Soluble c-Met expression in the peritoneal fluid and serum of patients with different stages of endometriosis

Introduction

Hepatocyte growth factor (HGF), also known as scatter factor, and its receptor c-Met have been shown to be implicated in endometriosis. HGF acts as a mitogen, motogen, and morphogen on endometrial epithelial cells. The expression of c-Met on human endometrial cells has been reported. Many proteins are proteolytically released from the surface by a process known as ectodomain shedding. The aim of this study was to determine the levels of soluble c-Met (s-cMet) in the peritoneal fluid (PF) and serum samples of patients with different stages of endometriosis.

Material and methods

39 PF and serum samples from normal healthy and 130 samples from different stages of patients with endometriosis (33 cases of stage I, 38 stage II, 30 stage III and 29 stage IV) were included in this study. Total protein concentration (TPC) and the level of s-cMet in the PF and serum were determined by Bio-Rad protein assay based on the Bradford dye procedure and enzyme-linked immunosorbent assay, respectively.

Results

No significant change in the TPC was seen in the serum and PF of patients with endometriosis when compared with normal controls. Results obtained demonstrated that all PF and serum samples presented s-cMet expression, whereas, starting from stages I to IV endometriosis, a significant increase of s-cMet expression was observed as compared to controls.

Conclusion

The results of this study show that a high expression of s-cMet is correlated with advanced stages of endometriosis. It is also concluded that the detection of serum and PF s-cMet may be useful in classifying endometriosis.     

High soluble CD44 concentration in peritoneal fluid in endometriosis

The concentration of the soluble form of CD44, a competitive inhibitor for CD44 and hyaluronic acid binding, is higher in the peritoneal fluid of patients with endometriosis than in the peritoneal fluid of patients without endometriosis.     

Correlation between serum and peritoneal fluid glutathione S-transferases T1 concentration with different stages of endometriosis

Endometriosis is a gynecological disease defined by the histological presence of endometrial glands and stroma outside the uterine cavity. Ectopic endometrial cell proliferation and chronic inflammation in endometriosis were shown to be associated with oxidative stress (OS) induction. OS is a condition in which reactive oxygen species (ROS) overproduction and antioxidant deficiency cause a shift in oxidant/antioxidant balance. Glutathione S-transferases (GSTs) comprise a family of eukaryotic and prokaryotic phase II metabolic isozymes best known for their ability to catalyze the conjugation of the reduced form of glutathione (GSH) to xenobiotic substrates for the purpose of detoxification. The aim of this project was to study the concentrations of GSTT1 in the serum and peritoneal fluid (PF) of patients with different stages of endometriosis. Frothy two PF and serum from normal and 152 from different stages of patients with endometriosis (stage I: n = 30, stage II: n = 39, stage III: n = 43 and stage IV: n = 40) were included in this study. The level of GSTT1 in the serum was determined by enzyme linked immunosorbent assay (ELISA). The results showed the presence of GSTT1 in all serum and peritoneal fluid samples, while, starting from stages I to IV endometriosis, a significant decrease in GSTT1 concentration was seen as compared to controls. It is concluded that levels of GSTT1 is negatively correlated with advanced stages of endometriosis. It is also suggested that the detection of serum and/or peritoneal fluid GSTT1 concentration may be valuable in the classifying of endometriosis.     

Expression of inflammatory cytokines in serum and peritoneal fluid from patients with different stages of endometriosis

Objective: To investigate the level of inflammatory cytokines in endometriosis patients, and explore the relationship between IL-37 concentration and endometriosis stages.

Methods: Inflammatory cytokine concentrations from 27 patients with different stages of endometriosis and 52 controls without endometriosis were examined by ELISA. Then, the specificity and sensitivity of cytokines for distinguishing from controls and the different stages of endometriosis were analyzed using the ROC curve.

Results: The difference in serum concentrations of IL-37, IL-17A, IL-10, and IL-2 between the endometriosis and control groups was statistically significant (p?p?p?=?.0034), IL-10 concentrations in PF were significantly lower in the early-stages of endometriosis than in the more advanced groups (p?=?.0439), and IL-4 concentration in PF was significantly higher in more advanced endometriosis (p?=?.0228). The sensitivity and specificity of serum IL-37 for distinguishing endometriosis were 81.48% and 83.33%, respectively, and the cutoff concentration was 69.84?pg/ml. For IL-17A, the sensitivity and specificity were 96.30% and 100%, respectively, and the cutoff concentration was 57.54?pg/ml. For IL-10, the sensitivity and specificity was 92.59% and 100%, respectively, and the cutoff concentration was 3.301?pg/ml. For IL-2, the sensitivity and specificity were 74.07% and 93.75%, respectively, and the cutoff concentration was 1.813?pg/ml. For PF IL-2, the sensitivity and specificity were 29.73% and 100%, respectively, and the cutoff concentration was 1.06?pg/ml.

Conclusions: IL-37, IL-17A, IL-10, and IL-2 may play a significant role in immune response in endometriosis. IL-37 levels may be used as a diagnostic marker for endometriosis.     

VEGF concentration in peritoneal fluid of patients with endometriosis

The theory of Sampson that endometrial cells and fragments desquamated during the menstrual period are transported through fallopian tubes into the peritoneal cavity where they implant, proliferate and develop into endometriotic lesions is generally accepted. There is increasing evidence that immunological mechanisms play a role in the pathogenesis and pathophysiology of endometriosis. Excessive endometrial angiogenesis is proposed as an important mechanism in the pathogenesis of endometriosis. Evidence is reviewed for the hypothesis that the endometrium of women with endometriosis has an increased capacity to proliferate, implant and grow in the peritoneal cavity. From the known angiogenic factors, vascular endothelial growth factor (VEGF) has emerged as a pivotally important regulator of normal angiogenesis and pathological neovascularization. In present study we evaluated the concentrations of VEGF in peritoneal fluid of patients with endometriosis and showed no correlation between AFS score and VEGF concentration in peritoneal and in ovarian endometriosis. Above results do not confirm former observations indicating the role of VEGF in endometriosis pathogenesis.     

Soluble HLA-G in the peritoneal fluid of women with endometriosis

The aim of this study was to determine the level of soluble HLA-G molecules in the peritoneal fluid of endometriosis patients. The findings demonstrate that a soluble HLA-G level in the peritoneal fluid of women with endometriosis is similar to that of the control group.     

CA 125 in peritoneal fluid and serum of patients with endometriosis

Serum and peritoneal fluid collected from 48 patients at laparoscopy was assayed for CA 125. Geometric mean serum CA 125 was similar in 21 patients with predominantly early stage endometriosis (13.4 U/ml; confidence intervals (C.I.) 10.8-16.7) and 27 patients without endometriosis (12.9 U/ml; C.I. 9.7-17.2). Mean peritoneal fluid CA 125 was higher in the endometriosis patients (mean 44.4 U/ml; C.I. 34.5-57.2) than controls (35.7 U/ml; C.I. 30.0-42.2), but the difference was not significant. Peritoneal fluid volume was increased in endometriosis patients (p = 0.013), and only after correction for peritoneal fluid volume did total antigenic concentration in aspirated peritoneal fluid show significant elevation (p = 0.0015) in endometriosis patients (mean 340 U/ml; C.I. 212-547) compared with controls (125 U/ml; C.I. 85-184). These results suggest that although CA 125 may be expressed in peritoneal fluid in endometriosis, it is unlikely to be of use as a marker of disease activity in peritoneal fluid obtained by culdocentesis.     

Soluble intercellular adhesion molecule 1 in the peritoneal fluid of patients with endometriosis correlates with the extension of peritoneal implants

OBJECTIVE: To investigate the correlation of the concentrations of soluble intercellular adhesion molecule 1 (sICAM-1) in the peritoneal fluid (PF) with the extent of peritoneal endometriotic lesions and with the presence of red lesions (RLs). STUDY DESIGN: Sixty-seven patients with endometriosis and 19 controls with normal pelvis had PF samples collected during laparoscopy. RESULTS: The concentrations of sICAM-1 in the PF of patients and controls were not different. Patients with peritoneal implant scores 4 or more had higher concentrations of sICAM-1 in the PF than those with implant scores less than 4 (P=0.018) and controls (P=0.031). No significant difference was found in sICAM-1 levels in patients with and without RLs. No correlation was detected between sICAM-1 and interleukin 1beta (IL-1beta) in either patients or controls. CONCLUSIONS: The increased concentrations of sICAM-1 in the PF of patients with greater implant scores may indicate an active shedding of the molecule from the endometriotic peritoneal tissue. No significant association was found with the presence of RLs. The levels of IL-1beta in the PF were not a relevant factor influencing the concentrations of sICAM-1.     

Endometrial antibodies in serum and peritoneal fluid of infertile patients with and without endometriosis

Passive hemagglutination assay was used to evaluate endometrial antibodies in serum and peritoneal fluid of 37 patients with endometriosis and 54 patients without endometriosis. The results showed that the concentration of antibody titers in serum and peritoneal fluid was significantly higher for endometriosis than control patients. The severity of endometriosis has no effect on antibody concentration. Furthermore, the concentration of endometrial antibody titers was significantly higher in serum than peritoneal fluid of patients with endometriosis. These results suggest that serum endometrial antibody assay is specific and valuable for the diagnosis and progress of endometriosis.     

E-cadherin in the serum and the peritoneal fluid of women with endometriosis

Endometriosis, a disease of unclear etiopathogenesis, is a quite common problem in gynecology. It is thought that the retrograde flow of the menstrual debris to the peritoneal cavity plays an important role in the origin of endometriosis but the mechanism of endometrial cells implantation remains unknown. Recently many centers have reported the importance of adhesion molecules in this process. We studied the concentrations of E-cadherin in the serum and the peritoneal fluid of women with endometriosis. Our results show a significant decrease of E-cadherin concentrations in sera and peritoneal fluids in women with endometriosis. We suggest that screening the level of E-caherin may be useful in the monitoring the therapy of endometriosis.     

Increased levels of biglycan in endometriomas and peritoneal fluid samples from ovarian endometriosis patients

In our previous low-density-array gene-expression analysis we found an increased expression of biglycan gene in ovarian endometriosis patients. In the present study we evaluated biglycan expression at the protein level in tissue, serum and peritoneal fluid (PF) from ovarian endometriosis patients, patients with benign ovarian cysts and healthy women. Twenty samples of endometriomas and 27 of control tissues (benign ovarian cysts and eutopic endometrium of healthy women) were obtained laparoscopically or by curettage. Serum and PF samples were collected from 56 ovarian endometriosis patients and 40 controls (patients with benign cysts and healthy women). Tissue biglycan levels and serum and PF biglycan concentrations were determined by Western blotting and ELISA, respectively. Biglycan was detected in endometriomas and in benign cysts tissues but differed in glycosylation levels. The PF biglycan concentrations were significantly increased in ovarian endometriosis patients (mean?±?SD?=?220.3?±?190.5?pg/mg protein) compared to the whole control group (101.9?±?94.7?pg/mg protein, p?相似文献   

Production of fibronectin by peritoneal macrophages and concentration of fibronectin in peritoneal fluid from patients with or without endometriosis

Fibronectin, a known growth factor for fibroblasts, is produced by alveolar macrophages from patients with interstitial pulmonary fibrosis. Because peritoneal macrophages have been implicated in the disease process of endometriosis, we measured the production of fibronectin by peritoneal macrophages in vitro and the concentration of fibronectin in peritoneal fluid samples. Twenty-nine patients had a normal pelvis, 22 had endometriosis, and 14 had tubal occlusion and/or adhesions. Human peritoneal macrophages demonstrated de novo synthesis of fibronectin. The peritoneal macrophage fibronectin was detected by an enzyme-linked immunosorbent assay for serum fibronectin. Peritoneal macrophages from patients with endometriosis produced approximately three times the amount of fibronectin as normal patients or patients with tubal occlusion and/or adhesions (P less than or equal to .01 and P less than or equal to .02, respectively). The mean peritoneal fluid concentration of fibronectin, however, was about 30% lower in patients with endometriosis than in normal patients (P less than or equal to .02). We suggest that increased peritoneal macrophage fibronectin production in patients with endometriosis may contribute to the adhesion formation and associated reactive fibrosis seen in this disease, and may also influence the implantation of endometrial cells and their subsequent growth in the pelvis.     

Low serum and peritoneal fluid concentration of interferon-gamma-induced protein-10 (CXCL10) in women with endometriosis

OBJECTIVE: To evaluate serum and peritoneal fluid concentrations of interferon-gamma-inducible protein-10 (CXCL10), a chemokine involved in local immune function, in women with endometriosis. DESIGN: Prospective study. SETTING: Division of Obstetrics and Gynecology, University of Siena. PATIENT(S): A total of 147 women were divided in two groups: women with (n = 77) and without (n = 70) endometriosis. INTERVENTION(S): Serum and peritoneal fluid were collected from all patients undergoing laparoscopy. MAIN OUTCOME MEASURE(S): CXCL10 concentrations were measured by a specific ELISA. RESULT(S): Serum CXCL10 concentrations in women with endometriosis were significantly lower than in those without endometriosis. No statistically significant difference between women with early endometriosis and those with advanced endometriosis was found. CXCL10 concentrations in peritoneal fluid of women with advanced endometriosis were significantly lower than in that of women with an early stage of, or without, endometriosis. CONCLUSION(S): The decreased concentrations of CXCL10 in serum and peritoneal fluid of women with endometriosis indicate an impaired immune activity in women with endometriosis.     

Expression of cadherins and CD44 isoforms in human endometrium and peritoneal endometriosis

BACKGROUND: To evaluate (a) cadherins and CD44 expression in normal endometrium and in peritoneal endometriosis, and (b) to correlate, their expressions with clinicopathological parameters. METHODS: E- and N-cadherin, CD44 isoforms, CD44v3 and CD44v6 expressions were evaluated: (a) by immunoblotting in endometrium (n = 6) and in peritoneal endometriotic samples (n = 7) and, (b) by immunohistochemistry in endometrium (n = 15) and in peritoneal endometriotic samples (n = 23). RESULTS: By immunoblotting, endometrium expressed E- and N-cadherin, CD44 isoforms, CD44v3 and CD44v6. Similar results were observed in peritoneal endometriosis. By immunohistochemistry, in endometrium, E-cadherin was restricted to epithelial cells. Its expression remained constant throughout the menstrual cycle. N-cadherin was detected in both epithelial and stromal cells in the proliferative phase but was restricted to epithelial cells in the secretory phase. CD44 immunostaining was detected in the secretory but not in the proliferative phase. Decreased expression of E-cadherin (p < 0.01) and CD44 (p < 0.01) in epithelial cells was found in peritoneal endometriosis as compared with normal endometrium. In endometriotic stromal cells, decreased CD44 expression was found. In peritoneal endometriosis, we observed a decreased expression of E-cadherin in advanced stages of the disease (p < 0.01). CONCLUSION: Our results suggest that E-cadherin and CD44 proteins could be involved in the development of endometriotic lesions. Investigation of the mechanisms of altered adhesion molecule expression may contribute to the understanding of the behavior of endometriotic cells.     

Behaviour of cytokine levels in serum and peritoneal fluid of women with endometriosis

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 observed changes may be due both to the increased peritoneal macrophage activity and to the larger recruitment of PBMC and autocrine release by endometriotic cells.     

Decreased levels of interleukin-18 in peritoneal fluid but not in serum of patients with endometriosis

OBJECTIVE: To determine the concentrations of interleukin-18 (IL-18) in peritoneal fluid and serum in patients with endometriosis in comparison with the control group. DESIGN: A prospective analytical study. SETTING: The obstetrics and gynecology department of an academic training hospital. PATIENT(S): Forty-four patients who underwent laparoscopic surgery for benign gynecologic diseases. INTERVENTION(S): Specimens of peripheral blood and peritoneal fluid were obtained before and during laparoscopic procedures, and the levels of IL-18 were analyzed. MAIN OUTCOME MEASURE(S): The concentrations of IL-18 in peritoneal fluid and serum were correlated with the presence of endometriosis, disease stage, and the phase of the menstrual cycle. RESULT(S): Interleukin-18 was detectable in 98% of the peritoneal specimens and 84% of the serum specimens of the patients tested. Peritoneal fluid IL-18 concentrations were statistically significantly lower in patients with endometriosis than in patients without endometriosis; the difference in serum IL-18 levels showed no statistically significant difference between the patients with and without endometriosis. The concentrations of IL-18 in peritoneal fluid and serum were not correlated with the stage of endometriosis or the phase of the menstrual cycle. CONCLUSION(S): Our results suggest that the decreased levels of IL-18 in peritoneal fluid in patients with endometriosis as compared with the control group may play an important role in the pathogenesis of this disease.     

Antibodies to Chlamydia trachomatis in serum and peritoneal fluid of women with endometriosis

The specifics of inflammation created by infection with Chlamydia trachomatis could be favourable to the genesis of endometriosis. To investigate this hypothesis, we studied the association between Chlamydia trachomatis specific IgG and IgA antibodies in serum and the peritoneal fluid of 51 women undergoing laparoscopic surgery. There was no significant difference between women with and without endometriosis with respect to the incidence of IgG and IgA in serum or the peritoneal fluid. The results of our preliminary study did not show any significant link between past infection with Chlamydia trachomatis and the presence of endometriosis.     

Elevated glycodelin-A concentrations in serum and peritoneal fluid of women with ovarian endometriosis

Abstract

The aim of this study was to evaluate serum and peritoneal fluid (PF) glycodelin-A concentrations in women with ovarian endometriosis. Ninety-nine matched pairs of serum and PF samples were included in our study. The case group comprised 57 women with ovarian endometriosis and the control group 42 healthy women undergoing sterilization or patients with benign ovarian cysts. Glycodelin-A concentrations were measured using ELISA. Endometriosis patients had significantly higher serum and PF glycodelin-A concentrations compared to controls, and this increase was observed in both proliferative and secretory cycle phases. Glycodelin-A concentrations were more than 10-fold higher in PF than in serum and correlated with each other. Intensity and frequency of menstrual pain positively correlated with glycodelin-A concentrations. Sensitivity and specificity of glycodelin-A as a biomarker for ovarian endometriosis were 82.1% and 78.4% in serum, and 79.7% and 77.5% in PF, respectively. These results indicate that Glycodelin-A has a potential role as a biomarker to be used in combination with other, independent marker molecules.