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.     

超长方案降调节后LH添加时机对子宫内膜异位症患者IVF-ET的影响

目的:探讨在子宫内膜异位症(EMs)合并不孕患者体外受精-胚胎移植(IVF-ET)超促排卵过程中,经超长方案长效GnRH-a降调节后添加LH的合适时机。方法:回顾分析2010年1月至2012年4月在我中心行IVF-ET助孕的118例不孕合并EMs患者,共123个周期。所有周期均采用超长方案,根据添加LH时卵泡的直径分为3组:10～14mm(A组),14～16mm(B组)及≥16mm(C组),比较3组患者的卵巢反应性及IVF妊娠结局。结果:A组的P水平(0.72±0.63)显著低于B组(1.08±0.72)和C组(1.11±0.71)。A组的优质胚胎率和妊娠率分别为75.81%和55.00%,显著高于B、C组;A组获卵数(7.42±4.86)显著低于C组(11.34±5.71)(P<0.05)。结论:EMs合并不孕患者经超长方案长效GnRH-a降调节后,尽早添加外源性LH,可有效降低孕酮水平,提高优质胚胎率和妊娠率。     

Impaired insulin action on granulosa-lutein cells in women with polycystic ovary syndrome and insulin resistance.

We studied the in vitro response to insulin of granulosa-lutein cells derived from patients with polycystic ovary syndrome (PCOS) and clinically defined insulin resistance. Insulin sensitivity was assessed by continuous infusion of glucose with model assessment test (CIGMA). Insulin resistant (PCOS-IR; n = 8), noninsulin resistant (PCOS-NIR; n = 9) patients with PCOS, and women with tubal factor infertility (TF; n = 8) underwent controlled ovarian stimulation with long-term gonadotropin-releasing hormone (GnRH) agonist, recombinant follicle stimulating hormone (FSH), and in vitro fertilization. Primary cultures of granulosa-lutein cells were incubated with insulin (10, 100, 500 ng/ml) and/or luteinizing hormone (LH) (10, 100 ng/ml) in the presence of low density lipoprotein (100 micrograms/ml). The progesterone and lactate accumulation were measured in the culture medium. LH potently stimulated the progesterone secretion in all groups. Insulin alone had no effect on progesterone release in any of the groups, but stimulated lactate formation in the PCOS-NIR and TF groups. Insulin augmented the effect of LH on progesterone secretion selectively in the PCOS-NIR group. The expression of the insulin receptor was determined by Western blotting in separate cultures of granulosa-lutein cells, and showed receptor down-regulation in the PCOS-IR patients. We infer that the in vitro effect of insulin on progesterone and lactate release by granulosa-lutein cells is impaired in insulin resistant PCOS patients.     

Impaired insulin action on granulosa-lutein cells in women with polycystic ovary syndrome and insulin resistance

We studied the in vitro response to insulin of granulosalutein cells derived from patients with polycystic ovary syndrome (PCOS) and clinically defined insulin resistance. Insulin sensitivity was assessed by continuous infusion of glucose with model assessment test (CIGMA). Insulin resistant (PCOS-IR; n = 8), non-insulin resistant (PCOS-NIR; n = 9) patients with PCOS, and women with tubal factor infertility (TF; n = 8) underwent controlled ovarian stimulation with long-term gonadotropin-releasing hormone (GnRH) agonist, recombinant follicle stimulating hormone (FSH), and in vitro fertilization. Primary cultures of granulosa-lutein cells were incubated with insulin (10, 100, 500 ng/ml) and/or luteinzing hormone (LH) (10, 100 ng/ml) in the presence of low density lipoprotein (100 μg/ml). The progesterone and lactate accumulation were measured in the culture medium. LH potently stimulated the progesterone secretion in all groups. Insulin alone had no effect on progesterone release in any of the groups, but stimulated lactate formation in the PCOS-NIR and TF groups. Insulin augmented the effect of LH on progesterone secretion selectively in the PCOS-NIR group. The expression of the insulin receptor was determined by Western blotting in separate cultures of granulosa-lutein cells, and showed receptor down-regulation in the PCOS-IR patients. We infer that the in vitro effect of insulin on progesterone and lactate release by granulosa-lutein cells is impaired in insulin resistant PCOS patients.     

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.     

The assessment of dendritic cells cultured from peritoneal fluid macrophages: first report and new perspectives in the treatment of endometriosis

Dendritic cells represent discrete leukocyte subpopulation of specialist or "professional" antigen-presenting cells (APC). They play a crucial role in the activation of naive T cells "in vivo" They have monocyte/macrophages origin. There are no data in literature on the presence of dendritic cells derived from peritoneal fluid monocytes/macrophages. In our study we tried to culture PF macrophages from patients who undergone surgery so that to obtain dendritic cells. PF was aspirated during laparoscopy from patients with endometriosis, unexplained infertility or benign noninflammatory ovarian tumor. Peritoneal macrophages were isolated using adherence method then were cultured and stimulated with GM-CSF and IL-4. Phenotype of cultured cell was estimated using flow cytometry after incubation with monoclonal antibodies CD45/14, CD 40/HLA-DR, CD28/3, CD3/40L, CD25/5 and CD69/HLA-DR. Morphology of cultured cells was confirmed microscopically after May-Grunvald-Giemsa staining. PF leukocytes concentration varied from 1.2 x 10(6) cells/mm3 to 22.6 x 10(6) cells/mm3. Cultured monocytes/macrophages from PF had morphology typical for dendritic cells. We also found that only dendritic cells from patients with endometriosis had higher expression HLA-DR antigen (93.6% of cells) and low expression of CD40 (2.7% of cells) on their surface in comparison to reference group. It is worthy to notify that dendritic cells from patients with endometriosis expressed also CD25 antigen characteristic for T leukocytes. To our data it is the first report in literature on dendritic cells obtained from PF macrophages.     

Effects of previous ovarian surgery for endometriosis on the outcome of assisted reproduction treatment

Endometriosis affects 2-50% of women at reproductive age. Surgery is an option for treatment, but there is no convincing evidence that it promotes a significant improvement in fertility. Also, the removal of ovarian endometrioma might lead to a reduction in the follicular reserve and response to stimulation. Therefore, the aim of this study was to evaluate the effect of previous ovarian surgery for endometriosis on the ovarian response in assisted reproduction treatment cycles and its pregnancy outcome. A total of 61 women, with primary infertility and previously having undergone ovarian surgery for endometriosis, who had received 74 IVF/intracytoplasmic sperm injection (ICSI) cycles, were studied (study group). A further 74 patients with primary infertility who underwent 77 IVF/ICSI cycles within#10; the same period of time, at the same clinic and without previous ovarian surgery or endometriosis were studied as a control group. Patients were matched for age and treatment performed. Patients 35 years with previous ovarian surgery needed more ampoules for ovulation induction (P = 0.017) and had fewer follicles and oocytes than women in the control group (P = 0.001). Duration of folliculogenesis was similar in both groups, as was fertilization rate. A total of 10 patients achieved pregnancy in the study group (34.5%) and 14 (48.3%) in the control group. Although a lower pregnancy rate was observed in patients who had undergone previous ovarian surgery, this difference was not statistically significant (P = 0.424). In conclusion, ovarian surgery for the treatment of endometriosis reduces the ovarian outcome in IVF/ICSI cycles in women >35 years old, and might also decrease pregnancy rates. Therefore, for infertile patients, non-surgical treatment might be a better option to avoid reduction of the ovarian response.     

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.     

Vascular endothelial growth factor in the plasma, follicular fluid and granulosa cells of women with endometriosis submitted to in vitro fertilization--a pilot study.

Vascular endothelial growth factor (VEGF), a potent angiogenic factor that is altered in endometriosis, supports the immunological mechanism involved in this disease. The aim of the present study was to assess VEGF concentration in the plasma, follicular fluid (FF) and culture medium (CM) of granulosa cells from patients with endometriosis submitted to in vitro fertilization (IVF). A case-control study was conducted on 14 patients with endometriosis and 14 women without endometriosis submitted to IVF. Peripheral blood samples were collected before and after administration of human chorionic gonadotropin (hCG), in addition to FF and CM samples. Plasma VEGF levels increased after hCG administration in women with endometriosis and in controls, but were significant only in controls. VEGF levels were lower in FF but were significantly increased in the CM of patients with endometriosis. There was no correlation between VEGF and age, response to ovarian stimulation, oocyte or embryo quality, and pregnancy result. The increase of VEGF levels after hCG in both groups demonstrated a positive effect of this hormone on VEGF. VEGF in the FF and CM presented opposite results in endometriosis, suggesting that granulosa cells may show a different behavior in vivo and in vitro.     

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   

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 alphavbeta3 integrin was measured in the pregnant mouse uterus by immunohistochemistry with in situ hybridization. Leukemia inhibitory factor and the beta3 subunit of alphavbeta3 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 alphavbeta3 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 alphavbeta3 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.     

Effect of ovarian involvement on the frequency of luteinized unruptured follicle in endometriosis.

OBJECTIVE: When we review the current literature on endometriosis and luteinized unruptured follicle (LUF), we see that most of the studies deal with only the association between LUF frequency and the severity of endometriosis. Our purpose was to evaluate the effect of ovarian involvement on LUF frequency in endometriosis and assess the relationship between endometriosis and LUF in infertile women. STUDY DESIGN: This study is a prospective analysis covering a total of 126 infertile women between 22 and 35 years of age who underwent diagnostic laparoscopy from September 1995 to August 1997 in the Department of Obstetrics and Gynecology at Süleyman Demirel University, Isparta, Turkey. Endometriosis was diagnosed in 58 of these patients. All had received a revised American Fertility Society staging score at the time of the laparoscopic diagnosis. Diagnosis of LUF was made when the following criteria were fulfilled: absence of ultrasonic signs of ovulation, and absence of an ovulation ostium on the follicle by laparoscopy, despite increased serum progesterone. Statistical evaluation was performed using chi2 test and Fisher's exact test where appropriate. RESULTS: The prevalence of LUF in mild, moderate and severe endometriosis cases was 13.3, 41.2 and 72.7%, respectively. The LUF frequencies were 45.9% in 37 endometriosis patients with ovarian involvement, 9.5% in 21 cases without ovarian involvement, and 5.9% in 68 cases without endometriosis. A statistically significant difference was observed between the LUF rate in the group with ovarian involvement and that without involvement (chi(2) = 8.06, p < 0.001). CONCLUSION: In summary, in this study we noted a significant increase in LUF frequency in endometriosis patients with ovarian involvement.     

Mild endometriosis and luteal function

The luteal function of 27 patients with mild endometriosis and infertility was compared with that of 50 infertile patients without endometriosis. The incidence of endometrial luteal phase deficiency was similar in both groups of patients. Luteal phase length and plasma levels of progesterone (P), estradiol (E2) and prolactin (PRL) in infertile patients (both groups) were similar to those in a group of 10 fertile women. We conclude that luteal phase defects should not be considered as a primary cause of infertility in mild endometriosis.     

Autoimmune reaction factors of infertility in patients with endometriosis]

The presence of immunoprotein depositions in endometrium and antiendometrial autoantibodies in serum was detected in 59 infertile patients with endometriosis, 45 infertile patients without endometriosis and 11 fertile women by the methods of immunohistochemistry determination and immunodiffusion assay. The immunoprotein depositions in endometrium of endometriosis were significantly increased. The antiendometrial auto-antibodies in serum were positive in 32.2% for infertile patients with endometriosis, 6.7% for infertile patients without endometriosis and 0% for fertile women. This suggested that an autoimmune response might be an etiologic factor of infertility in patients with endometriosis.     

Lipid peroxides, tumor necrosis factor alpha (TNF-alpha) and interferon gamma (IFN-gamma) in peritoneal fluid from infertile women with minimal and mild endometriosis

OBJECTIVES: To assess the concentrations of tumor necrosis factor alpha (TNF-alpha), interferon gamma (IFN-gamma) and lipid peroxides (the marker of free radicals activity) in peritoneal fluid (PF) of infertile women with minimal and mild endometriosis. MATERIALS AND METHODS: 19 women were studied, including 9 infertile women with minimal or mild endometriosis and 10 patients with tubal occlusion (the reference group). Lipid peroxides (malonyldialdehyde and 4-hydroxynonenal), TNF-alpha and IFN-gamma concentrations were measured in the PF using commercially available kits. RESULTS: Concentration of IFN-gamma was detectable in PF of 7 (77.8%) women with endometriosis and in PF from 3 (30%) patients with tubal occlusion. Neither TNF-alpha or lipid peroxides PF concentration differed significantly (p < 0.05) between the groups. In the group with endometriosis we have found a positive correlation (R = 0.77, p = 0.04) between the concentrations of TNF-alpha and IFN-gamma. CONCLUSIONS: Our results suggest that oxidative stress in the PF doesn't appear to play a role in endometriosis-associated infertility.     

Relationship between luteal function and prolactin in infertile women with endometriosis

Luteal function in 44 infertile women with endometriosis were studied with reference to prolactin (PRL) and compared with 34 unexplained infertile women without endometriosis. To assess luteal function, serum progesterone (P4) levels were measured on the 3rd, 7th and 10th days of the luteal phase. On the 7th day, serum estradiol (E2) levels and PRL levels were also determined. The response of PRL secretion to TRH was examined at 30 and 60 after following TRH (500 micrograms, im.) administration. The incidence of hyperprolactinemia (basal PRL level greater than or equal to 25 ng/ml) and latent hyperprolactinemia (peak PRL level in TRH challenge test greater than or equal to 150 ng/ml) were 19% and 31%, respectively, in the endometriosis group and 14% and 33%, respectively in the control group. At the midluteal stage, serum P4 levels in endometriosis group were decreased significantly (p less than 0.05), whereas no difference was found between the serum E2 levels in the endometriosis group and the control. In the endometriosis group, there was no correlation between P4 and E2 levels and abnormal secretion of PRL such as hyperprolactinemia and latent hyperprolactinemia. These results indicate the close association of endometriosis with an inadequate luteal phase. However, it seems that the aberrant secretion of PRL has no relation to the impared luteal function in endometriosis.     

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

The aim of this study was to evaluate and compare the mitogenic effect of peritoneal fluid (PF) from women with mild and severe endometriosis on the endometrial stromal cell proliferation. Increasing concentrations of PF from women with and without mild or severe endometriosis were added to primary endometrial stromal cell cultures and³H-thymidine incorporation was used to assess DNA synthesis in these cultures. PF from women with mild endometriosis induced a statistically significant dose-dependent increase in stromal cell thymidine uptake ranged from 5.8 to 14.5 fold, whereas PF from women with severe endometriosis produced an average 51% inhibition of stromal cell proliferation of compared with cells exposed to non-endometriosis PF or exposed to nutrient medium supplemented with 2.5% calf serum alone. PF samples from patients with stage I endometriosis induced a statistically dose-dependent increase in stromal cell proliferation, whereas PF from patients with stage IV endometriosis caused a significant inhibition.     

Application of IVF in women with endometriosis

Application of IVF at women with endometriosis after pharmacological and surgical therapy hold out hopes to possession of child. Endometriosis occurs three times more often in the patients with primary sterility than in patients with secondary sterility. The presence of a small endometriosis does not reduce the success of the IVF therapy. The women with endometriosis who undergo IVF treatment have similar prospect in compensate with women with tubal sterility. IVF improve fertility in women with mild or minimal endometriosis. But at women after IVF the presence of endometriomas is associated with increased rates of early pregnancy losses. Women with endometriosis marking often antinuclear antibodies and antiphospholipid antibodies than women without endometriosis. To evaluate the 15% of infertile women have endometriosis.     

The stimulus responsible for the peritoneal fluid inflammation observed in infertile women with endometriosis

OBJECTIVE: We tested the hypothesis that menstrual debris from ectopic endometrium is the stimulus responsible for eliciting the peritoneal fluid (PF) inflammation observed in infertile women with endometriosis. DESIGN, SETTING, PATIENTS: The extent of endometriosis was correlated with the PF volume and total PF cell count retrospectively in 135 infertile women with endometriosis. RESULTS: The volume and total cell count were positively correlated, whereas the total cell count was negatively correlated with the extent of endometriosis. Despite a similar negative trend, no statistically significant correlation was noted between the volume and the extent of endometriosis. These relationships did not change when the data were reanalyzed deleting those pathological features contributing to the endometriosis score but not capable of producing intraperitoneal menstrual debris, i.e., adhesions and encapsulated ovarian endometriomas. CONCLUSIONS: These findings indicate that menstrual debris from ectopic endometrium is probably not a major factor in the elicitation of the observed PF inflammation in infertile women with endometriosis and suggest an inverse relationship may exist between PF inflammation and the extent of endometriosis.     

Embryo quality before and after surgical treatment of endometriosis in infertile patients

Purpose

To investigate the hypothesis that surgical treatment of endometriosis in infertile patients may improve pregnancy rates by improving embryo quality.

Methods

We conducted a retrospective evaluation of 30 infertile patients treated with in vitro fertilization (IVF) before and after surgery for endometriosis. Patients served as their own controls and only cycles with similar stimulation protocols were compared.

Results

Using standard visual evaluation, embryo quality on day 3 was similar before and after surgical treatment of endometriosis. Fifty seven percent of patients had stage I–II endometriosis and 43% had stage III–IV disease. No patients had a live birth after the first IVF cycle and 43% of patients had a live birth with the IVF cycle after surgery.

Conclusions

Surgical treatment of endometriosis does not alter embryo quality in patients with infertility treated with IVF.