Lipid peroxidation in the peritoneal fluid of infertile women with peritoneal endometriosis

OBJECTIVE: To assess the level of lipid peroxidation in the peritoneal fluid of infertile women with peritoneal endometriosis and of fertile disease-free controls. STUDY DESIGN: Level of lipid peroxidation (malondialdeyde, malondialdeyde with copper addition, and cholest-3,5-dien-7-one) was measured in the peritoneal fluid obtained from 21 women with endometriosis-related infertility and from 21 fertile women having tubal ligation. RESULTS:: The level of lipid peroxidation did not differ significantly (P > 0.05) according to the stage of endometriosis. The level of lipid peroxidation (malondialdeyde, malondialdeyde with the addition of copper, and cholest-3,5-dien-7-one) did not differ significantly (P > 0.05) between patients with endometriosis-related infertility (0.07 nmol/ml, 0.34 nmol/ml, 0.24 microg/ml, respectively) and disease-free controls (0.04 nmol/ml, 0.21 nmol/ml, 0.25 microg/ml, respectively). CONCLUSION: The level of lipid peroxidation did not differ between women with endometriosis-related infertility and fertile disease-free controls, suggesting that increased reactive oxygen species may not be one of the factors responsible for compromised fertility in patients with endometriosis.     

Prostanoids in peritoneal fluid of infertile women with pelvic endometriosis and PID

Peritoneal fluid collected at celioscopy in infertile subjects was assayed for steroids and several prostanoids (PGE2, PGF2, TXB2, LTB4) as part of a study into pathophysiology of the female reproductive tract. Prostaglandins, produced massively in the pelvis, might interfere with fertility through various mechanisms (alterations in the egg implantation, follicle genesis, luteinization as well as tubal disorders). Our study of 54 patients showed a marked increase only of TXB2 out of the prostanoids assayed in overall endometriosis. In pelvic flogosis peritoneal LTB4 (and TXB2) were considerably increased if related to controls. This would suggest their role in the ethiopathogenesis of unexplained infertility (in relation to these pathologic patterns).     

Elevated peritoneal fluid luteinizing hormone and prolactin concentrations in infertile women with endometriosis

In this study, we compared (Mann-Whitney U-test) the peritoneal fluid FSH, LH and PRL levels, measured by RIA, at the follicular and luteal phases of the menstrual cycle in women with (n = 43; age 25-44 years) and with no evidence of endometriosis (n = 35; age 25-39 years) who were considered as controls. Both follicular and luteal phase FSH concentrations of women with endometriosis were not statistically different (n = 22 vs 18; 0.32-5.8 vs 0.50-8.2 IU/l, P = 0.247; n = 13 vs 14; 0.6-6.5 vs 0.66-6.7 IU/l, P = 0.604) compared to their respective controls. In contrast to FSH, the concentrations of LH at follicular (n = 19 vs 17; 3.1-34.2 vs 2.3-12.2 IU/l, P = 0.01) and luteal (n = 17 vs 15; 2.1-95.4 vs 1.3-17.9 IU/l, P = 0.02) phases of the test group was significantly elevated at both phases of the cycle. With respect to differences in PRL concentrations at follicular phase no significant change (n = 21 vs 16; 1030-5800 vs 1305-4650 mIU/l; P = 0.255) was observed. The greatest difference in luteal PRL concentrations (P = 0.007) was obtained between the women with endometriosis and controls (n = 17 vs 17; 1895-8600 vs 1041-5000 mIU/l). The results suggest that disordered synchronization of neuroendocrine mechanisms controlling LH and PRL may be the underlying abnormality causing infertility in our group of patients with endometriosis.     

Cytokine profiles in serum and peritoneal fluid from infertile women with and without endometriosis

OBJECTIVE: To study the serum and peritoneal fluid cytokine profiles in infertile women with minimal/mild active endometriosis. METHODS: Fifty-seven consecutive infertile women undergoing laparoscopy for unexplained infertility had peritoneal fluid and serum samples obtained at the time of laparoscopy. The levels of interleukin-6 (IL-6), interleukin-8 (IL-8), interleukin-1 beta (IL-1 beta), vascular endothelial growth factor (VEGF), tumor necrosis factor-alpha (TNF-alpha), monocyte chemotatic protein-1 (MCP-1), RANTES, platelet derived growth factor (PDGF), soluble Fas (sFas), and soluble Fas Ligand (sFasL) in peritoneal fluid and serum were measured to compare the concentration in both biological fluids, in women who have minimal/mild red endometriosis using women with no endometriosis as controls. RESULTS: Peritoneal fluid levels of MCP-1, IL-8 and IL-6 were significantly higher in the endometriosis group (P < 0.012, P = 0.003, and P = 0.015, respectively). There was no significant difference in the peritoneal fluid levels of IL-1 beta, TNF-alpha, RANTES, VEGF, PDGF, sFas and sFasL in the two groups. Although serum levels of IL-8 were higher in women with endometriosis, the difference was not significant (P = 0.07). Serum levels of PDGF, IL-6, RANTES, IL-1 beta, TNF-alpha, and sFas, were not significantly different in the two groups. CONCLUSION: The elevated levels of MCP-1, IL-6, and IL-8 in peritoneal fluid but not serum may indicate the importance of local macrophage activating factors in the pathogenesis of endometriosis.     

Pyruvate reduces in vitro the embryotoxic effect of peritoneal fluid from infertile women with endometriosis

OBJECTIVES: To ascertain the embryotoxicity of peritoneal fluid from infertile women with endometriosis (PF-E), on mouse embryos in culture and to examine the effect of pyruvate in the culture medium on PF-E induced embryotoxicity. STUDY DESIGN: Blood-free peritoneal fluid samples were obtained during laparoscopic investigation for infertility from 21 infertile women with endometriosis. The severity of endometriosis ranged from minimal or mild (PF-min to mild-E; n=7), moderate (PF-mod-E; n=7), to severe (PF-sev-E; n=7). Peritoneal fluid samples were centrifuged at 600 x g for 10 min and 4 degrees C, and the supernatant was incubated at 56 degrees C for 30 min in a water bath to inactivate the complement protein. Mice were super ovulated with intraperitoneal injection (IP) of 5IU of pregnant mare serum gonadotrophin and human chorion gonadotrophin. Twenty-four hours after confirmation of mating two-cell mouse embryos were obtained. They were then cultured in modified Whitten's medium (mWM) with peritoneal fluid from patients with endometriosis, and either in the absence or presence of excess pyruvate (0.062 mmol(-1)). Embryos were cultured for 72 h. RESULTS AND CONCLUSION: Addition of 5% PF-E significantly (p<0.001) suppressed embryo growth at 24, 48, and 72 h of culture and the degree of suppression correlated with the severity of the disease. The presence of 0.062 mmol(-1) pyruvate in the culture medium significantly (p<0.001) reduced the embryotoxicity of PF-min to mild-E and PF-mod-E at each stage of development, but was only seen at 24h of culture (p<0.001) in cultures with PF-sev-E even when the concentration of pyruvate in the medium was increased to 0.31 mmol(-1). This study confirms the embryotoxicity of PF-E in vitro, which was reduced by the presence pyruvate in the culture medium, particularly in cultures containing fluid from women with endometriosis of minimum or mild to moderate severity.     

Lymphocyte activity in the presence of peritoneal fluid from fertile women and infertile women with and without endometriosis

Peritoneal fluid from women with endometriosis, unexplained infertility, and fertile controls were compared to one another and to normal human serum for effects on lymphocyte proliferation in vitro. Peritoneal fluid samples were also assayed for both interleukin-1 and interleukin-2. All peritoneal fluid samples significantly enhanced lymphocyte proliferation in both mitogen-stimulated and unstimulated cultures compared with serum controls. Mitogen-induced leukocyte proliferation was higher in the presence of peritoneal fluid from women with endometriosis compared with other samples. Five out of 23 samples from endometriosis patients contained elevated levels of interleukin-1 and three out of 23 contained elevated levels of interleukin-2. Six out of eight peritoneal fluid samples from unexplained infertility patients also had elevated levels of interleukin-2; samples from fertile women did not contain elevated levels of either cytokine. Our data indicate that peritoneal fluid from women with endometriosis and unexplained infertility support the activation and proliferation of lymphocytes. Leukocyte products may locally affect the progression of disease and fertility.     

Histologic study of peritoneal endometriosis in infertile women

The present study included 118 patients undergoing a laparoscopy for infertility. In 86 patients with laparoscopically diagnosed endometriosis (group I), biopsies were taken from areas of apparent endometriosis (n = 86) and from a visually normal peritoneum (n = 52). Histology reveals the presence of endometriosis in 93% of positive sites and in 13% of negative sites. In 32 patients without endometriosis at laparoscopy (group II), biopsies were taken from normal uterosacral ligaments (n = 32). Endometriosis was observed in 6% of cases. Despite the increased ability to detect pigmented and nonpigmented endometriotic lesion, histological study revealed the presence of endometriosis in normal peritoneum in 13% (group I) and 6% (group II) of cases.     

Cellular components in peritoneal fluid in infertile patients with and without endometriosis

Cellular components in peritoneal fluid of infertile patients with and without endometriosis were evaluated in 102 patients with Wright's-Giemsa and Papanicolaou stains. The secretory activity of these cells was studied indirectly by assaying acid phosphatase, prostaglandin (PG) F2 alpha and PGE2 and complement components C3c and C4. The results showed that macrophages and lymphocytes were the dominant cells in peritoneal fluid of these patients. These cells were significantly increased in endometriosis patients, as compared with control subjects. In addition, peritoneal fluid acid phosphatase, PGF2 alpha and PGE2, and complement components C3c and C4 were significantly increased in patients with endometriosis. These cellular changes and their activation in peritoneal fluid may explain infertility associated with endometriosis.     

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.     

Fractalkine in the peritoneal fluid of women with endometriosis.

OBJECTIVE: The objective of this study was to evaluate the presence of fractalkine in the ascites and the association between fractalkine levels in the ascites and endometriosis. METHODS: Peritoneal fluids and peripheral blood samples were obtained from patients undergoing laparoscopy for infertility work-up or laparoscopic cystectomy. Three samples of peritoneum were obtained from patients undergoing hysterectomy. Western blotting, RT-PCR and immunohistochemistry were performed. RESULTS: Fractalkine protein was detected in the ascites. Positive staining was confirmed in peritoneal surface cells and perivascular cells of the peritoneum. CX3CR1 positive cells were present in the cells in the peritoneal fluid. The fractalkine concentrations in the ascites of patients with endometriosis were lower than those without endometriosis. There was no significant difference between serum fractalkine levels in patients with and without endometriosis. CONCLUSION: The decreased level of fractalkine found in the peritoneal fluid of patients with endometriosis may contribute to the pathogenesis of endometriosis.     

Comparison of cytokine levels and embryo toxicity in peritoneal fluid in infertile women with untreated or treated endometriosis.

OBJECTIVE: Our aim was to examine the relationship between the levels of cytokines in peritoneal fluid and its embryo toxicity. STUDY DESIGN: The levels of interleukin-1 and tumor necrosis factor were measured in peritoneal fluid from infertile women who did not have endometriosis (n = 21), who had untreated endometriosis (n = 19), and who had undergone medical treatment for endometriosis (n = 10). Embryo toxicity was investigated in mouse two-cell embryos cocultured with the oviducts in culture media that contained various concentrations of peritoneal fluid. RESULTS: The levels of cytokines were significantly higher in the peritoneal fluid from women who had untreated endometriosis than in women who did not have endometriosis, but they were extremely low in women who had undergone medical treatment with either danazol or buserelin. The peritoneal fluid from women who had untreated endometriosis adversely affected the cleavage of mouse two-cell embryos. After medical treatment the embryo toxicity of the peritoneal fluid was almost undetectable. CONCLUSION: These results offer some theoretic bases in support of medical treatment to improve reproductive performance in infertile women who have endometriosis.     

Glutathione peroxidase concentration in the peritoneal fluid from infertile women

OBJECTIVES: To assess the concentration of Plasma Glutathione Peroxidase (plGPx) in the peritoneal fluid (PF) of patients with unexplained infertility and infertile women with minimal and mild endometriosis. MATERIALS AND METHODS: 33 women were studied, including 8 infertile women with minimal or mild endometriosis, 15 patients with unexplained infertility and 10 patients with tubal occlusion (a reference group). Concentration of plGPx was measured in the PF using a commercially available ELISA kit (Oxis Inc.). RESULTS: The plGPx concentration was significantly (p = 0.04) lower in PF from women with unexplained infertility (846 +/- 177 ng/ml) compared to the reference group (1023 +/- 238 ng/ml), but did not differ significantly (p = 0.25) between women with endometriosis (918 +/- 81 ng/ml) and patients with tubal infertility. CONCLUSIONS: Our results suggest that low peritoneal plGPx concentration may play a role in the pathogenesis of infertility.     

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.     

Total antioxidant status of peritoneal fluid in infertile women

OBJECTIVE: To determine whether impairment of the antioxidant systems of peritoneal fluid might be a factor responsible for infertility. STUDY DESIGN: Total antioxidant status was measured in peritoneal fluid obtained from 18 infertile women suffering from minimal or mild endometriosis, 23 patients with unexplained infertility, 12 women with tubal infertility and 13 fertile women. RESULTS: Total antioxidant status was significantly lower in peritoneal fluid from women with unexplained infertility (0.49+/-0.21 mmol/l) compared to both fertile patients (0.67+/-0.24 mmol/l, P=0.02) and women with tubal infertility (0.76+/-0.26 mmol/l, P=0.001). Peritoneal fluid total antioxidant status did not differ significantly between patients with endometriosis (0.61+/-0.2 mmol/l), tubal infertility and the fertile group (P>0.05). CONCLUSIONS: Our results suggest that low antioxidant status in peritoneal fluid may play a role in the pathogenesis of infertility.     

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.     

Characterization of leukocyte subpopulations in the peritoneal fluid of women with endometriosis

Monoclonal antibodies identifying leukocytes subpopulations were applied to smears of laparoscopically collected peritoneal fluid leukocytes and parallel samples of peripheral blood leukocytes from women with endometriosis (n = 33), those with unexplained infertility (n = 9), and fertile controls (n = 8). Peripheral blood leukocyte profiles in all groups were indistinguishable from reported normal values. Peritoneal fluid leukocyte profiles were observed to be different between groups. The most significant elevations in total leukocytes, macrophages, helper T lymphocytes and natural-killer cells were observed in women with stage I and II endometriosis. Significantly elevated levels of total leukocytes, macrophages, and T lymphocytes were also observed in peritoneal fluid from women with unexplained infertility. The results from this study indicate that the peritoneal environment is immunologically dynamic and suggest that cellular immune mechanisms may contribute to reproductive failure in women with endometriosis and unexplained infertility.     

Serum and peritoneal fluid proteins in women with and without endometriosis

We examined the proteins in serum and peritoneal fluid of women with endometriosis (and of healthy controls) for evidence of an autoimmune response that might account for their impaired fertility. No antibodies against endometrial glycoproteins or against "progestin dependent endometrial protein" (PEP) were found in any serum or peritoneal fluid sample. Levels of PEP were not different in serum from women with moderate to severe endometriosis (n = 6), with mild endometriosis (n = 21), or from disease-free cycling controls (n = 19). PEP levels in peritoneal fluid from mild endometriosis and from controls did not differ but were elevated ten times in fluid obtained in the secretory phase from women with moderate to severe disease. This suggests that PEP levels in peritoneal fluid reflect the extent of ectopic endometrial growth. The salient finding was a heretofore undescribed protein (mol wt 70,000) in secretory phase peritoneal fluid samples (18/20) and its absence during the proliferative phase (0/35).     

Decreased lactoferrin levels in peritoneal fluid of women with minimal endometriosis

OBJECTIVE: The aim of the study was to evaluate for the presence of lactoferrin (LTF) in peritoneal fluid (PF) of women with and without endometriosis. PATIENTS: Seventy-eight women were studied, including 49 women with endometriosis and, as a reference group, 29 patients with functional follicle ovarian cysts. RESULTS: Lactoferrin levels were detectable in all peritoneal fluid samples. Women with minimal endometriosis had lower PF lactoferrin concentrations compared to both patients with high revised American Fertility Society classification scores and women with follicle ovarian cysts. No significant difference in the peritoneal LTF levels was found between patients with stage II endometriosis, stage III or IV endometriotic disease and women with functional cysts of ovaries. CONCLUSIONS: Owing to its antibacterial properties lactoferrin is probably an important defense factor in the peritoneal cavity, however its role in the pathogenesis of endometriosis remains enigmatic.