子宫内膜异位症患者腹腔液白细胞介素17水平的变化及其意义

目的探讨白细胞介素17(IL17)与子宫内膜异位症(内异症)发病的关系。方法应用酶联免疫吸附(ELISA)法检测36例不同期别内异症患者(内异症组)和26例非内异症患者(对照组)腹腔液中的IL17水平。结果内异症组和对照组腹腔液IL17水平分别为(5.7±1.9)ng/L和(5.3±1.4)ng/L,两组比较,差异无统计学意义(P>0.05)。按1985年美国生育学会内异症分期标准(rAFS)进行分期,Ⅰ～Ⅱ期内异症患者(16例)腹腔液IL17水平为(6.4±1.7)ng/L,Ⅲ～Ⅳ期内异症患者(20例)为(5.1±1.8)ng/L,两者比较,差异有统计学意义(P<0.05)。卵泡期和黄体期腹腔液IL17水平在两组间比较,差异均无统计学意义(P>0.05)。内异症不孕患者腹腔液IL17水平为(6.4±1.8)ng/L,内异症未合并不孕患者为(5.1±1.8)ng/L,两者比较,差异有统计学意义(P<0.05)。结论IL17水平变化可能与早期内异症及内异症不孕的发病有关。     

Different concentrations of interleukins in the peritoneal fluid of women with endometriosis: relationships with lymphocyte subsets.

The present study explored the possible relationships between immune cell subsets and interleukin (IL)-12 or IL-13 levels in the peritoneal fluid of patients with and without endometriosis. Peritoneal fluid samples were obtained from 80 women while they were undergoing laparoscopy for pain, infertility, tubal ligation or re-anastomosis. The American Fertility Society scoring system was used to determine the extension of endometriosis. The peritoneal fluid mononuclear cells were analyzed for immunophenotyping using cytometry, whereas peritoneal fluid concentrations of interleukins were measured using two ultrasensitive commercially available enzyme-linked imnunosorbent assay kits. Significantly higher peritoneal fluid IL-12 levels were found in women with moderate or severe endometriosis (stages III and IV) than in healthy controls (p < 0.01). Conversely, subjects with endometriosis showed remarkably lower peritoneal fluid IL-13 concentrations than controls, independent of the severity of the disease (p < 0.05). Considering immune system effectors, patients with endometriosis presented a significantly higher peritoneal fluid CD8+/CD4+ ratio when compared with healthy controls. Moreover, the number of peritoneal fluid CD8+ and CD4+ activated T cells was significantly lower in the former than in the latter group, independent of the endometriosis stage. Connections were observed between peritoneal fluid interleukins and peritoneal fluid T cells: both patients with endometriosis and controls presented an inverse correlation between peritoneal fluid activated T cells and IL-13 levels, and a direct correlation between peritoneal fluid T cells and IL-12 concentrations. These data seem to suggest that a reciprocal modulation exists between peritoneal fluid cytokines and T lymphocyte subsets in patients with endometriosis.     

The evaluation of IL-12 levels in peritoneal fluid and serum of women with endometriosis

We studied levels of IL-12 in peritoneal fluid and serum in patients with minimal, advanced and recurrent endometriosis compared to women without endometriotic lesions in pelvis minor. The aim of the study was to determine whether level of IL-12 detected in peritoneal fluid or serum changes with grading of severity of endometriosis. To assess IL-12 levels immunosorbent ELISA was used. There were no statistically significant differences in IL-12 levels in peritoneal fluid nor in serum in any of studied groups. There was higher concentration (without statistical significance) of IL-12 in peritoneal fluid of healthy women compared to women with endometriosis.     

T helper (Th)1, Th2, and Th17 interleukin pathways in infertile patients with minimal/mild endometriosis

In the present study, interleukin (IL)-10, IL-12, IL-17, and IL-23 levels were measured in serum and peritoneal fluid of women with minimal or mild endometriosis and compared with levels in controls without endometriosis. Higher IL-23 levels were encountered in the peritoneal fluid of women with endometriosis, suggesting a possible role of this cytokine in these women's infertility.     

Matrix metalloproteinase 2 is associated with changes in steroid hormones in the sera and peritoneal fluid of patients with endometriosis

OBJECTIVE: To determine matrix metalloproteinase 2 (MMP-2) levels in sera and peritoneal fluids of women with or without endometriosis-associated infertility and to determine whether MMP-2 had correlation with 17beta-E(2) and P in the sera and peritoneal fluids of the patients with endometriosis. DESIGN: A case-control study. SETTING: A women's hospital in China. PATIENT(S): Forty patients with endometriosis-associated infertility undergoing laparoscopy and 18 women undergoing tubal ligation. INTERVENTION(S): Sera and peritoneal fluids were collected from women with or without endometriosis. MAIN OUTCOME MEASURE(S): Matrix metalloproteinase 2, 17beta-E(2), and P concentration. RESULT(S): Compared with the control women, the patients with endometriosis had higher MMP-2 levels in their sera and peritoneal fluids. Serum and peritoneal fluid MMP-2 levels were higher in the proliferative phase than in the secretory phase. The level of MMP-2 in the patients was correlated positively with 17beta-E(2) in serum and peritoneal fluid and negatively with P. CONCLUSION(S): The results demonstrated a correlation between MMP-2 concentrations and steroid hormones in sera and peritoneal fluids of the patients with endometriosis-associated infertility. Estradiol might up-regulate MMP-2, resulting in the formation of endometriosis. Progesterone might down-regulate MMP-2 to inhibit the formation and development of endometriosis.     

The role of the peritoneal macrophage in infertility associated with endometriosis--animal experiment

In this study, the role of the peritoneal macrophage (M phi) in the infertility of endometriosis patients was investigated through clinical and animal experiments. In addition, the effect of prednisolone as a treatment of infertility caused by endometriosis was evaluated in animal experiments. The number of M phi and the level of IL-1 in the peritoneal fluid were measured. The results showed that the number of M phi and the level of IL-1 increased in 1/3 of patients with endometriosis. Next, the effect of M phi and IL-1 on the pregnancy rate and the number of conceptions was studied in animal experiments. The conception rate and littler number significantly decreased when M phi increased in the animal peritoneal cavity after injection of OK-432. In the experiment with IL-1 injection into the peritoneal cavity, similar results were obtained, and in the group of animal with suppression of M phi function due to prednisolone administration, both of the rate of pregnancy and the number of litters increased remarkably. It can therefore be assumed that the mechanism of infertility in women with endometriosis is associated with peritoneal M phi, or, more specifically, with M phi-produced IL-1.     

Chemokine growth-regulated-alpha: a possible role in the pathogenesis of endometriosis.

Endometriosis is a common disease in women at reproductive age. We investigated the concentration of neutrophil-activating factor (growth-regulated gene-alpha; GRO-alpha) (a member of the chemokine family), in peritoneal fluid of infertile women with or without endometriosis. Peritoneal fluid was obtained laparoscopically from 22 women with and 21 without visible endometriotic lesions. GRO-alpha concentration was measured by the use of an ELISA kit. Median concentration of GRO-alpha was 87.65 +/- 56.19 pg/ml in the study group and 60.72 +/- 11.98 pg/ml in the control group. The distribution of data differed from normal, therefore logarithmic transformation of data was performed. Concentration of GRO-alpha was significantly higher in the peritoneal fluid of women with endometriosis when compared with controls (p = 0.05). No correlation between concentration of GRO-alpha, stage of endometriosis, duration of infertility or sex steroid hormone levels was found. The study shows that GRO-alpha may play a role in the pathogenesis of endometriosis, possibly by chemoattraction and activation of neutrophils present in higher numbers in the peritoneal fluid of women with endometriosis. It is also feasible that the angiogenic properties of GRO-alpha might prompt the progression of endometriotic lesions.     

The concentration of 13,14-dihydro-15-keto prostaglandin F2 alpha and prostaglandin E2 in peritoneal fluid of infertile patients with and without endometriosis

Prostaglandin F2 alpha (PGF2 alpha) metabolite (13,14-dihydro-15-keto PGF2 alpha) and prostaglandin E2 (PGE2) were assayed in the cul-de-sac fluid aspirated from 15 patients with endometriosis, in saline peritoneal washings of 5 patients with unexplained infertility, and in 5 control subjects. The fluid from the cul-de-sac of patients with endometriosis showed wide variations in the concentrations of PGF2 alpha metabolite and PGE2. There was no correlation between the concentration of these prostaglandins (PGs) and the stage of the cycle. Saline peritoneal washings from patients with unexplained infertility had significantly higher concentrations of PGF2 alpha metabolite than the control subjects. The concentration of PGE2 in the saline peritoneal washings was higher in the group with unexplained infertility than in the control group; however, the difference was not significant. The concentration of PGF2 alpha metabolite and PGE2 in the peritoneal saline washings from patients with unexplained infertility were no different from the concentrations of these PGs in the peritoneal fluid (PF) from patients with endometriosis. PG concentration in PF of infertile patients with or without endometriosis is a new variable for evaluation in these patients.     

Effect of ovarian involvement on peritoneal fluid cytokine concentrations in endometriosis patients

Peritoneal fluid cytokines are important for initiation and progression of endometriosis. The objective of this study was to compare a group of five cytokines (interleukins IL-1 beta, IL-6, IL-8, IL-13 and tumour necrosis factor alpha; TNFalpha) in peritoneal fluid of endometriosis patients with ovarian involvement (Group I, n = 17) to those in patients without ovarian involvement (Group II, n = 33) and to a reference group without endometriosis (Group III, n = 25). All three groups were comparable regarding age, parity and body mass index. IL-8 concentrations were significantly higher in groups I and II compared with the reference group (P = 0.01 and 0.02, respectively). Similarly, TNFalpha concentrations were significantly higher in groups I and II compared with the reference group (P < 0.0001 and 0.0004, respectively). All other cytokines were comparable in the three groups. No significant differences were found between groups I and II with respect to the cytokines measured. In conclusion, peritoneal fluid IL-8 and TNFalpha concentrations are significantly higher in endometriosis. Ovarian involvement does not alter the pattern of cytokines. It appears that the inflammatory mediators of endometriosis are similar with and without ovarian involvement.     

Altered expression of interleukin-18 in the peritoneal fluid of women with endometriosis

OBJECTIVE: Peritoneal fluid (PF) inflammatory factors may participate in the pathogenesis of endometriosis. The aim of this study was to investigate PF interleukin (IL)-18 levels in women with and without endometriosis. DESIGN: Controlled clinical study. SETTING: Women undergoing laparoscopy at a university hospital. PATIENT(S): Fifty women with previously untreated endometriosis, 8 women on GnRH agonists for endometriosis, and 18 control women with normal pelvic anatomy who were undergoing tubal ligation. INTERVENTION(S): Peritoneal fluid IL-18 levels as measured by ELISA. MAIN OUTCOME MEASURE(S): Peritoneal fluid IL-18 levels. RESULT(S): Peritoneal fluid IL-18 levels were significantly higher in women with previously untreated endometriosis (mean +/- SEM, 91.1 +/- 6.5 pg/mL) than in control women (59.4 +/- 2.0 pg/mL). Interestingly, women with superficial (100.0 +/- 10.2 pg/mL) and deep peritoneal implants (94.0 +/- 10.8 pg/mL) had significantly higher PF IL-18 levels than did women with endometriomas (57.8 +/- 1.8 pg/mL). Similarly, women with stage I-II endometriosis (97.3 +/- 8.0 pg/mL), but not women with stage III-IV endometriosis (74.9 +/- 9.9 pg/mL), had significantly higher PF IL-18 levels than did control women. Peritoneal fluid IL-18 levels were significantly higher in the luteal phase than in the follicular phase but did not discriminate between women with pelvic pain or infertility. CONCLUSION(S): Peritoneal fluid IL-18 is elevated in women with peritoneal, minimal- to mild-stage endometriosis.     

Peritoneal fluid leptin concentration in infertile patients.

Leptin may play a role in the regulation of menstrual cycle acting either directly on ovaries or at the level of the hypothalamic-pituitary axis. Peritoneal fluid is a biologically active environment that influences ovarian function but, on the other hand, concentration of many substances in peritoneal fluid can reflect the functional status of ovaries. In our study, we estimated leptin concentration in peritoneal fluid in relation to serum leptin concentration in infertile patients. A study group consisted of 31 infertile patients that underwent laparoscopy. In 15 patients, laparoscopy did not solve the problem and they were grouped as unexplained infertility, in eight patients we found endometriosis and eight patients were diagnosed as polycystic ovary syndrome (PCOS). We found significantly higher peritoneal fluid leptin concentrations in patients with unexplained infertility and endometriosis compared to those with PCOS. The plasma leptin concentration did not differ between the groups studied. There was no correlation between peritoneal fluid and plasma leptin concentrations in any patient group. Taking into account our results, the role of leptin in reproduction should be emphasized but further investigation is needed to determine its mechanism of action.     

Cytokine profiles in autologous peritoneal fluid and peripheral blood of women with deep and superficial endometriosis

In a preliminary study the hypothesis was tested that cytokine profiles in peripheral blood were higher in women with deep infiltrating endometriosis and cytokine profiles in peritoneal fluid were higher in women with superficial endometriosis. Thirteen women of reproductive age having laparoscopy for infertility (n=9), pain (n=3) or combined pain and infertility (n=1). Peripheral blood and peritoneal fluid were obtained and analyzed for Interleukin-6 (IL-6), Tumor Necrosis Factor-alpha (TNF-α), Interleukin-10 (IL-10), Transforming Growth Factor-betal (TGFβ1), and Interferon-gamma (IFN-γ). No significant cytokine differences were observed in either peritoneal fluid or peripheral blood between IL-6, TGFβ1, IFNγ, TNF-alpha and IL-10 of women with superficial endometriosis (n=7) and women with deeply infiltrating endometriosis (n=6). The results of this preliminary study do not show significant differences in peripheral blood and peritoneal fluid cytokine levels between women with deep infiltrating endometriosis compared to women with superficial disease. Future studies with increased sample size are required to either confirm or refute these preliminary findings. Received: 5 January 2000 / Accepted: 14 August 2000     

Role of cytokines in endometriosis-associated infertility

Endometriosis, which is common in women of reproductive age, may affect fertility. It is also clear that mechanical disruption of the pelvic anatomy may cause infertility. However, our understanding of the association between the early stage of endometriosis and infertility remains incomplete. Bloody peritoneal fluid (PF) is frequently observed in the cul-de-sac of endometriosis patients and contains various biologically active factors. We found that the concentrations of tumor necrosis factor alpha (TNF-alpha) and interleukin-6 (IL-6) in PF from patients with endometriosis were significantly higher than that of patients with endometriosis. There were significantly positive correlations between the levels of TNF-alpha and IL-6. We compared the levels of these cytokines with regard to the R-AFS stages and scores, but no differences were observed. In contrast, these cytokines correlate with the number and extent of red color peritoneal endometriosis. TNF-alpha increased the expression of IL-6 messenger RNA and protein in endometriotic stromal cells derived from chocolate cyst in a dose-dependent manner. The addition of IL-6 inhibited the development of mouse preimplantation embryo and impaired sperm function. We concluded that increased levels of IL-6 in peritoneal fluid of patients with active red endometriosis might be related to endometriosis-associated fertility.     

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.     

VEGF, TNF-alpha and IL-6 in the peritoneal fluid of women with unexplained infertility

Unexplained infertility is an important problem in diagnosis and therapy in everyday gynecologist's practice. Looking for possible reasons of the unexplained infertility we studied the concentrations of selected cytokines (VEGF, TNF-alpha and IL-6) in the peritoneal fluid of women suffering from the unexplained infertility. We compared the results in the studied group with the control group and with the patients with endometriosis. Immunological disorders of the peritoneal fluid in endometriosis are thought to take part in its pathomechanism. Our results suggest that the levels of one of the main factors of endothelium proliferation (VEGF) in the peritoneal fluid from women with unexplained infertility and women with endometriosis are comparable. Concentration IL-6 and TNF-alpha in the peritoneal fluid in case of unexplained infertility and control group was lower than in the endometriosis patients.     

Peritoneal Fluid Biochemistry in Infertile Women with Mild Pelvic Endometriosis

Although the factors involved in the pathophysiology of endometriosis are probably multiple and interrelated, prostaglandins may play an important role in the infertility of women with mild disease. In the present study, prostaglandin F2 alpha (PGF2 alpha) and 17 beta-oestradiol were measured in the peritoneal fluid of a selected group of infertile women who had mild pelvic endometriosis (without anatomical distortion) and compared with those values in normal women who had no pelvic disease and in women with pelvic infection. Although there was a wide scatter of PGF2 alpha values, the mean (1,066 pg/ml) in the endometriosis group was significantly greater than that in the other 2 groups (542 pg/ml, normal and 688 pg/ml, pelvic infection); the increase was found in both phases of the menstrual cycle. The mean concentration of 17 beta oestradiol was markedly higher in the luteal than the follicular phase in all 3 groups; however, no significant differences were found between the groups. Interestingly, the mean value of PGF2 alpha and 17 beta-oestradiol was higher in women with endometriosis who failed to conceive than in those who became pregnant. An estimation of PGF2 alpha in the peritoneal fluid may have prognostic value in the evaluation of infertile patients, especially those with mild endometriosis or in whom the problem is unexplained.     

Chemokine growth-regulated-α: a possible role in the pathogenesis of endometriosis

Endometriosis is a common disease in women at reproductive age. We investigated the concentration of neutrophil-activating factor (growth-regulated gene-α; GRO-α) (a member of the chemokine family), in peritoneal fluid of infertile women with or without endometriosis. Peritoneal fluid was obtained laparoscopically from 22 women with and 21 without visible endometriotic lesions. GRO-α concentration was measured by the use of an ELISA kit. Median concentration of GRO-α was 87.65 ± 56.19 pg/ml in the study group and 60.72 ± 11.98 pg/ml in the control group. The distribution of data differed from normal, therefore logarithmic transformation of data was performed. Concentration of GRO-α was significantly higher in the peritoneal fluid of women with endometriosis when compared with controls (p = 0.05). No correlation between concentration of GRO-α, stage of endometriosis, duration of infertility or sex steroid hormone levels was found. The study shows that GRO-α may play a role in the pathogenesis of endometriosis, possibly by chemoattraction and activation of neutrophils present in higher numbers in the peritoneal fluid of women with endometriosis. It is also feasible that the angiogenic properties of GRO-α might prompt the progression of endometriotic lesions.     

Increased nitric oxide in peritoneal fluid from women with idiopathic infertility and endometriosis.

OBJECTIVE: To verify whether nitric oxide in peritoneal fluid is associated with endometriosis and infertility. STUDY DESIGN: Twenty-five women with idiopathic infertility and 38 with endometriosis were recruited, and 18 cases of uterine myomata and 2 cases of ovarian cyst served as controls. Peritoneal fluid samples were aspirated from the pouch of Douglas during laparoscopy or laparotomy. Metabolites of nitric oxide (nitrite and nitrate) in peritoneal fluid were determined by a method using nitrate reductase and the Griess reaction. RESULTS: Peritoneal concentrations of nitrate/nitrite in both infertile women (42.02 +/- 12.98 mmol/L) and patients with endometriosis (41.75 +/- 16.42 mmol/L) were significantly higher than that in controls (33.96 +/- 13.07, P < .05 for both). No significant difference in peritoneal nitrate/nitrite level was found between infertile women and patients with endometriosis (P > .5). Peritoneal levels of nitrate/nitrite were comparable among patients with endometriosis at different stages (P > .5). Patients with endometriosis had more peritoneal fluid than controls and idiopathic infertile women, while controls and idiopathic infertile women had comparable amounts of peritoneal fluid. CONCLUSION: An increased peritoneal level of nitric oxide is a common alteration in endometriosis, endometriosis-associated infertility and idiopathic infertility and may be associated with the pathogenesis of these diseases.     

Decreased peritoneal concentrations of interleukin-15 in women with advanced stage endometriosis

OBJECTIVE: To assess the concentrations of interleukin-15 (IL-15) in peritoneal fluid from women with endometriosis and fertile disease-free controls. STUDY DESIGN: Peritoneal fluid samples were obtained from 50 women with endometriosis and 29 fertile women having tubal ligation. Concentrations of IL-15 were measured. RESULTS: The mean (S.D.s) concentration of IL-15 in peritoneal fluid was 11.17 pg/mL (3.89) for women with endometriosis, and 12.59 pg/mL (4.11) for fertile disease-free controls. The difference of peritoneal IL-15 concentrations between endometriosis and control women was not statistically significant. However, peritoneal IL-15 concentrations were significantly lower in women with moderate/severe endometriosis when compared with those in women with minimal/mild endometriosis, and in controls (P<0.05). In addition, peritoneal IL-15 concentrations did not correlate with the phase of menstrual cycle in endometriosis or control women. CONCLUSIONS: Our results suggest that the decreased peritoneal IL-15 concentrations in women with moderate/severe endometriosis imply a role of IL-15 in the pathogenesis of advanced endometriosis as compared to those with minimal/mild endometriosis and fertile disease-free controls.     

High concentrations of the CA-125, CA 19-9 and CA 15-3 in the peritoneal fluid between patients with and without endometriosis

Objective In the present study we compared the levels of CA-125, CA 19-9, and CA 15-3 in the peritoneal fluid (PF) of patients with and without endometriosis, then assessed the possibility of a correlation among these tumor markers.Study design Our study was a controlled clinical study of patients undergoing laparoscopy for infertility or other benign gynecology conditions. Peritoneal fluid samples were collected from 65 women with endometriosis and 43 women without pelvic disease. Levels of CA-125, CA 19-9 and CA 15-3 in the peritoneal fluid were determined by immunoradiometric assay.Results The concentration of CA-125 in PF from patients with endometriosis was significantly higher than that in the control group (p<0.001); for CA 19-9 and CA 15-3, PF concentrations were not statistically different between these two groups. Women with endometriosis had significantly higher levels of CA-125 in proliferative and secretory phases than the control group (p<0.001 and p<0.002 respectively); furthermore, in patients with endometriosis the CA 19-9 levels were significantly lower in secretory phase than the proliferative (p<0.004). The levels of CA-125 were significantly lower in women with tubal ligation, in comparison with infertility or pelvic pain in the control group (p<0.001). No significant difference was seen in women with infertility or pelvic pain in endometriosis group and the levels of CA-125, CA 19-9, and CA 15-3. We did not find any correlation between the stages of endometriosis and the concentration of CA-125, CA 19-9 and CA 15-3. A significant correlation between the CA 19-9 levels and CA 15-3 in patients with endometriosis was found (r=0.72, p=0.001).Conclusions We found high concentrations of CA-125, CA 19-9, and CA 15-3 in the peritoneal fluid of women with and without endometriosis in the Yale series. However, the levels only of CA-125 were higher in women with endometriosis, but without diagnostic value. The role of simultaneously high concentrations of CA 19-9 and CA 15-3 in women with endometriosis needs to be explored further.