腹腔液及异位内膜组织MCP-1、IGF-Ⅰ水平变化与子宫内膜异位症发病及不孕的相关性分析

目的探讨子宫内膜异位症（Endometriosis，EM）患者腹腔液、异位内膜组织中单核细胞趋化蛋白-1（monocyte chemotactic protein-1，MCP-1）、胰岛素样生长因子-1（Insulin-like growth factors-Ⅰ，IGF-Ⅰ）的变化在子宫内膜异住症发病及不孕中的作用。方法EM患者30例作为观察组，其中早期病例（Ⅰ-Ⅱ期）10例，晚期病例（Ⅲ-Ⅳ）20例；合并不孕症患者12例，非不孕症者18例。10例无内膜异位症者作为对照组。用酶联免疫检测（ELISA）方法测定两组腹腔液、内膜组织中的MCP-1、IGF-Ⅰ含量的变化。结果（1）内膜异位症组腹腔液、异位内膜组织中MCP-1、IGF-Ⅰ水平与对照组相比较有显著差异（P〈0．01），且早期病例腹腔液MCP-1、IGF-Ⅰ水平及内膜组织MCP-1水平均明显高于晚期病例，差异显著（P〈0．05）。（2）内异症合并不孕症患者腹腔液IGF-Ⅰ水平与非不孕症比较有显著差异（P〈0．05）。结论内膜异位症患者腹腔液中MCP-1、IGF-Ⅰ的变化在子宫内膜异位症发病机制中具有一定作用。     

IL-12p70、p40在子宫内膜异位症患者腹腔液中的表达及意义

目的　通过测定子宫内膜异位症患者腹腔液中IL - 12 p70及IL - 12p4 0的水平 ,探讨其与子宫内膜异位症发生发展的关系。方法　采用酶联免疫吸附法 (ELISA)测定 2 0例子宫内膜异位症患者 (内异症组 )及 2 0例卵巢良性肿瘤患者(对照组 )腹腔液中IL - 12p70、IL - 12 p4 0的水平。比较两组间两个细胞因子水平以探讨它们与内异症发生的关系 ,并探讨其是否与疾病的进展有关。结果　腹腔液中IL - 12 p70水平内异症组稍低于对照组 ,但两者之间差异无显著性 (P >0 0 5 ) ;IL- 12 p4 0水平内异症组明显高于对照组 ,差异有显著性 (P <0 0 5 ) ;内异症组中 (Ⅰ～Ⅱ期 )患者IL - 12 p4 0水平较 (Ⅲ～Ⅳ期 )患者稍高 ,但两者差异并无显著性 (P >0 0 5 ) ;而 (Ⅰ～Ⅱ期 )患者IL - 12 p70水平明显高于 (Ⅲ～Ⅳ期 )患者 ,差异有显著性 (P <0 0 5 )。结论　内异症患者腹腔液中IL - 12p4 0水平的增高可能是自然杀伤 (NK)细胞功能受损的部分原因 ,从而可能与子宫内膜异位症的发生有一定关系 ;腹腔液中IL - 12 p70水平随内异症疾病程度进展而下降 ,表明其可能是内异症中NK细胞功能进一步下降的原因之一 ,从而可能是促使内异症进一步发展的原因之一。     

子宫内膜异位症患者端粒酶及端粒酶催化亚单位的表达研究

目的探索研究不同分型的子宫内膜异位症患者体内腹腔液中端粒酶及端粒酶逆转录酶的表达及对子宫内膜异位症发展的影响。方法在腹腔镜下收集64例子宫内膜异位症手术患者体内腹腔液,应用酶联免疫吸附实验（ELISA）及荧光原住杂交实验（FISH）方法分别检验收集的腹腔液中端粒酶的阳性表达及端粒酶逆转录酶（hTERC）的扩增情况,并与对照组（盆腔炎患者）相比较。结果应用ELISA方法检验出子宫内膜异位症组腹腔液中端粒酶阳性率与对照组差异无统计学意义;FISH实验检验出子宫内膜异位症组腹腔液中人类端粒酶逆转录酶扩增阳性率0-Ⅱ期与对照组差异无统计学意义;Ⅲ-Ⅳ期患者于对照组差异有统计学意义。结论子宫内膜异位症患者腹腔液中端粒酶逆转录酶表达均高于实验对照组,其可能与异位子宫内膜的恶变及增值活性较高相关。     

腹膜间皮细胞在小鼠子宫内膜细胞刺激腹腔微环境中作用

目的：探讨腹腔微环境受子宫内膜细胞刺激改变中腹膜间皮细胞的作用及其对子宫内膜异位症发病机制的意义。方法： 注射小鼠子宫内膜上皮和间质细胞入小鼠腹腔，酶联免疫吸附法（ELISA）检测注射后4、24和72 h时点腹腔冲洗液细胞因子MCP-1/JE、IL-1α和IL-6蛋白表达，同步逆转录-聚合酶链式反应技术 (Real-Time RT-PCR) 检测腹膜组织 (主要含腹膜间皮细胞) 和腹腔巨噬细胞的细胞因子MCP-1/JE、IL-1 α和IL-6 mRNA表达。结果: 子宫内膜细胞刺激腹腔细胞因子蛋白含量迅速一过性升高，4 h时点为表达高峰，腹膜细胞因子基因表达与腹腔液蛋白表达同步，腹腔巨噬细胞基因表达高峰滞后于腹腔液蛋白表达。子宫内膜上皮细胞刺激腹腔炎症反应作用强于间质细胞。结论: 子宫内膜细胞刺激腹腔发生非特异性炎症反应，腹膜间皮细胞可能是其细胞因子效应的主要来源，提示腹膜在子宫内膜异位症中除作为病灶依附体外，还可能在腹腔微环境无菌性炎症反应中起重要作用。     

子宫内膜异位症患者腹腔液白介素6浓度的Meta分析

为了确定子宫内膜异位症（EM）患者腹腔液白介素6（IL－6）浓度是否高于正常盆腔者（对照组），本文采用Meta分析，对有关EM患者腹腔液IL－6浓度报道的国内外文献资料进行定量综合分析。结果提示：按ELISA法EM组腹腔液IL－6的水平较对照组高出（－0．472－3．974）倍标准差。无论采取方法的不同，高出（－0．73－3．45）倍标准差，均提示不能笼统认为EM患者腹腔液IL－6浓度较对照组高；但对按照生育协会（AFS）校正标准将EM分期进行分组：I－Ⅱ期归为一组（EMI组），Ⅲ-Ⅳ期归为一组（EM2组）的资料，经Meta分析后，发现EM患者腹腔液IL－6升高水平与EM的病情严重程度有较好的相关性：其中EMI组较对照组高出（－4．807－5．608）倍标准差，两组间无差异；但EM2组较EM1组和对照组分别高出（0．54－5．34），（1．13－5．46）倍标准差，明显高于后两组。EM患者腹腔液IL－6浓度随EM病变程度而升高，IL－6在EM的发生发展中具有一定的病理生理学意义。     

子宫内膜异位症患者NK细胞和γδT淋巴细胞分析

目的：探讨子宫内膜异位症患者外周血、腹腔液中NK细胞和γδT淋巴细胞的数量和杀伤功能。方法：取15例子宫内膜异位症患者的外周血和腹腔液,用荧光染色法、流式细胞术和酶标法等测定NK和γδT细胞的数量和杀伤功能。取10例非内异症者外周血和腹腔液作为对照。结果：1．与对照组相比,病例组外周血和腹腔液中的NK细胞数量无明显改变,γδT细胞数量在外周血中无明显差异,但在腹腔液中明显增加（P＜0．05）。病例组中外周血和腹腔液之间NK和γδT细胞数量均有差异,表现为腹腔液中两种细胞所占的百分比明显增高（P＜0．05）。2．病例组与对照组相比,除外周血的γδT细胞外,外周血的NK细胞及腹腔液的NK和γδT细胞的功能均明显下降（P＜0．05）。同一个体中,病例组腹腔液中的NK和γδT细胞的功能比外周血中的明显降低（P均＜0．05）。结论：内异症患者NK和γδT细胞杀伤功能的下降可导致机体清除异位子宫内膜细胞的功能下降,其改变可能在子宫内膜异位症发病过程中起一定的作用。     

子宫内膜异位症患者瘦素检测及临床意义

目的检测子宫内膜异位症(EMS)患者血清和腹腔液瘦素(Leptin)水平的变化及在子宫内膜表达情况,探讨其在内异症发病中的作用。方法放射免疫法测定40例内异症患者和40例非内异症患者妇女(对照组1)血清和腹腔液Leptin的水平。抗生物素蛋白-过氧化物酶(SP)法检测瘦素在子宫内膜表达情况,并与R-AFS分期进行相关分析。同期检测50例健康妇女(对照组2)血清Leptin水平。结果 EMS患者血清瘦素水平与对照组间比较无差异;腹腔液中瘦素水平明显高于对照组;EMS患者和对照组子宫内膜中均有瘦素表达,EMS患者与对照组表达上存在差异(P<0.05)。结论内异症患者腹腔液中瘦素水平明显升高,并随疾病的严重程度而改变,同期影响瘦素在子宫内膜中的表达,这可能是瘦素在内异症的发生发展中起作用的机制。     

子宫内膜异位症患者血清与腹腔液中骨桥蛋白的表达及意义

目的探讨骨桥蛋白(OPN)在子宫内膜异位症患者血清与腹腔液中的表达及意义。方法用酶联免疫吸附方法检测40例子宫内膜异位症患者术前血清与腹腔液中骨桥蛋白的表达情况。结果 (1)子宫内膜异位症患者术前血清OPN平均水平为(46.68±16.38)ng/ml(16.69~101.85ng/ml),明显高于对照组患者水平(P<0.05);内异症组患者的腹腔液OPN平均水平为(47.47±15.90)ng/ml(20.35~97.56ng/ml),也高于对照组患者的表达水平(P<0.05)。(2)Ⅲ~Ⅳ期内异症患者的血清与腹腔液中OPN的表达水平均高于I~Ⅱ期患者,差异具有显著性(P<0.05)。(3)内异症患者的血清与腹腔液的OPN表达呈正相关关系(r=0.669,P<0.05);对照组血清与腹腔液的0PN表达亦呈正相关关系(r=0.538,P<0.05)。结论 OPN在子宫内膜异位症患者血清与腹腔液中高表达,可能促进异位内膜黏附和侵袭,促进血管生成,在子宫内膜异位症发生发展中起重要作用,OPN在内异症的早期诊断方面有一定应用价值。     

基质金属蛋白酶及其抑制剂在子宫内膜异位症中的研究进展

经血逆流是腹腔子宫内膜异位症的主要发病机制之一,子宫内膜植入其他组织的过程需要细胞外基质的降解,基质金属蛋白酶是降解细胞外基质的重要酶类。近年来研究表明,基质金属蛋白酶及其抑制剂的异常表达在子宫内膜异位症的发生和发展过程中起着重要作用。     

基质金属蛋白酶及其抑制剂在子宫内膜异位症中的研究进展

经血逆流是腹腔子宫内膜异位症的主要发病机制之一，子宫内膜植入其他组织的过程需要细胞外基质的降解，基质金属蛋白酶是降解细胞外基质的重要酶类。近年来研究表明，基质金属蛋白酶及其抑制剂的异常表达在子宫内膜异位症的发生和发展过程中起着重要作用。     

Decreased levels of the potent regulator of monocyte/macrophage activation, interleukin-13, in the peritoneal fluid of patients with endometriosis

Endometriosis is characterized by an increase in the number, activation and secretory activity of peritoneal fluid macrophages. Factors regulating the activation of these cells may be important in the pathophysiology of this disease. In this study we measured by enzyme- linked immunosorbent assay the concentrations of the macrophage inhibitory factor interleukin (IL)-13 in the peritoneal fluid of women with and without endometriosis. It was found that women with endometriosis had significantly lower amounts of IL-13 (95 +/- 9.8 pg/ml) in peritoneal fluid, compared with women without endometriosis (115 +/- 30 pg/ml) (P < 0.01). No cycle-specific variation was evident for either group. Another macrophage inhibitory interleukin (IL-10) was also measured, but no differences between women with (16.1 +/- 13.2 pg/ml) or without (10.3 +/- 5.6 pg/ml) endometriosis were seen. The immunolocalization of IL-13 was assessed in eutopic and ectopic endometrium and in isolated peritoneal fluid cells. Glandular epithelial cells and stromal cells in both eutopic and ectopic endometrium were immunopositive for IL-13. No cycle-specific differences in the immunolocalization of IL-13 were seen. In conclusion, the reduced amounts of IL-13 in the peritoneal fluid of women with endometriosis may lead to a lack of suppression of macrophage activation, thereby contributing to the overall pathogenesis of this disease.      

Peritoneal fluid concentrations of interleukin-8 in women with endometriosis: relationship to stage of disease

There is increasing evidence that immunological mechanisms play a role in the pathogenesis and pathophysiology of endometriosis. It was therefore of interest to study interleukin-8 (IL-8), a chemokine, in the peritoneal fluid and peripheral blood of women undergoing laparoscopic procedures. The presence and concentrations of IL-8 in relation to endometriosis, infertility and abdominal pain were evaluated. Samples of peritoneal fluid (n = 49) and peripheral blood (n = 50) were obtained from 50 consecutive patients undergoing laparoscopic surgery for various gynaecological indications (abdominal pain, infertility, sterilization). IL-8 was present in the peritoneal fluid of most women (87%). The concentration of IL-8 in the peritoneal fluid was higher in women with endometriosis compared to women without (P = 0.02). This difference was more pronounced in early (stage 1) endometriosis (P = 0.001). IL-8 concentrations in the peritoneal fluid were also higher in women with early endometriosis compared to women with later stages of the disease (P = 0.003). Peripheral blood concentrations did not correlate with peritoneal fluid concentrations of IL-8 and/or the presence of endometriosis. We conclude that IL-8 is an important factor that may contribute to the pathogenesis of endometriosis possibly by promoting neovascularization. This information can be a guide in the development of new therapeutic approaches for the treatment of endometriosis.      

Peritoneal fluid concentrations of interleukin-8 in patients with endometriosis depend on the severity of the disorder and are higher in the luteal phase

BACKGROUND: Previous evaluations of the relationship between the concentrations of interleukin-8 (IL-8) in the peritoneal fluid and endometriosis led to non-consistent results. Our purpose was to investigate the correlation of the concentrations of IL-8 in the peritoneal fluid with the stage of endometriosis, the presence of red lesions and the phase of the menstrual cycle. METHODS: Ninety-two patients with infertility (n = 87) or undergoing sterilization (n = 5) had peritoneal fluid samples collected at laparoscopy. IL-8 determinations were performed using an enzyme-linked immunosorbent assay. RESULTS: The concentrations of IL-8 in the peritoneal fluid of the 68 women with endometriosis were not significantly different from those of the 24 controls. Patients with moderate/severe stages had IL-8 significantly higher than controls (P = 0.008) and marginally higher than patients with minimal/mild endometriosis (P = 0.053). Concentrations of IL-8 were significantly higher in patients than in controls in the luteal phase. Red lesions were associated with significantly increased levels of peritoneal fluid IL-8 only in the luteal phase. CONCLUSIONS: Our findings reinforce the importance of IL-8 in the pathogenesis of endometriosis.     

Role of IL-18 in pathogenesis of endometriosis

BACKGROUND: Endometriosis is a complex disease associated with a wide range of immune responses, including pain, adhesion, exudation of peritoneal fluid, elevation of cytokine levels and generation of autoantibodies. Interleukin (IL)-18 is a strong pleiotropic cytokine known to be involved in various immune diseases. The aim of this study is to elucidate the role of IL-18 in the pathogenesis of endometriosis. METHODS: IL-18 and IL-1beta concentrations were measured in the peritoneal fluid and sera of 39 endometriosis patients and 15 control women. Expression of IL-18 and IL-18 receptor alpha-chain (IL-18Ralpha) was analysed in endometriotic tissues immunohistochemically. The effects of IL-18 on cyclooxygenase (COX)-II gene expression were analysed in peritoneal fluid monocytes and endometriotic cells of endometriosis patients. RESULTS: IL-18 concentrations in the peritoneal fluid of endometriosis patients averaged 592.57 +/- 108.27 pg/ml, significantly higher than 260.50 +/- 55.88 pg/ml in non-endometriotic samples. IL-18 concentrations in the serum did not differ significantly between endometriosis and control patients. Similarly, no significant differences were observed in IL-1beta concentrations in either the peritoneal fluid or the serum. IL-18 and IL-18Ralpha were expressed in endometriotic tissues. IL-18Ralpha expression was also observed in cells infiltrating into the inflammatory area of the endometriosis patients. COX-II was induced in peritoneal fluid monocytes and in endometriotic cells in response to IL-18 stimulation. CONCLUSIONS: The elevation of IL-18 in the peritoneal fluid of endometriosis patients and the induction of COX-II in peritoneal monocytes by IL-18 suggest that IL-18 plays a pathogenic role in endometriosis.     

The Peritoneal Fluid Levels of Interleukin-12 in Women with Endometriosis

PROBLEM: Interleukin-12 (IL-12) is produced mainly by monocytes/macrophages, and it induces proliferation and cytotoxicity of T-cells and natural killer cells. In women with endometriosis, natural killer cell activity in the peritoneal fluid is significantly decreased. We aimed to measure the peritoneal fluid level of IL-12 in endometriosis. METHOD OF STUDY: We measured IL-12 levels in peritoneal fluid samples from women with or without endometriosis and in supernatants from endometrial stromal, ovarian stromal, and mesothelial cell cultures, using a high-sensitivity enzyme-linked immunosorbent assay. RESULTS: The median concentration of IL-12 in the peritoneal fluid of women with endometriosis was 1.1 pg/ml (range, 0.2–5.5) and was 1.6 pg/ml (range, 0.4-2.8) in women without endometriosis, not a statistically significant difference. IL-12 was not detected in the supernatants of endometrial stromal, ovarian stromal, and mesothelial cell cultures. CONCLUSION: Concentrations of IL-12 in the peritoneal fluid of women with or without endometriosis are low, but they are detectable and are not affected significantly by the presence of endometriosis.     

Interferon-Gamma (IFN-γ) and Interleukin-6 (IL-6) in Peritoneal Fluid and Macrophage-Conditioned Media of Women With Endometriosis

PROBLEM: The presence of the various cytokines in human peritoneal fluid has been incompletely evaluated. Changes in cytokine levels may be related to activation of peritoneal macrophages, development of endometriosis, and infertility. This study assesses peritoneal fluid levels of interferon gamma (IFN-γ) and interleukin-6 (IL-6), and peritoneal macrophage production of IL-6, in women with and without endometriosis. METHOD: Peritoneal fluid was obtained from 62 women at the time of diagnostic or operative laparoscopic surgery for benign gynecologic disease. Peritoneal macrophages were isolated, cultured for 24 h, and the culture media collected. IFN-γ and IL-6 levels in peritoneal fluid samples and macrophage conditioned media were determined by commercial ELISA. RESULTS: IL-6 was significantly higher in the macrophage conditioned media of women with endometriosis as compared with controls. IL-6 levels were fourfold higher in early stage endometriosis (P < 0.05) and eightfold higher in advanced endometriosis. There were no significant differences between groups in the peritoneal fluid levels of IL-6 or IFN-γ. CONCLUSIONS: Peritoneal macrophage IL-6 secretion is increased in women with endometriosis, and appears to correlate with disease stage. IFN-γ does not appear to be responsible for the activation of macrophages in women with endometriosis.     

IL-1β TNF-α, and IL-2 in Peritoneal Fluid and Macrophage-Conditioned Media of Women With Endometriosis

PROBLEM : The presence of various cytokines in human peritoneal fluid has been incompletely evaluated. Changes in cytokine levels may be related to the development of endometriosis, infertility, and activation of peritoneal macrophages. This study assesses levels of IL-1β, IL-2, and TNF-α in peritoneal fluid and macrophage conditioned media of women with endometriosis. METHOD : Peritoneal fluid was collected from 51 women at the time of diagnostic or operative laparoscopy for benign gynecologic disease. Peritoneal macrophages were isolated, cultured for 24 h, and the culture media collected. IL-1β, IL-2, and TNF-α levels were determined by commercial ELISA kits. RESULTS : The mean concentration of IL-1β and TNF-α was significantly higher in macrophage conditioned media of patients with endometriosis (P < 0.02). However, there were no significant changes in peritoneal fluid cytokine levels. Peritoneal macrophage concentrations were also higher in patients with endometriosis. CONCLUSION : This study supports the concept that endometriosis is associated with activation of peritoneal macrophages, and a higher concentration of these cells. This activation is reflected by the increased levels of cytokines found in macrophage conditioned media. The absence of significant changes in peritoneal fluid cytokine levels would seem to indicate that the above derangements are not responsible for the development or progression of endometriosis.     

Peritoneal fluid cytokines and the relationship with endometriosis and pain

It is generally accepted that the current scoring system forendometriosis has little correlation with clinical symptomssuch as pain, and therefore we may deduce that either endometriosisdoes not cause pain, or that the current scoring system doesnot indicate the biological activity of the disease. Pain mayoccur because the presence of endometriosis produces an intraperitonealinflammatory response, and several studies have shown that thecytokine content of peritoneal fluid differs between women withand without endometriosis. We studied the relationship betweentumour necrosis factor a (TNFa), platelet-derived growth factor(PDGF), interleukin (IL)-6, IL-4 and TNF (a and P) activityin peritoneal fluid and the clinical history of pain and infertility.TNFa concentrations were increased in peritoneal fluid of womenwith endometriosis and of infertile women; PDGF concentrationswere increased in peritoneal fluid of parous women; EL-6 wasincreased in peritoneal fluid of women with adhesions; IL-4was absent from peritoneal fluid. PDGF and IL-6 concentrationswere cycle related, with the highest amounts in the menstrualand proliferative phases respectively. We failed to demonstrateany association between concentrations of cytokines in vitroand pain symptoms or severity of endometriosis.     

Peritoneal interleukin-10 increases with decrease in activated CD4+ T lymphocytes in women with endometriosis

This study was performed to determine whether peritoneal T cells are suppressed in the CD4+ or CD8+ T cell subpopulation and whether they are Th1 or Th2 predominant in women with endometriosis. Immune cells in the peritoneal fluid (PF) were obtained from women undergoing laparoscopy for endometriosis or tubal ligation. Three-colour flow cytometry was utilized for immunophenotyping of peritoneal fluid mononuclear cells (PFMC). Concentrations of interleukin (IL)-4, IL-5 and interferon-gamma (IFN-gamma) produced by PFMC with and without mitogen stimulation and concentrations of IL-10 and IL-12 were measured in PF. The peritoneal T lymphocytes were predominantly of the Th1 type that produced much more IFN-gamma but less IL-4 or IL-5 in women with or without endometriosis. The decrease in peritoneal lymphocytes was significant in the HLA-DR+ CD4+ CD3+ subpopulation and the concentrations of peritoneal IL-10 and IL-12 were significantly elevated in women with early stage endometriosis. There was impaired IL- 5 production by PFMC after phytohaemagglutinin stimulation in women with advanced stage endometriosis. We concluded that the activated peritoneal CD4+ Th1 cells from the women with endometriosis were decreased in number. The suppression of these T cells may be due to the elevation of IL-10 and IL-12 in the peritoneal fluid.      

Increased levels of interleukin-15 in the peritoneal fluid of women with endometriosis: inverse correlation with stage and depth of invasion

BACKGROUND: Interleukin (IL)-15 is a novel cytokine with immunoregulatory and angiogenic properties. We compared IL-15 levels in the peritoneal fluid (PF) of women with and without endometriosis. METHODS: PF samples were obtained from 55 women with endometriosis (23 with superficial peritoneal implants, 19 with deep endometriotic implants and 13 with ovarian endometriomas). Eighteen women with normal pelvic anatomy undergoing tubal sterilization served as controls. RESULTS: PF IL-15 concentrations were increased in women with endometriosis (2.7 +/- 0.5 pg/ml) versus controls (2.1 +/- 0.3 pg/ml; P < 0.001). However, IL-15 levels were higher in women with superficial peritoneal implants (2.9 +/- 0.5 pg/ml) than women with deep endometriotic implants (2.6 +/- 0.4 pg/ml; P = 0.01) or ovarian endometriomas (2.2 +/- 0.4 pg/ml; P < 0.001). IL-15 was also higher in women with deep implants than in those with endometriomas (P < 0.05). PF IL-15 correlated inversely with both depth of invasion (r = -0.52) and the stage of endometriosis (r = -0.42). PF IL-15 levels demonstrated little variation during the menstrual cycle, and did not discriminate between women with infertility or pelvic pain. CONCLUSION: PF IL-15 levels are increased in women with endometriosis. However, IL-15 levels are inversely correlated with the depth of invasion and disease stage, suggesting a possible role for this cytokine in the early pathogenesis of endometriosis.