Effect Of Peritoneal Fluid Research Articles

Pain Mechanisms in Peritoneal Diseases Might Be Partially Regulated by Estrogen.

To identify factors influencing the differential pain pathogenesis in peritoneal endometriosis (pEM) and peritoneal carcinomatosis in ovarian cancer (pOC), we undertook an experimental study. Tissue samples of 18 patients with pEM, 15 patients with pOC, and 15 unaffected peritoneums as controls were collected during laparoscopy or laparotomy. Immunohistochemical stainings were conducted to identify nerve fibers and neurotrophins in the tissue samples. Additionally, 23 pEM fluids, 25 pOC ascites fluids, and 20 peritoneal fluids of patients with myoma uteri as controls were collected. In these fluids, the expression of neurotrophins was evaluated. The effects of peritoneal fluids and ascites on the neurite outgrowth of chicken sensory ganglia were estimated by using a neuronal growth assay. An electrochemiluminescence immunoassay was carried out to determine the expression of estrogen in the peritoneal fluids and ascites. The total and sensory nerve fiber density was significantly higher in pEM than in pOC ( P < .001 and P < .01). All neurotrophins tested were present in tissue and fluid samples of pEM and pOC. Furthermore, the neurotrophic properties of pEM and pOC fluids were demonstrated, leading to sensory nerve fiber outgrowth. Estrogen concentration in the peritoneal fluids of pEM was significantly higher compared to ascites of pOC ( P < .001). The total and sensory nerve fiber density in the tissue samples as well as the estrogen expression in the peritoneal fluid of pEM was considerably higher than that in pOC, representing the most notable difference found in both diseases. This might explain the differential pain perception in pEM and pOC. Therefore, estrogen might be a key factor in influencing the genesis of pain in endometriosis.

The Effect of Peritoneal Fluid from Patients with Endometriosis on Mitochondrial Function and Development of Early Mouse Embryos

BackgroundPeritoneal fluid (PF) from patients with endometriosis can inhibit early embryo development via probable functional changes of embryo mitochondria in the early stage of embryo development. The purpose of this study was to determine the effect of PF from patients with endometriosis on mitochondrial function and development of early mouse embryos.Methodology/Principal FindingsPF was collected from patients with infertility and endometriosis, infertility due to tubal factors, and normal control subjects, and the level of NO was measured. Early murine embryos were then cultured with PF from normal control subjects, those with endometriosis, and with human tubal fluid (HTF), respectively. Cleavage and blastulation rates, mitochondrial DNA (mtDNA) copy numbers, adenosine triphosphate (ATP) level, and mitochondrial membrane potential (ΔΨm) of the different groups were compared. The NO level in the PF of patients with endometriosis was significantly greater than in those without endometriosis and control patients. The embryos cultures with PF from patients with endometriosis had a lower cleavage rate and blastulation rate, and higher ATP and ΔΨm level at the 2- and 4-cell stages. No significant difference was found in mtDNA copies among the 3 groups.Conclusions/SignificancePF from patients with endometriosis can inhibit early embryo development via probable functional changes of embryo mitochondria in the early stage of embryo development. Understanding the effects of PF on embryo development may assist in developing new methods of treatment for infertility.

Overexpression of nerve growth factor in peritoneal fluid from women with endometriosis may promote neurite outgrowth in endometriotic lesions

To investigate the role of the nerve growth factor (NGF) in the endometriosis-associated innervation in the development of endometriosis-associated symptoms, 41 peritoneal fluid samples (PF) from patients with surgically and histologically proven endometriosis and 20 PF from patients with other gynecologic conditions were analyzed with Western blot and a novel in vitro model using dorsal root ganglia (DRG) to show neuronal outgrowth; endometrial cells also were analyzed. The results suggest that the PF of endometriosis patients and endometriotic lesions have neurotropic properties, because the Western blot analysis and the cell culture stainings showed NGF expression, and the neurite outgrowth of DRG treated with PF of patients with endometriosis was significantly higher than when treated with PF of patients without endometriosis. Furthermore, blocking NGF with both anti-NGF and K252a leads to a significant decrease in neurite outgrowth.

Effect of peritoneal fluid from endometriosis patients on neuroblastoma cells in culture

Aim. Endometriosis is often associated with lower abdominal pain, dysmenorrhea, dyspareunia, and chronic pelvic pain. There is no correlation between the extent of endometriosis and the intensity of pain. The mechanism of pain in endometriosis is unknown. The aim of our study was to investigate the influence of peritoneal fluid (PF) from endometriosis patients on cultured neural cells that are the morphological basis of nociception, and to determine whether there was a relationship between the rAFS staging and an elevation of TGF-β1 production by these cells.Methods. Different human neuroblastoma cell lines were grown to 3/4 confluence and then cultured in presence of PF pooled according to the presence of no, mild, or severe endometriosis. After 6 and 24 h of incubation, the morphological changes were assessed and the metabolic activity was determined.Results. The different cell lines showed strongly varying proliferation and aggregation patterns. The metabolic activity was also varying between cell lines, but no consistently increased cell turnover in the PF when compared with the control medium nor associated to a particular, endometriosis-derived PF pool could be shown. In this experimental setting, we have observed that the cell proliferation in the presence of PF was inhibited, and not enhanced as it might have been expected. Measurement of TGF-β1 showed higher production rates for this cytokine under exposure to PF than in controls for some but not all tested cell lines, but there was no association with the stage (rAFS) of the disease.Conclusion. The neuronal cell culture model may become a useful tool to investigate the endometriosis-derived pain, but different endpoints and cell lines may have to be introduced.

Effects of Peritoneal Fluid from Endometriosis Patients on the Release of Interferon-g-Induced Protein-10 and Interleukin-8 by Neutrophils and T Cells

Effects of Peritoneal Fluid from Endometriosis Patients on the Release of Interferon-g-Induced Protein-10 and Interleukin-8 by Neutrophils and T Cells

Effect of peritoneal fluid from endometriosis patients on neuroblastoma cells in culture

Effect of peritoneal fluid from endometriosis patients on neuroblastoma cells in culture

Correlation between soluble Fas level and apoptosis of T cells in ovarian carcinoma

Objectives The objectives were to examine the correlation between soluble Fas (sFas) level and apoptosis of T cells in peripheral blood and peritoneal fluid of patients with ovarian carcinoma and to investigate the possible sFas effect on T cell apoptosis. Study design Patients with stages I–II ovarian carcinoma ( n = 10) and patients with stages III–IV ovarian carcinoma ( n = 22), as well as ovarian benign tumors ( n = 8), were enrolled in the study. Apoptosis of and Fas expression on T cells from peripheral blood and peritoneal fluids were assessed by flow cytometry. Soluble Fas level was assayed using an ELISA kit. The effects of peritoneal fluid on Jurkat cell apoptosis with or without depletion of sFas were evaluated and compared in vitro. Results The sFas level and apoptosis of T cells in peripheral blood and peritoneal fluid from stages III–IV ovarian carcinoma were significantly higher than those from stages I–II ovarian carcinoma ( p < 0.01 in all instances) and benign ovarian tumor ( p < 0.01 in all instances). In peritoneal fluid, the sFas level and apoptosis of T cells from stages I–II ovarian carcinoma were significantly higher than those from benign ovarian tumor ( p < 0.01 in all instances), and the Fas expression on T cells from ovarian carcinoma were higher than those from benign ovarian tumor ( p < 0.05 in all instances). There was a positive correlation between the sFas level and the apoptosis of T cells in peritoneal fluids from stages III–IV ovarian carcinoma ( r = 0.647, p = 0.001). Peritoneal fluid of ovarian carcinoma could induce significant Jurkat cell apoptosis. The blocking of Fas expression on the Jurkat cell surface, but not the deletion of sFas, may remarkably restrain the apoptosis level. Conclusions Elevated sFas is correlated with apoptosis of T cells in peripheral blood and peritoneal fluid from ovarian carcinoma. Soluble Fas evidently does not affect T cell apoptosis, which is probably due to elevated Fas expression on T cells.

Effects of peritoneal fluid from endometriosis patients on the release of vascular endothelial growth factor by neutrophils and monocytes

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.

Study of Cytokine Profile and Angiogenic Potential of Peritoneal Fluid in Patients with External Genital Endometriosis

The concentrations of proinflammatory cytokines (IL-1beta, IL-6), vascular endothelium growth factor, tumor growth factor-beta, and insulin-like growth factor-1 were measured in the peritoneal fluid of patients with external genital endometriosis and healthy women by enzyme immunoassay. The effect of peritoneal fluid from patients with external genital endometriosis on proliferative activity of EA.Hy926 human endothelial cells was evaluated by the method based on the analysis of cell cycle by flow cytometry. The concentrations of IL-1beta, IL-6, and insulin-like growth factor-1 were increased in patients with endometriosis in comparison with healthy women. The peritoneal fluid from patients with endometriosis (but not from healthy women) significantly increased mitotic activity of endothelial cells and exhibited high angiogenic potential, which can promote implantation and growth of endometrial transplants. Presumably, insulin-like growth factor-1 stimulates this process.

003 Effects of peritoneal fluid from endometriosis patients on vascular endothelial growth factor release by neutrophils and monocytes

003 Effects of peritoneal fluid from endometriosis patients on vascular endothelial growth factor release by neutrophils and monocytes

IL-10-dependent down-regulation of MHC class II expression level on monocytes by peritoneal fluid from endometriosis patients

Endometriosis is a gynecologic disorder characterized by the ectopic growth of misplaced endometrial cells. Moreover, immunological abnormalities of cell-mediated and humoral immunity may be associated with the pathogenesis of endometriosis. The effects of peritoneal fluid (PF) from endometriosis patients on the expression levels of MHC class II and costimulatory molecules on the cell surfaces of monocytes were investigated. Compared to the PF of controls, the addition of 10% PF ( n = 10) from patients with endometriosis to culture medium significantly reduced the percentage of MHC class II-positive cells in cultures of a THP-1, monocytic cell line at 48 h. The effect of endometriosis patient PF (EPF) was dose-dependent, and similar effect was observed in peripheral blood monocytes. An inverse correlation was found between MHC class II expression level and IL-10 concentration in EPF ( r = − 0.518; p = 0.019) and in the supernatant of peripheral blood monocyte cultured in EPF ( r = − 0.459; p = 0.042) ( n = 20). The expression levels of costimulatory molecules (CD80 and CD86), but not of CD54 and B7-H1, were down-regulated by EPF. The mRNA level of HLA-DR was unaffected by EPF but protein level was reduced by EPF. Neutralizing IL-10 antibody abrogated MHC class II down-regulation on monocytes, which had been induced by EPF. However, in a functional assay, monocytes treated with EPF failed to stimulate T cell in mixed leukocyte reaction, although T cell proliferation was increased with EPF-treated monocytes and Staphylococcus enterotoxin B. These results suggest that MHC class II expression level on monocytes is down-regulated by EPF, but the cell stimulatory ability of monocytes does not coincide with MHC class II expression level.

Inhibitory effect of nitric oxide on the proliferation of endometrial cells

Objective: Development of endometriosis is associated with endometrial stromal cell proliferation. The effects of nitric oxide (NO) on proliferation of endometrial cells and the effects of peritoneal fluid from women with and without endometriosis on the production of NO and NO-induced changes were investigated.

Effect of peritoneal fluid from women with endometriosis on implantation in the mouse model

Objective: To investigate the potential role of peritoneal fluid (PF) from women with or without endometriosis in implantation in mice with use of the delayed implantation model. Design: A murine experimental model with markers of uterine receptivity and prospective comparison of the effects of human PF on implantation. Setting: Academic university and hospital program. Intervention(s): PF collected from women with and without endometriosis was injected intraperitoneally into recently mated mice. Main Outcome Measure(s): Implantation sites were counted in treated and untreated animals, and the αvβ3 integrin was measured in the pregnant mouse uterus by immunohistochemistry with in situ hybridization. Leukemia inhibitory factor and the β3 subunit of αvβ3 were measured by Northern blot during early pregnancy and after injections of PF. Result(s): Animals receiving PF from infertile women with endometriosis had a reduction in the number of implantation sites compared with animals that received PF from fertile women or from patients with recently treated endometriosis. In the mouse, expression of αvβ3 and leukemia inhibitory factor peaked at the time of implantation and was reduced by injections of human PF from infertile patients with endometriosis. Conclusion(s): Leukemia inhibitory factor and αvβ3 are coexpressed at the time of implantation in the mouse. PF from women with endometriosis has a detrimental effect on embryo implantation, perhaps by adversely affecting uterine receptivity.

Changes of cytokines levels in peritoneal fluids of patients with endometriosis and its effect on reproductive activity.

To study the changes of cytokines in peritoneal fluids of patients with endometriosis and their effects on reproductive activity, levels of tumor necrosis factor (TNF) and interleukin 6 (IL-6) in peritoneal fluids and peritoneal macrophages' culture supernatant were studied by using enzyme linked immunoassay (ELISA) in 14 infertile patients with endometriosis (EMT group) and 11 infertile women with normal pelvis (control group). The effects of peritoneal fluids in patients with endometriosis in vitro on sperm motility and development of 2-cell mouse embryos were also studied. The results showed that the levels of TNF and IL-6 in peritoneal fluids and peritoneal macrophages' supernatant in EMT group were elevated significantly as compared with those in the control group (P < 0.01). The percent of sperm straight line movement and total sperm motility were decreased significantly in EMT group (P < 0.01). The percent of 2-cell mouse embryos developing to 16-cells was 32.5% in EMT group, while 47.6% in control group after 48 h co-culture with peritoneal fluid (P < 0.01). It is likely that the elevation of peritoneal fluid cytokines in patients with endometriosis may play a role in infertility associated with endometriosis.

Effect of peritoneal fluid on whole body and segmental multiple frequency bioelectrical impedance in patients on peritoneal dialysis.

We investigated the ability of whole body and segmental multiple frequency bioelectrical impedance (MFBIA) to detect peritoneal fluid in peritoneal dialysis patients. Prospective study. Teaching hospital renal unit. Patients on regular peritoneal dialysis. Whole body and segmental MFBIA measurements before and after drainage of peritoneal fluid. Changes estimated by MFBIA in total body water (-0.4 (0.8) litres) and extracellular water (-0.3 (0.3) litres) were much lower than the actual changes (2.0 (0.4) litres), P<0.0001. Derived resistances Recf and Ricf increased significantly for the trunk but not for total body measurements and changes did not correlate with volumes of fluid drained. MFBIA is limited in its ability to detect intraperitoneal fluid, using both whole body and segmental techniques.

Peritoneal fluid modulates the sperm acrosomal exocytosis induced by N-acetylglucosaminyl neoglycoprotein.

The effect of peritoneal fluid (PF) on the human sperm acrosome reaction (AR) was tested. Sperm was pre-incubated with PF and the AR was induced by calcium ionophore A23187 and a neoglycoprotein bearing N-acetylglycosamine residues (NGP). The AR induced by calcium ionophore was inhibited 40% by PF from controls (PFc) and 50% by PF from the endometriosis (PFe) group, but not by PF from infertile patients without endometriosis (PFi). No significant differences were found in the spontaneous AR. When the AR was induced by NGP, pre-incubation with PFc reduced (60%) the percentage of AR, while PFe and PFi caused no significant differences. The average rates of acrosome reactions obtained in control. NGP- and ionophore-treated sperm showed that NGP-induced exocytosis differed significantly between the PFc (11%) and PFe/PFi groups (17%), and the ionophore-induced AR was higher for PFi (33%) than PFc/PFe (25%). The incidence of the NGP-induced AR was reduced in the first hour of pre-incubation with PFc and remained nearly constant throughout 4 h of incubation. The present data indicate that PF possesses a protective factor which prevents premature AR.

Suppression of macrophage activation by peritoneal fluid from patients with endometriosis.

To investigate the effects of peritoneal fluid from patients with endometriosis on mouse peritoneal macrophages (Mphi), peritoneal fluid from endometriosis patients (n = 15) were added to a monolayer of C3H/HeJ mouse peritoneal Mphi. Tumor necrosis factor-producing activity was measured by the L929 assay activated with FK-23 (a preparation of heat-killed Enterococcus faecalis). Tumor necrosis factor-producing activity of C3H/HeJ mouse peritoneal Mphi incubated with peritoneal fluid was suppressed in 14 endometriosis patients. Interestingly, in nine endometriosis patients, tumor necrosis factor-producing activity was much lower than seen with mouse peritoneal Mphi incubated with corticosterone. Peritoneal fluid contains suppressive properties for the activation of peritoneal Mphi, which might allow the implantation of free endometrial cells or the metaplastic phenomena stimulated by retrograde menstruation.

R-025. Effect of peritoneal fluid from fertile women and from infertile women with endometriosis or unexplained infertility on the acrosome reaction of capacitated spermatozoa

R-025. Effect of peritoneal fluid from fertile women and from infertile women with endometriosis or unexplained infertility on the acrosome reaction of capacitated spermatozoa

Effect of peritoneal fluid from patients with mild and severe endometriosis on endometrial stromal cell proliferation

Effect of peritoneal fluid from patients with mild and severe endometriosis on endometrial stromal cell proliferation

In vitro culture of endometrial stromal and gland cells as a model for endometriosis: The effect of peritoneal fluid on proliferation

To test the hypothesis that the cell-free fraction of PF from women with endometriosis affects the proliferation of endometrial epithelial and stromal cells in vitro. A cell biologic and immunohistochemical study. University teaching hospital. Premenopausal women undergoing laparoscopy and women with histologically normal endometrium undergoing hysterectomy were selected. Peritoneal fluid (PF) and serum were collected at laparoscopy. Endometrial epithelial and stromal cells were obtained by enzymic dissociation of tissue, and epithelium was separated from stromal cells by sieving. Epithelial and stromal cell populations were purified by removal of contaminating cells using Thy-1-and CD-45-labeled immunomagnetic beads. Isolated endometrial gland and stromal cells were cultured in the presence of PF or serum from women with and without endometriosis. Cell proliferation was assessed by measurement of incorporation of 3[H]thymidine after 48 hours in culture. Isolated endometrial gland and stromal cells were able to proliferate in vitro. The proliferative effect of PF or sera from women with endometriosis did not differ significantly from normal controls. We conclude that PF from women with endometriosis does not have an additional mitogenic effect compared with women without endometriosis. It may be postulated that the endometrium from women with endometriosis responds differently to the effects of PF.