Importance portance of reactive oxygen species in the peritoneal fluid of women with endometriosis or idiopathic infertility

Objective: To determine whether reactive oxygen species in peritoneal fluid might be a factor in infertility.Design: Prospective study.Setting: Andrology laboratory and gynecology clinic at a tertiary care facility.Patient(s): Women with endometriosis (n = 15) or idiopathic infertility (n = 11) who under-went laparoscopy for infertility. Patients undergoing tubal ligation served as controls (n = 13).Intervention(s): Aspiration of peritoneal fluid.Main Outcome Measure(s): Reactive oxygen species levels, presence of polymorphonuclear granulocytes, and leukocyte distribution in peritoneal fluid.Result(s): Reactive oxygen species were present in the peritoneal fluid of patients with endometriosis, idiopathic infertility, and tubal ligation. Levels of reactive oxygen species did not show a statistically significant difference between patients with endometriosis and the control group in either unprocessed or processed (cell-free) peritoneal fluid, but did differ significantly between patients with idiopathic infertility and controls in process peritooneal fluid. Polymorphonuclear granulocytes (> 1 × 10⁸/mL) were not present in the peritoneal fluid of any patient. Macrophage concentrations of peritoneal fluid did not differ significantly between controls and patients with endometriosis or idiopathic infertility.Conclusion(s): Reactive oxygen species in the peritoneal fluid may not affect fertility directly in women with endometriosis; however, they may have a role in patients with idiopathic infertility.     

Increased granulocyte chemotactic protein-2 concentrations in peritoneal fluid of women with endometriosis

BACKGROUND: To evaluate the release of granulocyte chemotactic protein-2 (GCP-2) into peritoneal fluid in women with endometriosis, we measured its concentration with reference to the disease stage and the phase of the menstrual cycle. METHODS: Surgery was scheduled in the proliferative or secretory phase of the menstrual cycle for 64 women with endometriosis (n = 38) or cystadenomas (n = 26). GCP-2 concentrations in the peritoneal fluid were measured using an enzyme-linked immunosorbent assay. RESULT: Our findings indicated elevated concentrations of GCP-2 in peritoneal fluid from women with endometriosis during the proliferative phase, which were positively correlated with the stage of endometriosis. CONCLUSION: Inflammation associated with endometriosis may be involved in the pathogenesis of the disease through increasing levels of peritoneal fluid GCP-2.     

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.     

Increased levels of oxidative stress markers in the peritoneal fluid of women with endometriosis

Objective

To evaluate 8-hydroxy-2-deoxyguanosine (8-OHdG) and 8-isoprostane levels in the peritoneal fluid (PF) of women with endometriosis.

Study design

One hundred and ten women with laparoscopically and histopathologically confirmed endometriosis and, as reference groups, 119 patients with simple serous (n = 78) and dermoid (n = 41) ovarian cysts were studied. Peritoneal fluid 8-OHdG and 8-isoprostane concentrations were evaluated by enzyme-linked immunosorbent assays.

Results

8-OHdG and 8-isoprostane levels in peritoneal fluid were significantly higher in patients with endometriosis compared with the reference groups. Higher PF 8-OHdG and 8-isoprostane concentrations were observed in patients with advanced stages of endometriosis. A statistically significant positive correlation was found between 8-OHdG and 8-isoprostane levels in peritoneal fluid.

Conclusion

Endometriosis induces greater oxidative stress and frequent DNA mutations in peritoneal fluid than nonendometriotic ovarian cysts. The most severe oxidative stress occurs in the peritoneal cavity of women with more advanced stages of the disease.     

Investigation of glutathione concentrations in peritoneal fluid from women with and without endometriosis

Changes in the peritoneal fluid (PF) environment have been implicated in the pathogenesis of endometriosis as well as in the decrease of fertility. OBJECTIVE: To evaluate the concentration of glutathione in PF of women with endometriosis. PATIENTS: Twenty-one patients with endometriosis (I or II rAFS stage, n=11; III or IV rAFS stage, n=10), and 29 patients with follicular or dermoid ovarian cysts (n=17 and n=12, respectively). RESULTS: Mean (+/-S.D.) PF glutathione concentration was 0.22+/-0.01 micromol/ml in patients with minimal or mild endometriosis, 0.21+/-0.05 micromol/ml in women with III or IV stage of the disease, 0.24 +/- 0.03 micromol/ml in women with follicle ovarian cysts, and 0.23+/-0.05 micromol/ml in patients with dermoid tumors of ovaries. No significant difference in the peritoneal glutathione level was found between the groups. CONCLUSION: These results suggest that PF glutathione is not involved in the progression of endometriosis.     

Cytotoxic activity of NK cells in the presence of the peritoneal fluid fro women with unexplained infertility and with endometriosis related infertility

We examined peritoneal fluid obtained during laparoscopy in the aspect of its influence on cytotoxic activity of NK cells. We showed significant suppression of this activity in women with unexplained infertility. The suppression of NK activity was also present in women fertile and infertile both, with severe endometriosis. These results may indicate the immunological disorders in unexplained infertility and in endometriosis-related infertility.     

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.     

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).     

Relation between macrophages in peritoneal fluid of endometriosis and infertility]

Peritoneal fluid was collected from 18 endometriosis-associated infertile patients and 14 unexplained infertile women. The cells were counted in a hemacytometer and subjected to morphologic analysis. The phagocytic activity on Candida Albicans and on Sperms was evaluated and the activity of acid phosphatase in the pelvic fluid and within the macrophages were measured. The results showed that the endometriosis samples have macrophages of large size as compared with the macrophages found in unexplained infertile or fertile women, the concentration of pelvic macrophages in endometriosis is higher, the phagocytic and bactericidal activity is also higher and the macrophages in endometriosis engulf sperms as well. The accentuated activity of pelvic macrophages may be associated with the infertility in 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.     

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.     

促性腺激素释放激素激动剂对子宫内膜异位症不孕患者在位内膜一氧化氮合酶表达的影响

目的探讨一氧化氮合酶(NOS)在子宫内膜异位症(内异症)不孕患者在位子宫内膜(在位内膜)中的表达及促性腺激素释放激素激动剂(GnRHa)对其表达的影响。方法采用免疫组化方法及蛋白印迹方法,对30例Ⅲ～Ⅳ期内异症不孕患者(内异症组)及19例有正常生育史的宫颈原位癌患者(对照组)的在位内膜进行内皮型NOS(eNOS)及诱生型NOS(iNOS)表达的定位及半定量检测,内异症组中18例行GnRHa治疗(治疗组);采用电化学发光法,同时测定患者血清雌二醇及孕酮水平。结果eNOS在子宫内膜的腺上皮、腔上皮及血管内皮细胞中均有表达;iNOS仅在子宫内膜的腺上皮和间质细胞中微弱表达。eNOS在增生期早、中、晚期和分泌期早、中、晚期在位内膜中的相对表达水平,内异症组分别为0.30±0.04,0.40±0.03,0.49±0.03,0.43±0.04,0.55±0.04和0.48±0.03,治疗组分别为0.22±0.03,0.37±0.03,0.45±0.04,0.35±0.05,0.50±0.03和0.41±0.00,对照组分别为0.21±0.03,0.33±0.03,0.45±0.04,0.40±0.03,0.47±0.05和0.41±0.03。在整个月经周期中,内异症组患者在位内膜eNOS相对表达水平持续性高于对照组患者,但仅在增生期早、中期及分泌期中、晚期两组比较,差异有统计学意义(P<0.05);与内异症组比较,治疗组在位内膜eNOS相对表达水平均有所下降,但仅在增生期早期及分泌期早、中期比较,差异有统计学意义(P<0.05)。3组子宫内膜中均未检出iNOS的表达。血清雌二醇或孕酮水平与子宫内膜eNOS表达水平均呈正相关(P<0.01)。结论eNOS在内异症患者在位内膜中的高表达,可能与其发病及其所致的不孕有关,GnRHa治疗可降低内异症患者在位内膜eNOS的表达。     

Etiology of infertility in monkeys with endometriosis: measurement of peritoneal fluid prostaglandins

To study the relationship between peritoneal fluid prostaglandins and infertility associated with endometriosis, we autografted endometrial or adipose tissue to the pelvic peritoneum in 21 monkeys. Peritoneal washings were collected prior to tissue transplantation and during a subsequent laparotomy performed for biopsy of the implants. Monkeys were mated and peritoneal washings were collected during three subsequent cycles. The content of prostaglandin F2 alpha (PGF2 alpha) in adipose tissue autografts was significantly less (p less than 0.05) than in endometrial tissue autografts. The PGF2 alpha concentration in peritoneal fluid increased significantly (p less than 0.05) only in monkeys that developed moderate or severe endometriosis. Prostaglandin E levels in tissue autografts or peritoneal fluid were similar in all animals. Infertility in monkeys with endometriosis was associated with luteinized unruptured follicles, luteal phase defects, and pelvic adhesions. Although PGF2 alpha concentrations in peritoneal washings obtained during these cycles were increased in comparison with those of ovulatory cycles, the difference was not significant. A relationship between spontaneous abortion and prostaglandin concentrations in peritoneal fluid was not established.     

Inducible nitric oxide synthase expression by peritoneal macrophages in endometriosis-associated infertility.

OBJECTIVE: Determine whether peritoneal macrophages from women with endometriosis-associated infertility express more inducible nitric oxide synthase (NOS2) and produce more NO than fertile controls. DESIGN: Unblinded clinical study. PATIENT(S): Nine infertile women with endometriosis and nine normal fertile women undergoing laparoscopy.Intervention(s): Peritoneal fluid and macrophages were collected. Cells were also cultured with the NOS2 inducers interferon-alpha (IFN-alpha) or IFN-gamma plus lipopolysaccharide (LPS). MAIN OUTCOME MEASURE(S): Peritoneal fluid NO levels, peritoneal macrophage NOS activity, and peritoneal macrophage NOS2 protein expression. RESULT(S): NOS enzyme activity was higher in peritoneal macrophages from endometriosis patients. Immunoblots demonstrated NOS2 protein only in peritoneal macrophages from women with endometriosis. Peritoneal fluid NO concentration was similar in the two groups, but total peritoneal fluid NO content was higher in endometriosis patients. After 3 days' culture, peritoneal macrophages from women with endometriosis produced more NO in response to IFN-alpha or IFN-gamma plus LPS than controls. CONCLUSION(S): Peritoneal macrophages from women with endometriosis-associated infertility express higher levels of NOS2, have higher NOS enzyme activity, and produce more NO in response to immune stimulation in vitro. As high levels of NO adversely affect sperm, embryos, implantation, and oviductal function, reducing peritoneal fluid NO production or blocking NO effects may improve fertility in women with 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.     

Prostaglandins in peritoneal fluid of women with minimal and mild endometriosis]

Peritoneal fluid from 52 women with minimal and mild endometriosis was aspirated at laparoscopy and PGs were analyzed. Peritoneal lesions were classified into black, red and white lesions and peritoneal pockets, and were excised and pathological examination performed. Patients were classified into the black lesion group (n = 17) and the red lesion group (n = 35) according to the main colour of the lesions. 1. Endometrial gland and stroma were found in 78% of black lesions, 53% of red lesions, 60% of white lesions and 62% of peritoneal pockets. Infiltration of lymphocytes was found in 74% of black lesions, 83% of red lesions, 80% of white lesions and 77% of peritoneal pockets. 2. The fluid volume of the red lesions group was significantly higher than that of the control group (p less than 0.01). But the fluid volume of the black lesions group was insignificant as compared with that of the control group. 3. The PGE2 and PGF2 alpha concentrations in fluid from the red lesion group were significantly higher than not only the control group (p less than 0.001, p less than 0.05) but also the black lesion group (p less than 0.01, p less than 0.05). Only the PGE2 concentration in fluid from the black lesion group was significantly higher than that of the control group (p less than 0.001). 4. In the red lesion group which had no infertility factors without endometriosis, 20 of 25 patients achieved pregnancy (80%) in a one-year follow up. This was significantly higher than in the black lesions group (44%).(ABSTRACT TRUNCATED AT 250 WORDS)     

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).     

Plasma and peritoneal fluid levels of CA 125 in women with endometriosis

Determined by a sandwich, solid-phase radioimmunoassay with mouse monoclonal antibody, OC 125, plasma CA 125 levels were significantly elevated in stage III (mean +/- SEM = 32.7 +/- 5.2 U/ml, n = 17, p = less than 0.01) and stage IV endometriosis (37.2 +/- 10.5 U/ml, n = 6, p = less than 0.005) compared with levels during the follicular (15.9 +/- 1.5 U/ml, n = 12) and secretory phases (15.8 +/- 1.3 U/ml, n = 15) of control women and users of oral contraceptives (15.5 +/- 1.2 U/ml n = 10). However, CA 125 levels were not significantly elevated in women with stage I (16.6 +/- 2.0 U/ml, n = 28) or stage II endometriosis (17.9 +/- 2.1 U/ml, n = 13). Peritoneal fluid levels of CA 125 (n = 14) were significantly higher (2 to 9.3 times) than the corresponding paired plasma levels in participants with stage I, II, or III endometriosis. In patients treated with danazol (n = 10) or buserelin (n = 17), plasma CA 125 levels decreased significantly by midtherapy, remained suppressed at the end of therapy, but rebounded to near pretreatment levels 6 months after treatment was completed. With gestrinone therapy a similar decline was observed but became significant only at the end of therapy. Our findings indicate that elevated plasma CA 125 levels may prove useful in the management of endometriosis.