子宫内膜异位症患者腹腔液白细胞介素6,8及转化生长因子β1 …

OBJECTIVE: To investigate the role of cytokines in peritoneal fluid on pathogenesis of endometriosis (EM). METHODS: Interleukin-6 (IL-6), interleukin-8(IL-8) and transforming growth factor-beta 1 (TGF-beta 1) contents in peritoneal fluid (PF) of 31 cases with EM were detected by enzyme linked immunoabsorbent assay (ELISA) and compared with the counterparts of 22 cases without EM (controls). The correlation analyses between cytokine concentrations in peritoneal fluid of EM patients and the severity of EM or dysmenorrhea score were performed. RESULTS: The peritoneal fluid from patients with EM contained significantly greater amounts of IL-6 [(1.8 +/- 0.4) ng/L] and IL-8 [(1.7 +/- 0.5) ng/L] than those in controls [(1.2 +/- 0.2) ng/L and (1.4 +/- 0.3) ng/L respectively, P < 0.05]. However, in the amounts of TGF-beta 1 there were no significant difference (P > 0.05) between the two groups. The highest PF IL-6 and IL-8 concentrations were found in stage II, III and stag I, II EM respectively. A significant correlation between PF IL-6 content and the severity of disease was noted but there were no evidences of a relationship between concentrations of IL-8 and TGF-beta 1 and the severity of EM as well as between concentrations of three cytokines and dysmenorrhea score. CONCLUSION: Unusual levels of IL-6 and IL-8 in PF of EM patients partly account for imbalance of the immunologically dynamic environment in peritoneal cavity of EM patients.     

子宫内膜异位症患者辅助性T细胞亚群免疫状态的研究

目的　探讨辅助性T细胞 (Th)亚群在子宫内膜异位症 (内异症 )发病中的作用。方法　采用酶联免疫吸附法检测 30例内异症患者 (内异症组 )及 2 0例非内异症患者 (对照 1组 )血清及腹腔液中白细胞介素 (IL) 2、6的水平 ;用免疫组化技术分别检测IL 2、IL 6在内异症组患者异位内膜组织和 10例子宫肌瘤患者 (对照 2组 )的正常子宫内膜组织中的表达。结果　内异症组患者血清及腹腔液中位数IL 6水平分别为 5 3、2 1ng/L ,对照 1组患者血清及腹腔液中位数IL 6水平分别为 2 5、0 9ng/L ,两组妇女血清和腹腔液中IL 6水平分别比较 ,差异均有统计学意义 (P <0 0 5 ) ;内异症组Ⅲ～Ⅳ期患者血清及腹腔液中位数IL 6水平分别为 13 6、4 1ng/L ,Ⅰ～Ⅱ期患者血清及腹腔液中位数IL 6水平分别为 3 7、1 6ng/L ,Ⅲ～Ⅳ期患者与Ⅰ～Ⅱ期患者血清及腹腔液中位数IL 6水平比较 ,差异均有统计学意义 (P <0 0 5 ) ;内异症组IL 2 /IL 6比值在血清及腹腔液中分别为 0 7、1 1,均分别低于对照组的 0 8、6 2 ,差异也有统计学意义 (P <0 0 5 )。内异症组患者腹腔液与血清IL 6水平呈正相关 (r =0 74 5 ,P <0 0 1) ,血清及腹腔液中IL 6水平与IL 2 /IL 6比值均呈负相关 (r =- 0 4 0 6 ,P <0 0 5 ;r =- 0 4 80 ,P <0 0 5 )     

Expression of interleukin-1 (IL-1) beta messenger ribonucleic acid (mRNA) and IL-1 receptor antagonist mRNA in peritoneal macrophages from patients with endometriosis.

OBJECTIVE: To investigate the expression of interleukin-1 (IL-1) beta messenger ribonucleic acid (mRNA) and IL-1 receptor antagonist (IL-1ra) mRNA in peritoneal macrophages. DESIGN, SETTING: Peritoneal fluid (PF) samples were collected from patients who underwent laparoscopy or laparotomy. Northern blot analysis was performed at the reproductive research laboratory. PATIENTS: Twenty-six patients with endometriosis, 10 patients with postinflammatory pelvic adhesion, and 12 control women with normal pelvis. MAIN OUTCOME MEASURE: Polyadenylated RNA isolated from peritoneal macrophages was analyzed on Northern blots by using synthetic oligonucleotide probes. RESULTS: The level of IL-1 beta mRNA expression was elevated in the group with stage I endometriosis, whereas the increased expression of IL-1ra mRNA was observed in the group with stages III and IV endometriosis. The level of IL-1 beta mRNA showed a positive correlation with that of IL-1 beta in PF and a negative correlation with the level of IL-1ra mRNA. CONCLUSIONS: Our results suggest that peritoneal macrophages express IL-1ra mRNA rather than IL-1 beta mRNA with the progress of endometriosis and that peritoneal macrophages may secrete IL-1ra protein that modulates the effects of IL-1.     

Effect of interleukin-8 (IL-8), anti-IL-8, and IL-12 on endometrial cell survival in combined endometrial gland and stromal cell cultures derived from women with and without endometriosis.

OBJECTIVE: To study the affects of interleukin-8 (IL-8), anti-IL-8, and IL-12 on in vitro proliferation of endometrial cells. DESIGN: An in vitro study. SETTING: Department of Obstetrics and Gynecology, University of Aberdeen, UK. PATIENT(S): Women attending a fertility clinic. INTERVENTION(S): In vitro cell cultures using culture media supplemented with IL-8 (100 ng/mL, 200 ng/mL, and 500 ng/mL), IL-12 (1 ng/mL, 5 ng/mL, and 25 ng/mL), and anti-IL-8 (0.1 microg/mL, 1 microg/mL, 10 microg/mL). MAIN OUTCOME MEASURE(S): In vitro survival of dispersed endometrial cells (combined epithelial and glandular) at 5 and 9 days of culture. RESULT(S): There was a dose-dependent stimulatory effect of IL-8 on survival of cells. From women with and without endometriosis, IL-12 at 1 ng/mL significantly inhibited the survival of endometrial cells from women without endometriosis as compared with cells from women with endometriosis. At 1 microg/mL, anti-IL-8 significantly inhibited the survival of endometrial cells from women with endometriosis compared with cells from women without endometriosis on day 5 of culture. CONCLUSION(S): Our findings confirm the stimulatory effects of IL-8 and its possible role in the pathogenesis of endometriosis. The effects of IL-12 and anti-IL-8 on endometrial cell survival varied according to the disease state and the concentration of the cytokines. Future in vitro studies on the role of anti-IL-8 and IL-12 should aim to use a greater range of concentrations and a higher density of endometrial cells in cultures supplemented with monocytes.     

Angiogenic activity and IL-8 concentrations in peritoneal fluid and sera in endometriosis.

OBJECTIVES: During menstruation endometrial fragments are transported into the peritoneal cavity where they form endometriotic lesions. Angiogenesis is proposed as one of the mechanisms in endometriosis pathogenesis. The aim of the study was to determine the angiogenic activity and interleukin 8 concentrations in peritoneal fluid and sera in endometriosis. METHODS: Angiogenesis was determined in cutaneous assay in Balb/c mice; IL-8 concentrations were measured by ELISA test in sera and peritoneal fluid of 32 control and 56 endometriosis patients. Wilcoxon and Mann-Whitney tests and Spearman rank correlations were used in statistical analysis. RESULTS: Peritoneal fluid and sera from the examined group had higher angiogenic activity and interleukin 8 concentrations. There was correlation found between AFS and neovascularization induced by sera and PF of patients with peritoneal lesions. CONCLUSIONS: Angiogenesis plays an important role in pathogenesis of endometriosis. Although IL-8 takes part in neovascularization, there are other factors modulating angiogenesis in endometriosis.     

细胞间粘附分子-1在子宫内膜异位症在位内膜、腹腔液及异位灶中的表达及分析

目的:探讨ICAM 1与子宫内膜异位症的关系。方法:收集子宫内膜异位症患者(研究组)腹腔液30份、在位子宫内膜2 1份、异位病灶组织8份及非子宫内膜异位症患者中正常盆腔(对照组)腹腔液2 5份、子宫内膜2 0份。应用酶联免疫吸附试验双抗体夹心法测定腹腔液sICAM 1和免疫组化方法测定在位内膜、异位灶内膜ICAM 1表达。结果:子宫内膜异位症患者腹腔液中sICAM 1水平明显高于对照组(P <0 .0 1 ) ,子宫内膜的ICAM 1表达低于对照组同期子宫内膜的表达(P <0 .0 5 ) ,异位灶组织ICAM 1表达明显高于其在位内膜(P <0 .0 1 )及对照组内膜(P <0 .0 5 )。结论:子宫内膜异位症患者子宫内膜、腹腔液、异位灶内膜细胞间粘附分子的异常表达可能与子宫内膜异位症的发生发展关系密切。     

MMP-9及TIMP-1在子宫内膜异位症患者腹水及血清中的表达

目的:从蛋白水平检测内异症患者血清和腹水中MMP9及TIMP1浓度,探讨MMP9、TIMP1及腹水微环境在内异症发病中的作用。方法:收集47例RAFSI～IV期内异症患者的腹水,30例非内异症患者的腹水;从血清库中获取27例内异症患者血清,并收集26例正常妇女的血清作为对照。用ELISA法检测血清及腹水MMP9及TIMP1水平。结果:内异症患者腹水TIMP1与非内异症对照组无明显差异,但MMP9的水平明显高于对照组;内异症患者血清中MMP9的浓度与正常对照组差异有显著性,血清TIMP1的水平与对照组无明显差异,血清中MMP9/TIMP1的比值明显高于对照组;内异症患者血清MMP9浓度明显比腹水高,而TIMP1浓度明显比腹水低,因而血清中MMP9/TIMP1的比值是腹水的700倍左右。结论:MMP9及TIMP1参与内异症的发病,MMP9/TIMP1比值的测定具有临床意义;腹水微环境和内异症发生发展相互影响;血清的蛋白水解潜能远远大于腹水,其意义和解释需要进一步研究。     

深部浸润型与表浅型子宫内膜异位症者腹腔液Th1、Th2细胞因子变化

目的:研究深部浸润型与表浅型子宫内膜异位症(EMs)临床特征以及腹腔液Th1、Th2细胞因子的表达差异。方法:收集术后病理证实诊断为EMs患者的临床资料及其腹腔液,根据术中诊断分为:表浅型(腹膜型,n=30)、深部浸润型(n=16);以同期因输卵管因素不孕症行腹腔镜探查的妇女(n=16)为对照组。用ELISA法测定3组腹腔液中IL-2、IL-4、IL-6、IL-10、TNF-α、IFN-γ的表达差异性。结果:表浅型EMs患者年龄较轻、不孕率高、腹水发生率高,而性交痛则是深部浸润型EMs患者的典型临床特征。与对照组相比,EMs组腹腔液中IL-4、IL-6、IL-10、TNF-α表达均升高,IFN-γ在EMs组表达均降低。IL-2仅在深部浸润型EMs组升高;深部浸润型EMs组与表浅型EMs组之间对比,IL-6在表浅型EMs的升高明显(P<0.01),IL-10与TNF-α则在深部浸润型EMs组升高明显(P<0.01),IFN-γ在深部浸润型EMs组下降更为明显(P<0.01),IL-4在2个EMs组间表达无差异。结论:在表浅型与深部浸润型EMs的发病机制中,两者具有不同的免疫学改变。     

Concentrations of interleukin (IL)-1alpha, IL-1 soluble receptor type II (IL-1 sRII) and IL-1 receptor antagonist (IL-1 Ra) in the peritoneal fluid and serum of infertile women with endometriosis

OBJECTIVE: Endometriosis is an immune system-related gynaecological disease, characterised by an increase in number and activation of peritoneal macrophages. One of macrophage-derived factors is interleukin (IL)-1. The effects of IL-1 are inhibited by IL-1 receptor type II (IL-1 RII), soluble forms of IL-1 RII (IL-1 sRII) and IL-1 receptor antagonist (IL-1 Ra). The aim of our work was to study the IL-1alpha, IL-1 sRII and IL-1 Ra levels in the peritoneal fluid (PF) and serum of women with endometriosis in relation to stage of disease. STUDY DESIGN: Concentrations of IL-1alpha, IL-1 sRII and IL-1 Ra were measured by ELISA assay in the PF and serum of 58 women; 43 with and 15 without endometriosis (control group). RESULTS: Elevated PF and serum IL-1alpha and IL-1 Ra levels in the women with endometriosis in comparison with the control group were observed. IL-1 sRII levels in PF and serum were higher in the controls than in the women with endometriosis. Concentrations of IL-1alpha and IL-1 sRII were higher in advanced endometriosis, but higher IL-1 Ra was observed in the early stage of the disease. CONCLUSION: Impairment of regulation IL-1 activity in the peritoneal fluid and serum of women with endometriosis may play an important role in the pathogenesis and development of the disease.     

Soluble c-Met expression in the peritoneal fluid and serum of patients with different stages of endometriosis

Introduction

Hepatocyte growth factor (HGF), also known as scatter factor, and its receptor c-Met have been shown to be implicated in endometriosis. HGF acts as a mitogen, motogen, and morphogen on endometrial epithelial cells. The expression of c-Met on human endometrial cells has been reported. Many proteins are proteolytically released from the surface by a process known as ectodomain shedding. The aim of this study was to determine the levels of soluble c-Met (s-cMet) in the peritoneal fluid (PF) and serum samples of patients with different stages of endometriosis.

Material and methods

39 PF and serum samples from normal healthy and 130 samples from different stages of patients with endometriosis (33 cases of stage I, 38 stage II, 30 stage III and 29 stage IV) were included in this study. Total protein concentration (TPC) and the level of s-cMet in the PF and serum were determined by Bio-Rad protein assay based on the Bradford dye procedure and enzyme-linked immunosorbent assay, respectively.

Results

No significant change in the TPC was seen in the serum and PF of patients with endometriosis when compared with normal controls. Results obtained demonstrated that all PF and serum samples presented s-cMet expression, whereas, starting from stages I to IV endometriosis, a significant increase of s-cMet expression was observed as compared to controls.

Conclusion

The results of this study show that a high expression of s-cMet is correlated with advanced stages of endometriosis. It is also concluded that the detection of serum and PF s-cMet may be useful in classifying endometriosis.     

Differential flow cytometric detection of intracellular cytokines in peripheral and peritoneal mononuclear cells of women with endometriosis

OBJECTIVE: The pathogenesis of endometriosis is related to functional changes in CD3+ and CD14+ cells observed both at the local and systemic level. Here we investigated whether, and if so, how the body compartment influences cytokine expression in stimulated peritoneal and peripheral CD3+ and CD14+ cells of women with endometriosis. STUDY DESIGN: Isolated peripheral blood (PB) and peritoneal fluid (PF) mononuclear cells from women with endometriosis were cultured under non-adherent conditions and stimulated with PMA and ionomycin for 6h to induce intracellular cytokine synthesis of TNF-alpha, IFN-gamma, and IL-8 by CD3+ cells or with LPS for 9h to produce TNF-alpha, IL-6, IL-10, MCP-1, and IL-8 by CD14+ cells. RESULTS: The percentages of positive CD3+ cells stained for TNF-alpha and IFN-gamma were significantly higher and those stained for IL-8 were significantly lower in PF compared with PB, this being independent of the stage of endometriosis. In contrast, the percentages of CD14+ cells producing TNF-alpha, IL-6, IL-10, MCP-1, and IL-8 were significantly higher in PB than PF of women with endometriosis. CONCLUSIONS: Monocytes/macrophages and lymphocytes derived from the peripheral and peritoneal compartments of women with endometriosis differentially respond to stimulated cytokine synthesis induction. However, it is difficult to state whether the observed phenomenon is more related to body compartment influence per se or to the presence of endometriosis.     

Leptin is not involved in the pathophysiology of endometriosis-related infertility

OBJECTIVE: Changes in peritoneal fluid (PF) composition may affect fertilization as well as early embryonic development. Leptin, an adipocyte hormone, has been shown to act as a link between adipose tissue and the reproductive system. Therefore, we decided to assess peritoneal and serum leptin levels in infertile endometriotic patients. PATIENTS: Seventy-two women were studied, including 30 fertile and 18 infertile women with ovarian endometriotic cysts and, as a reference group, 24 patients with unexplained infertility. RESULTS: No significant difference in the peritoneal and leptin levels was found between the studied groups. Significantly higher PF leptin concentration was observed in patients with stages III and IV of endometriosis as compared to those with minimal stage of the disease. In fertile patients with endometriosis a positive correlation has been found between PF and serum leptin concentrations. CONCLUSIONS: No differences in peritoneal or serum leptin levels between infertile and fertile women with endometriosis suggest that this cytokine is not involved in pathophysiology of endometriosis-related infertility.     

Correlation of angiogenic cytokines-leptin and IL-8 in stage,type and presentation of endometriosis

Pelvic endometriosis is a chronic inflammatory disease with an immunological background. Yet there is paucity of contemporary research exploring both the angiogenic cytokines, leptin and IL-8 for a possible role in its pathophysiology. Objective: To compare levels of both leptin and IL-8 in peritoneal fluid (PF) in women with endometriosis vs. fertile controls and correlate with disease stage, type and symptoms. Materials and methods: PF from 58 women with endometriosis and 28 women undergoing tubal ligation was collected at laparoscopy and leptin and IL-8 levels were measured using ELISA. Results showed significantly higher levels of both cytokines in women with endometriosis. Significantly higher leptin and IL-8 levels were demonstrated in patients with early peritoneal (ASRM stage I and II) and advancing disease (ASRM stage III and IV), respectively. Levels of leptin/IL-8 were significantly lower in patients with endometrioma (4.8?ng/mL/32 pg/mL) vs. implants (13.0?ng/mL/68 pg/mL). There was no correlation of infertility or chronic pelvic pain with these levels. Conclusion: Both leptin and IL-8 levels are raised in PF of women with endometriosis reflecting inflammation and dysregulated immunomodulation. Higher levels of leptin were seen in early stages; IL-8 seems to stimulate the disease in a dose-dependent manner.     

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.     

Antigenic differences between the endometrium of women with and without endometriosis

Serum and peritoneal fluid from 12 women with endometriosis, 4 women with uterine leiomyomata and 6 fertile women without endometriosis (controls) and serum from 4 women with adenomyosis were tested with a passive hemagglutination assay for antibodies against endometrium from all the controls, 8 patients with endometriosis and all patients with uterine leiomyomata and from implants from 8 patients with endometriosis. Serum antibody titers in patients with endometriosis or leiomyomata were significantly higher against endometrial or implant antigens from patients with endometriosis and 2 patients with leiomyomata than those against the controls' endometrium. Peritoneal fluid endometrial antibody titers failed to reflect these antigenic differences. Controls and patients with adenomyosis had low titers of endometrial antibodies in their serum or peritoneal fluid. Antigenic differences appear to exist between the endometrium of patients with endometriosis and that of controls.