial cells.. The results suggest that endometrial epithelial cells may be important in evoking the inflammatory reaction in the peritoneal cavity during retrograde menstruation and that mesothelial cells may play an important role in the chemotaxis of monocytes and in the inflammatory process during the development of endometriosis.

Topics: Chemokine CCL2; Coculture Techniques; Endometriosis; Enzyme-Linked Immunosorbent Assay; Epithelial Cells; Female; Humans; Interleukin-6; Interleukin-8; Peritoneum; Reverse Transcriptase Polymerase Chain Reaction; RNA

To investigate the presence of epithelial neutrophil-activating peptide 78 (ENA-78) in peritoneal fluid of women with and without endometriosis and to identify the cells that produce this inflammatory protein.. Case-control study.. University hospital.. Eighteen women with and 9 women without endometriosis.. ENA-78 protein and mRNA levels were compared among women with and without endometriosis in samples of peritoneal fluid, samples of endometriotic lesions obtained by biopsy during laparoscopy, and peritoneal macrophages. Enzyme-linked immunosorbent assay, reverse transcription polymerase chain reaction, and in situ hybridization methods were used. Secretion of ENA-78 protein by interleukin-1beta-stimulated endometriotic stromal cells and in the media of lipopolysaccharide-stimulated peritoneal macrophages were compared to that in unstimulated cell cultures.. Peritoneal fluid concentrations of ENA-78 were significantly higher in affected women than in controls. Ectopic epithelial and stromal cells and peritoneal macrophages express ENA-78 messenger RNA. Interleukin-1beta stimulation of stromal cell cultures resulted in a 23-fold increase in ENA-78 concentration, and lipopolysaccharide stimulation of peritoneal macrophages increased concentrations by 8-fold.. Levels of ENA-78 are elevated in the peritoneal fluid of women with endometriosis. Ectopic glandular cells, ectopic stromal cells, and peritoneal macrophages express this inflammatory chemokine. Epithelial neutrophil-activating peptide 78 may play an important role in the pathogenesis of endometriosis.

Topics: Ascitic Fluid; Case-Control Studies; Chemokine CXCL5; Chemokines, CXC; Endometriosis; Female; Humans; In Situ Hybridization; Interleukin-1; Interleukin-8; Lipopolysaccharides; Macrophages, Peritoneal; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger

Altered expression of cytokines has been suggested as a specific event for the maintenance and progression of endometriomas. Few data exist on cytokine expression in endometriomas compared with benign and malignant ovarian tumours. Hence, serum and cyst fluid levels of interleukin (IL)-6, IL-8 and tumour necrosis factor-alpha (TNF-alpha) were evaluated in women with endometriomas and compared with those in women with benign or malignant ovarian tumours.. Investigations included immunoradiometric determination of serum and cyst fluid concentrations of IL-6, IL-8 and TNF-alpha in 34 women with endometriomas, 30 women with benign and 13 women with malignant cystic ovarian tumours.. Serum IL-6 levels were higher in ovarian cancer than in endometriomas (P<0.01) or benign tumours (P<0.01). Serum TNF-alpha levels differed between benign tumours and endometriomas (P<0.01), but not between endometriomas and malignant tumours. Cyst fluid levels of IL-8 were higher in endometriomas than in benign tumours (P<0.001) and lower than in malignant tumours (P=0.03). Cyst fluid levels of TNF-alpha differed between malignant tumours and endometriomas (P<0.01) and benign tumours (P<0.01), but not between endometriomas and benign tumours. In the endometriomas group, a positive correlation was found between serum and cyst fluid levels of IL-6 (P=0.003, rho=0.633), and between serum levels of IL-6 and IL-8 (P=0.03, rho=0.415).. Endometriomas were associated with serum TNF-alpha levels similar to those found in women with ovarian cancer, while serum IL-6 levels and cyst fluid IL-8 levels were intermediate between those observed in benign and malignant ovarian tumours.

Topics: Adolescent; Adult; Aged; Aged, 80 and over; Cyst Fluid; Endometriosis; Female; Humans; Interleukin-6; Interleukin-8; Middle Aged; Ovarian Cysts; Ovarian Neoplasms; Tumor Necrosis Factor-alpha

Numerous cytokines and growth factors are synthesized in the endometrium. IL-8 is one of these cytokines regulating endometrial function. It is a neutrophil chemoattractant/ activating factor and a potent angiogenic agent. IL-8 is elevated in the peritoneal fluid of women with endometriosis. We have previously demonstrated a direct proliferative effect of IL-8 on endometrial stromal cells. We hypothesized that increased levels of IL-8 in the endometriotic environment could up- regulate Fas ligand (FasL) expression in endometrial cells and may be relevant for the development of a relative local immunotolerance in endometriosis by inducing apoptosis of cytotoxic T lymphocytes. To test our hypothesis, we studied the in vitro regulation of FasL expression and apoptosis by IL-8 in endometrial cells. Western blot analysis in endometrial stromal, glandular, and Ishikawa cells revealed that IL-8 up- regulated FasL protein expression in these cells. By semiquantitative RT-PCR analysis, IL-8 does not alter the expression of either Fas or FasL mRNA levels in these cells. Immunocytochemistry results from endometrial stromal cells treated with IL-8 demonstrated an up-regulation of FasL protein expression. IL-8 decreased apoptosis rate in endometrial stromal cells as evaluated by terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick end-labeling assay. We observed an increased apoptotic rate in Jurkat (T lymphocyte line) cells plated on endometrial stromal cells previously treated with IL-8. We speculate that increased FasL expression by IL-8 may induce apoptosis of T lymphocytes and thus produce a local immunotolerant environment for the development of ectopic implants.

Topics: Adult; Apoptosis; Endometriosis; Endometrium; Fas Ligand Protein; fas Receptor; Female; Gene Expression; Humans; Interleukin-8; Jurkat Cells; Membrane Glycoproteins; Middle Aged; RNA, Messenger; Stromal Cells; T-Lymphocytes

During menstruation endometrial fragments are transported into the peritoneal cavity where they form endometriotic lesions. Angiogenesis is proposed as one of the mechanisms in endometriosis pathogenesis. The aim of the study was to determine the angiogenic activity and interleukin 8 concentrations in peritoneal fluid and sera in endometriosis.. Angiogenesis was determined in cutaneous assay in Balb/c mice; IL-8 concentrations were measured by ELISA test in sera and peritoneal fluid of 32 control and 56 endometriosis patients. Wilcoxon and Mann-Whitney tests and Spearman rank correlations were used in statistical analysis.. Peritoneal fluid and sera from the examined group had higher angiogenic activity and interleukin 8 concentrations. There was correlation found between AFS and neovascularization induced by sera and PF of patients with peritoneal lesions.. Angiogenesis plays an important role in pathogenesis of endometriosis. Although IL-8 takes part in neovascularization, there are other factors modulating angiogenesis in endometriosis.

Topics: Animals; Ascitic Fluid; Endometriosis; Female; Humans; Interleukin-8; Mice; Mice, Inbred BALB C; Neovascularization, Physiologic

Endometriosis is a disorder characterised by presence and growth of endometrial tissue outside the uterus, primarily into the peritoneum. The peritoneal fluid (PF) of women with endometriosis undergoes a number of biological changes, including local inflammatory-reparative phenomena and peripheral blood mononuclear cells (PBMC) involvement. These activated cells as well as the endometriotic cells secrete various cytokines with pleiotropic biological activities. Dynamic interplay among cytokines may contribute to realise a favourable microenvironment for the implantation of endometrial cells and the progression of the disease. In the present study, we evaluated the levels of cytokines, such as the tumour necrosis factor-alpha (TNF-alpha), transforming growth factor-beta (TGF-beta), interleukin-8 (IL-8) and monocyte chemotactic protein-1 (MCP-1) in PF and in serum (S) of women with endometriosis to compare their behaviour in both biological fluids. The patients (n = 26) were women of reproductive age attending our observation centre for infertility, diagnosed endometriosis at laparoscopy. Control group (n = 5) consisted of women affected by non-immunologic infertility, diagnosed by explorative laparoscopy. S samples were obtained from peripheral blood before anaesthesia and laparoscopy. PF samples were collected at the time of laparoscopy. Both biological fluids were examined for cytokine by ELISA assays. Our results showed that S and PF levels of TNF-alpha, not dosable in controls, were very high at the early stage and decreased significantly with the severity of the disease (p < 0.001). TGF-beta levels were significantly (p < 0.001) higher than in controls and increased with the severity of the disease (p < 0.001), particularly in the PF. S and PF IL-8 as well as MCP-1 concentrations at all stages were higher than in controls (p < 0.001), yet showed an opposite behaviour in both biological fluids. In fact, S levels of IL-8 and MCP-1 were significantly (p < 0.001) higher at early stages and decreased with the severity of the disease, whereas we observed a significant (p < 0.001) enhancement of these chemokine levels in PF from stage I to stage II and stage III. These observations showed that TNF-alpha and TGF-beta levels were overlapping in S and PF of women with endometriosis. On the contrary, MCP-1 and IL-8 S concentrations decreased with the severity of the disease, whereas PF levels showed markedly increased at severe stages. Taken together the observe

Topics: Adult; Ascitic Fluid; Case-Control Studies; Chemokine CCL2; Cytokines; Endometriosis; Enzyme-Linked Immunosorbent Assay; Female; Humans; Interleukin-8; Severity of Illness Index; Transforming Growth Factor beta; Tumor Necrosis Factor-alpha

To study the affects of interleukin-8 (IL-8), anti-IL-8, and IL-12 on in vitro proliferation of endometrial cells.. An in vitro study.. Department of Obstetrics and Gynecology, University of Aberdeen, UK.. Women attending a fertility clinic.. In vitro cell cultures using culture media supplemented with IL-8 (100 ng/mL, 200 ng/mL, and 500 ng/mL), IL-12 (1 ng/mL, 5 ng/mL, and 25 ng/mL), and anti-IL-8 (0.1 microg/mL, 1 microg/mL, 10 microg/mL).. In vitro survival of dispersed endometrial cells (combined epithelial and glandular) at 5 and 9 days of culture.. There was a dose-dependent stimulatory effect of IL-8 on survival of cells. From women with and without endometriosis, IL-12 at 1 ng/mL significantly inhibited the survival of endometrial cells from women without endometriosis as compared with cells from women with endometriosis. At 1 microg/mL, anti-IL-8 significantly inhibited the survival of endometrial cells from women with endometriosis compared with cells from women without endometriosis on day 5 of culture.. Our findings confirm the stimulatory effects of IL-8 and its possible role in the pathogenesis of endometriosis. The effects of IL-12 and anti-IL-8 on endometrial cell survival varied according to the disease state and the concentration of the cytokines. Future in vitro studies on the role of anti-IL-8 and IL-12 should aim to use a greater range of concentrations and a higher density of endometrial cells in cultures supplemented with monocytes.

Topics: Cell Culture Techniques; Cells, Cultured; Dose-Response Relationship, Drug; Endometriosis; Endometrium; Female; Humans; Interleukin-12; Interleukin-8; Luteal Phase; Stromal Cells

The increased production of pro-inflammatory chemoattractant cytokines for neutrophils in endometriosis suggests that changes in the immune system play an important role in the pathophysiology of endometriosis. The effects of plasma and peritoneal fluid from patients with advanced endometriosis on the apoptosis of neutrophils were investigated.. Apoptotic changes of neutrophils were evaluated by morphological changes using Giemsa staining. Apoptosis was confirmed by DNA electrophoretic analysis.. Compared with the plasma (n = 20) and peritoneal fluid (n = 5) of healthy controls, the addition of 10% plasma (n = 20) and peritoneal fluid (n = 10) from patients with endometriosis to an in-vitro culture of neutrophils from healthy subjects reduced the percentage of apoptotic cells from 65.3 +/- 6.6 to 27.2 +/- 4.6% (P < 0.001) and from 45.3 +/- 4.8 to 10.5 +/- 4.3% (P < 0.001) respectively. Neutralizing interleukin-8 antibody abrogated the delay of neutrophil apoptosis induced by peritoneal fluid, but not in the plasma of endometriosis patients.. These findings show that interleukin-8 is one of the neutrophil survival factors in the peritoneal fluid of endometriosis patients and that an unidentified survival factor is also present in the plasma of patients with endometriosis.

Topics: Adult; Antibodies; Apoptosis; Ascitic Fluid; Blood Physiological Phenomena; Cells, Cultured; Dactinomycin; Endometriosis; Female; Humans; Interleukin-8; Neutrophils; Protein Synthesis Inhibitors; Reference Values; Time Factors

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.. 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.. 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.. The results obtained indicate a significant role of local cytokine production impairment in the development of adenomyosis.

Topics: Case-Control Studies; Cytokines; Endometriosis; Endometrium; Epidermal Growth Factor; Female; Humans; In Vitro Techniques; Interferon-alpha; Interferon-gamma; Interleukin-1; Interleukin-8; Leukocytes, Mononuclear; Tumor Necrosis Factor-alpha

To determine the levels of the angiogenic factors vascular endothelial growth factor (VEGF) and interleukin (IL-8) in ovarian cysts.. Prospective descriptive study.. University hospital.. One hundred women, of whom 9 had ovarian carcinomas, 38 had ovarian endometriomata, 43 had serous ovarian cysts, and 10 had follicular ovarian cysts.. Sampling of serum and ovarian cystic fluid before and during surgery.. Levels of VEGF and IL-8 in cystic fluid and serum.. Levels of both VEGF and IL-8 were found to be significantly higher in the cystic fluid of ovarian carcinomas and endometriomata than in serous and follicular cysts. In endometriomata fluid, levels of VEGF and IL-8 were found to be directly correlated (r = 0.68; P=.0074). Serum levels of VEGF were significantly higher in women with ovarian carcinomas and endometriomata than in those with serous and follicular cysts. Ovarian cancers and endometriomata were similar in terms of cystic concentrations of VEGF and IL-8 and in serum levels of VEGF.. An increase in angiogenic factors that differentiate ovarian carcinomas and endometriomata from other kinds of ovarian pathology is demonstrated.

Topics: Carcinoma; Endometriosis; Endothelial Growth Factors; Female; Follicular Cyst; Humans; Interleukin-8; Lymphokines; Osmolar Concentration; Ovarian Cysts; Ovarian Diseases; Ovarian Neoplasms; Prospective Studies; Vascular Endothelial Growth Factor A; Vascular Endothelial Growth Factors

Endometriosis, an oestrogen-dependent disorder affecting women of reproductive age, is associated with active angiogenesis and an increased recruitment of leukocyte into the peritoneal cavity where the implants often develop. The role of oestrogens in the development of endometriosis has been clearly established, but the biochemical mechanisms of their action are still not clearly elucidated. The present study shows that interleukin-1 (IL-1) induces interleukin-8 (IL-8) secretion by endometriotic cells and that oestradiol enhances endometriotic cell responsiveness to IL-1. In contrast, no significant cell responsiveness to progesterone either alone in the culture medium or in combination with oestradiol was noted. Positive immunostaining for IL-8 was observed throughout endometriotic tissue, and no perceptible difference in the intensity of staining regarding the menstrual cycle phase was observed. Together with the in-vitro data, this suggests that IL-8 expression in endometriotic tissue is not subject to cyclic variation. Furthermore, this study provides evidence that oestradiol indirectly up-regulates the expression by ectopic endometrial cells of IL-8, a cytokine endowed with neutrophil chemotactic and angiogenic properties. This may contribute to peritoneal leukocyte recruitment and to the growth of endometriotic implants, and may be a new mechanism for oestradiol action in endometriosis.

Topics: Adolescent; Adult; Cells, Cultured; Endometriosis; Estradiol; Female; Humans; Interleukin-1; Interleukin-8; Menstrual Cycle; Middle Aged; Up-Regulation

To study the effects of omega-3 and omega-6 polyunsaturated fatty acid (PUFA) on in vitro proliferation of endometrial cells and their production of the cytokine interleukin-8 (IL-8).. In vitro study.. Obstetrics and gynecology department, University of Aberdeen.. Women attending an infertility clinic.. In vitro cell cultures using culture mediums supplemented with normal and high ratios of omega-3 PUFA and omega-6 PUFA.. In vitro survival and production of IL-8 by dispersed endometrial cells.. In vitro survival of endometrial cells from women with and without endometriosis was significantly reduced in the presence of high omega-3:omega-6 PUFA ratios compared with cells incubated in the absence of fatty acids, in balanced omega-3:omega-6 PUFA ratios, and in high omega-6:omega-3 PUFA ratios. Endometrial cells from women with endometriosis secreted higher concentrations of IL-8, especially in the presence of high omega-3:omega-6 PUFA ratios.. omega-3 PUFA may have a suppressive effect on the in vitro survival of endometrial cells and omega-3 PUFA be useful in the management of endometriosis by reducing the inflammatory response and modulating cytokine function.

Topics: Cell Survival; Cells, Cultured; Culture Media; Endometriosis; Endometrium; Fatty Acids, Omega-3; Fatty Acids, Omega-6; Fatty Acids, Unsaturated; Female; Humans; Interleukin-8; Reference Values; Stromal Cells

Patients with endometriosis are characterized by the ability of the endometrium to implant and by the peritoneal response to the tissue; angiogenic factors may play a significant role in the aetiology of endometriosis supporting the implantation of ectopic endometrial cells. Vascular endothelial growth factor (VEGF) is a mitogen, morphogen and chemoactractant for endothelial cells and, in vivo, it is a powerful mediator for vessel permeability. Interleukin-8 (IL-8) is a chemoatractant for neutrophils and is a potent angiogenic factor. Women (n = 20) with ovarian endometriomata and 10 women with follicular cysts were enrolled in this study to investigate the role of VEGF and IL-8 in the development and maintenance of ovarian endometriomata. Cystic fluids were collected by laparoscopy, immediately centrifuged and stored until the enzyme-linked immunosorbent assays were performed. The VEGF and IL-8 concentrations were found to be significantly higher in the fluids of the ovarian endometriomata than in those of the follicular cysts of controls (P < 0.001 and P < 0.001 respectively); in addition, a significant inverse correlation between the VEGF cystic fluid concentrations and the diameter of the endometriotic adnexal masses was found (r = -0.56, P = 0.01). The evidence that the high concentrations of VEGF and IL-8 are present in the ovarian endometriomata indicates that angiogenesis could be a specific event both for the progression and maintenance of such cysts.

Topics: Adult; CA-125 Antigen; Endometriosis; Endometrium; Endothelial Growth Factors; Female; Follicular Cyst; Humans; Interleukin-8; Lymphokines; Retrospective Studies; Vascular Endothelial Growth Factor A; Vascular Endothelial Growth Factors

Endometriosis, a common disease among women of reproductive age, is characterized by the presence of endometrial-like tissue outside the uterus. We and others showed that several cytokine levels, including interleukin-8 (IL-8) and tumor necrosis factor-alpha (TNFalpha), are elevated in the peritoneal fluid of women with endometriosis compared with those in women without endometriosis. We also demonstrated that the addition of IL-8 to the culture medium stimulated the proliferation of cultured endometriotic stromal cells. TNFalpha is a multipotent cytokine that induces IL-8 production in various cell types. Therefore, we hypothesized that TNFalpha may also contribute to the pathogenesis of endometriosis by inducing the production of IL-8. To test this hypothesis, we analyzed the peritoneal fluid concentrations of IL-8 and TNFalpha using enzyme-linked immunosorbent assay (ELISA). We observed a significant correlation between the levels of TNFalpha and IL-8 in the peritoneal fluid of endometriosis patients. We also obtained the endometriotic stromal cells from chocolate cyst linings of the ovary. The expression of the receptors for TNFalpha (TNFR) was examined by RT-PCR. We observed the expression of both TNFR-I and TNFR-II genes in endometriotic stromal cells. The expression of IL-8 gene and protein was analyzed by Northern blot hybridization and enzyme-linked immunosorbent assay, respectively. TNFalpha induced the gene and protein expression of IL-8 in endometriotic stromal cells in a dose-dependent fashion. The addition of TNFalpha promoted the proliferation of the endometriotic stromal cells, and the stimulatory effects of TNFalpha were abolished by adding anti-IL-8 antibody. We demonstrated for the first time that TNFalpha stimulated proliferation of endometriotic stromal cells through induction of IL-8 gene and protein expression. We concluded that the TNFalpha may be one of the essential factors for the pathogenesis of endometriosis.

Topics: Adult; Ascitic Fluid; Cell Division; Cells, Cultured; Endometriosis; Female; Gene Expression Regulation; Humans; Interleukin-8; Osmolar Concentration; Receptors, Tumor Necrosis Factor; Stromal Cells; Tumor Necrosis Factor-alpha

Fertilization and oocyte cleavage rates have previously been demonstrated to be lower for women with endometriosis undergoing IVF compared with controls. This might be related to impaired oocyte function, possibly due to an inflammatory milieu in the pelvis of these women, where an elevated concentration of many cytokines is documented. The aim of this study was to examine whether granulosa cells from women with endometriosis deviated with respect to production of the inflammatory cytokines interleukin-1beta, interleukin-6, interleukin-8 and tumour necrosis factor alpha (TNFalpha) compared with granulosa cells from healthy women, undergoing IVF for male infertility. The effect of human chorionic gonadotrophin on cytokine production was also investigated. Granulosa cells in follicular fluid were obtained at oocyte retrieval for IVF. Incubated cell culture media were analysed by enzyme-linked immunosorbent assay. The basal production of all four cytokines was higher in cells from women with endometriosis when compared to controls, although the increase was only significant for TNFalpha. Chorionic gonadotrophin had no significant effect, although it had a tendency to suppress cytokine release in both patient categories. Whether aberrant cytokine production in granulosa cells from women with endometriosis may disturb fertilizing capacity of oocytes requires study.

Topics: Adult; Cells, Cultured; Chorionic Gonadotropin; Cytokines; Endometriosis; Female; Fertilization in Vitro; Granulosa Cells; Humans; Infertility, Female; Infertility, Male; Interleukin-1; Interleukin-6; Interleukin-8; Male; Pregnancy; Pregnancy Rate; Reference Values; Tumor Necrosis Factor-alpha

The production of IL-1beta, IL-6, IL-8 and TNF-alpha was studied in short-time culture of separated stromal and epithelial cells. The cytokine secretion into culture medium was analyzed using immunoassay to evaluate the cytokine protein levels and bioassay to assess the bioactivity of the cytokines. Tissue samples of endometrium and ovarian endometriomas were obtained from 4 patients operated on for clinical reasons. Only IL-8 was found in all samples. IL-1beta and TNF-alpha were detected in the culture medium from most stromal cell samples, but in fewer media from epithelial cell samples. IL-6 was measurable in a few medium samples. Few of the samples displayed a bioactivity. There was no obvious difference between endometrium and endometriotic cell samples besides the production of IL-8 that seems to be lower in endometriotic tissue.

Topics: Adult; Cells, Cultured; Culture Media, Conditioned; Endometriosis; Endometrium; Epithelial Cells; Female; Humans; Immunoassay; Interleukin-1; Interleukin-6; Interleukin-8; Middle Aged; Stromal Cells; Tumor Necrosis Factor-alpha

To investigate the role of cytokines in peritoneal fluid on pathogenesis of endometriosis (EM).. Interleukin-6 (IL-6), interleukin-8(IL-8) and transforming growth factor-beta 1 (TGF-beta 1) contents in peritoneal fluid (PF) of 31 cases with EM were detected by enzyme linked immunoabsorbent assay (ELISA) and compared with the counterparts of 22 cases without EM (controls). The correlation analyses between cytokine concentrations in peritoneal fluid of EM patients and the severity of EM or dysmenorrhea score were performed.. The peritoneal fluid from patients with EM contained significantly greater amounts of IL-6 [(1.8 +/- 0.4) ng/L] and IL-8 [(1.7 +/- 0.5) ng/L] than those in controls [(1.2 +/- 0.2) ng/L and (1.4 +/- 0.3) ng/L respectively, P < 0.05]. However, in the amounts of TGF-beta 1 there were no significant difference (P > 0.05) between the two groups. The highest PF IL-6 and IL-8 concentrations were found in stage II, III and stag I, II EM respectively. A significant correlation between PF IL-6 content and the severity of disease was noted but there were no evidences of a relationship between concentrations of IL-8 and TGF-beta 1 and the severity of EM as well as between concentrations of three cytokines and dysmenorrhea score.. Unusual levels of IL-6 and IL-8 in PF of EM patients partly account for imbalance of the immunologically dynamic environment in peritoneal cavity of EM patients.

Topics: Adult; Ascitic Fluid; Endometriosis; Female; Humans; Interleukin-6; Interleukin-8; Transforming Growth Factor beta

To investigate the IL-8 and IL-10 levels in endometriosis and the regulated role of Neiyifang (NYF) on them.. Animal models with endometriosis after Cummings method was established. IL-8 and IL-10 levels were determined in using double antibody sandwich ELISA.. The IL-8 levels of serum and peritoneal fluid, and peritoneal macrophage amounts were markedly higher in the untreated group than those in the control group. The weights of endometriotic tissue and peritoneal macrophage amounts were markedly lower in the treated group of NYF, compared with the untreated group. The lipopolysaccharide stimulated IL-10 secretion in peritoneal macrophages from the untreated group were significantly lower compared with the control group and the treated group of NYF.. The abnormality of IL-8 and IL-10 levels in endometriosis may be related to the pathogenesis of the disease. The therapeutical mechanism of NYF was mediated by promotion of IL-10 secretion, and further restrained the composition of inflammatory media and the growth of endometriotic tissue.

Topics: Animals; Drugs, Chinese Herbal; Endometriosis; Female; Interleukin-10; Interleukin-8; Random Allocation; Rats; Rats, Sprague-Dawley

Peritoneal fluid in women with endometriosis contains an increased number of activated macrophages that secrete a variety of cytokines, including interleukin (IL)-6, IL-8, vascular endothelial growth factor, and tumor necrosis factor-alpha (TNF-alpha). Cytokines may be involved in the control of implantation and the growth of endometrial cells outside the uterus. In addition, several cytokines have been implicated in or directly associated with angiogenic activity in endometriosis. There could be a relationship between the levels of cytokines in the peritoneal fluid of patients with endometriosis and the status of the lesions in such patients. Peritoneal endometriosis can be classified as having red, black, or white lesions. Red lesions are known to be an active form of early endometriosis, because vascularization and mitotic activity are shown to be most prominent in these lesions. We found that the peritoneal fluid levels of TNF-alpha and IL-8 were significantly higher in patients with endometriosis, and correlated with the size and number of active lesions. In addition, TNF-alpha and IL-8 stimulated the growth of ectopic endometrial stromal cells. These cytokines with angiogenic activity may therefore have significant roles in the pathogenesis of endometriosis.

Topics: Cell Division; Cytokines; Disease Progression; Endometriosis; Endometrium; Extracellular Space; Female; Humans; Interleukin-8; Laparoscopy; Peritoneum; Severity of Illness Index; Stromal Cells; Tumor Necrosis Factor-alpha

To study the changes and significance of interleukin-6 (IL-6), interleukin-8(IL-8), tumor necrosis factor-alpha (TNF-alpha) and nitric oxide (NO) levels in vitro cultured system of peripheral mononuclear cells in patient with endometriosis(Em).. Peripheral blood mononuclear cells from 31 cases of endometriosis, classified into mild group (stage I and II, n = 11), and severe group (stage III and IV, n = 20) according to the Revised American Fertility society criteria 1985, and 15 normal controls were separated and induced by lipopolysaccharide and phytohemagglutinin, followed by 48 hour incubation in vitro, then the supernatant were collected. The levels of IL-6, IL-8 and TNF-alpha were measured by enzyme linked immunosorbent assays, the concentrations of NO, expressed by nitrite and nitrate (NO2-/NO3-) content were measured by Griess methods.. The concentrations of IL-6, IL-8, TNF-alpha and NO were significantly higher in both mild and severe Em groups than those in controls (P < 0.01). No differences between the mild and severe Em groups was seen. There were strong positive correlation between the levels of IL-6 and IL-8, TNF-alpha and NO in patients with endometriosis (P < 0.01).. The immune function of peripheral mononuclear cell in patients with endometriosis was disordered. It may activate peripheral mononuclear cells to produce high levels of IL-6, IL-8, TNF-alpha and NO, which may take part in the pathogenesis of endometriosis.

Topics: Endometriosis; Female; Humans; Interleukin-6; Interleukin-8; Leukocytes, Mononuclear; Nitric Oxide; Tumor Necrosis Factor-alpha

To investigate the role of interleukin-8 (IL-8) in peritoneal fluid of patients with endometriosis in the pathogenesis of endometriosis.. Peritoneal fluid was collected by laparoscopy. Endometrial and endometriotic stromal cells were obtained from normal endometrium and from chocolate cyst linings of the ovary.. Department of Obstetrics and Gynecology of Tottori University Hospital, Yonago, Japan.. Forty women who underwent either laparoscopy or laparoscopic surgery.. The peritoneal fluid concentration of IL-8 was analyzed by enzyme-linked immunosorbent assay, and the correlation between the IL-8 concentration and the extent of active endometriosis was investigated. The effect of IL-8 on cell proliferation was examined by tetrazolium bromide and thymidine incorporation. The expression of IL-8 receptor was examined in stromal cells by reverse transcription polymerase chain reaction.. The level of IL-8 in peritoneal fluid was significantly higher in patients with endometriosis than in patients without endometriosis. A significant correlation was noted with the extent of active endometriosis. Interleukin-8 significantly increased the number of cells and DNA synthesis in the endometrial and endometriotic stromal cells in a dose-dependent manner. Transcripts of IL-8 receptor type A were detected in stromal cells.. The present study suggests that IL-8 found in the peritoneal fluid of patients with endometriosis contributes to the pathogenesis of endometriosis.

Topics: Antigens, CD; Ascitic Fluid; Cell Division; Cells, Cultured; Endometriosis; Female; Humans; Interleukin-8; Receptors, Interleukin; Receptors, Interleukin-8A; Stromal Cells

There is increasing evidence that immunological mechanisms play a role in the pathogenesis and pathophysiology of endometriosis. It was therefore of interest to study interleukin-8 (IL-8), a chemokine, in the peritoneal fluid and peripheral blood of women undergoing laparoscopic procedures. The presence and concentrations of IL-8 in relation to endometriosis, infertility and abdominal pain were evaluated. Samples of peritoneal fluid (n = 49) and peripheral blood (n = 50) were obtained from 50 consecutive patients undergoing laparoscopic surgery for various gynaecological indications (abdominal pain, infertility, sterilization). IL-8 was present in the peritoneal fluid of most women (87%). The concentration of IL-8 in the peritoneal fluid was higher in women with endometriosis compared to women without (P = 0.02). This difference was more pronounced in early (stage 1) endometriosis (P = 0.001). IL-8 concentrations in the peritoneal fluid were also higher in women with early endometriosis compared to women with later stages of the disease (P = 0.003). Peripheral blood concentrations did not correlate with peritoneal fluid concentrations of IL-8 and/or the presence of endometriosis. We conclude that IL-8 is an important factor that may contribute to the pathogenesis of endometriosis possibly by promoting neovascularization. This information can be a guide in the development of new therapeutic approaches for the treatment of endometriosis.

Topics: Abdominal Pain; Adult; Ascitic Fluid; Endometriosis; Female; Follicular Phase; Humans; Infertility, Female; Interleukin-8; Laparoscopy; Luteal Phase; Menstrual Cycle

To evaluate basal (constitutive) and stimulated synthesis of tumor necrosis factor alpha (TNF alpha), interleukin (IL)-8, IL-10 by peritoneal macrophages (PM) in women with endometriosis.. Peritoneal macrophages were cultured in the presence or absence of lipopolysaccharide (LPS) for 24 hours. Peritoneal fluids (PF) and PM supernatants were assayed for cytokines using ELISA.. Institute for the Study and Treatment of Endometriosis and university-based research laboratories.. Fertile controls undergoing tubal ligation (n = 8) and women with endometriosis (n = 17).. Peritoneal fluid samples were obtained at the time of diagnostic laparoscopy (endometriosis group) or laparoscopy for tubal ligation; both were performed in the midluteal phase of the cycle.. Both basal and LPS stimulated production of TNF alpha, IL-8, and IL-10 by the PM were elevated significantly in women with endometriosis as compared with the fertile controls.. This study demonstrated that cytokines TNF alpha, IL-8, and IL-10 are synthesized at greater than normal levels by basal and stimulated PM from women with endometriosis. The levels of TNF alpha and IL-8 correlated with the levels in the PF, suggesting that PM are the principal source of these cytokines in the PF.

Topics: Ascitic Fluid; Cells, Cultured; Cytokines; Endometriosis; Female; Humans; Interleukin-10; Interleukin-8; Lipopolysaccharides; Macrophages, Peritoneal; Tumor Necrosis Factor-alpha

To investigate the capacity of peripheral blood monocytes (PBM) from women with endometriosis to secrete tumor necrosis factor-alpha (TNF-alpha), interleukin (IL) IL-6, IL-8, and IL-10.. Peripheral blood monocytes were cultured in the presence and absence of lipopolysaccharide for 24 hours before assessment of cytokines in supernatants by enzyme immunoassay.. Institute for the Study and Treatment of Endometriosis and university-based research laboratories.. Fertile controls, n - 10; women with endometriosis, n = 20.. None.. Basal synthesis of TNF-alpha, IL-6, and IL-8 and the lipopolysaccharide-stimulated synthesis of TNF-alpha by PBM from women with endometriosis was significantly greater in comparison to that of fertile controls. Endometriosis had no effect on the basal or stimulated synthesis of IL-10.. These data show that endometriosis is associated with increased basal and stimulated synthesis and secretion of several different cytokines by PBM. Each of the cytokines found to be affected has the capacity to play a role in the symptomatology or pathogenesis of the disease.

Topics: Cytokines; Endometriosis; Female; Humans; Interleukin-10; Interleukin-6; Interleukin-8; Lipopolysaccharides; Monocytes; Tumor Necrosis Factor-alpha

Interleukin-8 (IL-8) is a chemoattractant and activating factor for human neutrophlls and a potent angiogenic agent. The peritoneal fluid of women with endometriosis has been shown to have increased neutrophil chemotactic activity. We postulate that IL-8 may be an important modulator in the pathogenesis of endometriosis and adhesion formation. We first investigated IL-8 concentrations in the peritoneal fluid of women with or without endometriosis, then assessed peritoneal mesothelial cells as a potential source of peritoneal fluid IL-8. Northern blot analysis and enzyme-linked immunosorbent assay (ELISA) were used to investigate IL-8 mRNA and protein modulation. The mean concentration of IL-8 in samples obtained from control patients (n = 28) was 4.8 +/- 0.5 pg/ml; from patients with minimal-mild endometriosis (n = 24) was 27.5 +/- 2.6 pg/ml; and from patients with moderate-severe endometriosis (n = 21) was 530.2 +/- 65.1 pg/ml. Confluent mesothelial cells were incubated with human recombinant IL-1 alpha (0.01-100 IU/ml) or tumour necrosis factor (TNF)-alpha (0.01 to 100 ng/ml) for 2-24 h. IL-8 mRNA was detectable in non-treated cells, however both IL-1 alpha and TNF-alpha induced higher amounts of IL-8 mRNA in a dose- and time-dependent manner. Non-treated mesothelial cells in culture also produced and secreted IL-8 protein quantified by ELISA, but again higher concentrations were induced by IL-1 alpha and TNF-alpha treatment. In conclusion, we found that IL-8 concentrations were elevated in peritoneal fluids from women with endometriosis. Cultured mesothelial cells expressed cytokine-inducible IL-8 mRNA and secreted IL-8 protein. The regulated expression of this angiogenic factor may play a role in pathogenesis of endometriosis.

Topics: Ascitic Fluid; Blotting, Northern; Cells, Cultured; Endometriosis; Epithelium; Female; Gene Expression; Humans; Immunoassay; Immunohistochemistry; Interleukin-8; RNA, Messenger; Statistics, Nonparametric

To investigate the presence of interleukin-8 (IL-8), a macrophage-derived angiogenic factor, in peritoneal fluid (PF) of women with and without endometriosis.. Case-control study.. University hospital.. Eighteen women with laparoscopic findings of mild to severe endometriosis, and nine women with no visual evidence of pelvic pathology.. Peritoneal fluid IL-8 levels were determined using an ELISA. Interleukin-8 concentrations were compared among women with and without endometriosis. Correlation between PF IL-8 concentration and endometriosis stage was investigated.. Interleukin-8 was detectable in the PF of a majority of women (67%). Interleukin-8 concentrations were higher in the PF of women with endometriosis than in matched normal controls. A significant correlation between PF IL-8 concentration and endometriosis stage was noted.. We hypothesize that IL-8 is an important angiogenic factor that contributes to the pathogenesis of endometriosis by promoting the neovascularization of ectopic endometrial implants.

Topics: Adult; Ascitic Fluid; Case-Control Studies; Endometriosis; Enzyme-Linked Immunosorbent Assay; Female; Humans; Interleukin-8; Osmolar Concentration; Reference Values