The distribution and possible function of gamma interferon-immunoreactive cells in normal endometrium and myometrium

Summary T-lymphocytes are present in normal endometrium, where they may have a role in the control of glandular maturation. T-cell activity could be related to the local secretion of cytokines such as gamma interferon, which has an anti-proliferative effect on endometrial epithelial cells in vitro. We have examined gamma interferon immunoreactivity and T-cell distribution in 24 normal pre-menopausal uteri. Endometrial appearances were representative of all stages of the menstrual cycle. Most cells in the lymphoid aggregates in the stratum basalis were stained by T-cell and gamma interferon antisera. T-lymphocytes were also scattered in glandular epithelium and throughout the stroma of basal and functional layers; immunoreactivity for gamma interferon was less consistent in these cells. There was no alteration in the intensity or distribution of gamma interferon staining in different phases of the menstrual cycle. Endometrial granulocytes (K-cells) present mainly in the late secretory endometria were not reactive with the gamma interferon antiserum. In addition to endometrial staining, T-cells were distributed in all areas of the myometrium in most uteri, and many myometrial lymphocytes were gamma interferon positive. These results support a role for gamma interferon in endometrial physiology, possibly as an inhibitor of epithelial proliferation.     

BCL—2和增殖细胞核抗原子在子宫肌瘤中的表达

目的：研究bcl-2蛋白和增殖细胞核抗原（PCNA）在子宫肌瘤和正常子宫肌层中的表达及其在肌瘤发生发展中的作用。方法：用免疫组织化学ABC法检测40例子宫肌瘤组织和正常子宫平滑肌组织中的bcl-2蛋白和PCNA的表达。结果：bcl-2和PCNA在肌瘤组织中表达均明显强于正常 子宫平滑肌组织（P＜0．001）。结论：bcl-2和PCNA在肌瘤中过度表达以及两者的共同作用,可能在子宫肌瘤的发生发展过程中起重要作用。     

Oxytocin mobilizes calcium from a unique heparin-sensitive and thapsigargin-sensitive store in single myometrial cells from pregnant rats

Intracellular free Ca²⁺ concentration ([Ca²⁺]_i) was monitored using the fluorescence from the dye Fura-2-AM in single myometrial cells from pregnant rats. Oxytocin and acetylcholine applied to the cell evoked an initial peak in [Ca²⁺]_i followed by a smaller sustained rise which was rapidly terminated upon removal of acetylcholine or persisted after oxytocin removal. A Ca²⁺ channel blocker (oxodipine) and external Ca²⁺ removal decreased both the transient and sustained rises in [Ca²⁺]_i suggesting that Ca²⁺ influx through L-type Ca²⁺ channels participated in the global Ca²⁺ response induced by oxytocin. However, the initial peak in [Ca²⁺]_i produced by oxytocin was mainly due to Ca²⁺ store release: it was abolished by inclusion of heparin [which blocks inositol 1,4,5-trisphosphate (InsP ₃) receptors] in the pipette (whole-cell recording mode of patch-clamp) and external application of thapsigargin (which blocks sarcoplasmic reticulum Ca²⁺-ATPases). In contrast, the transient Ca²⁺ response induced by oxytocin was unaffected by ryanodine. Moreover, caffeine failed to induce a rise in [Ca²⁺]_i but reduced the oxytocin-induced transient Ca²⁺ response. The later sustained rise in [Ca²⁺]_i produced by oxytocin was due to the entry of Ca²⁺ into the cell as it was suppressed in external Ca²⁺-free solution. The Ca²⁺ entry pathway is permeable to Mn²⁺ ions, in contrast to that described in various vascular and visceral smooth muscle cells. Oxytocin-induced Ca²⁺ release is blocked by the oxytocin antagonist d(CH₂)₅[Tyr(Me)²,Thr⁴,Tyr-NH ₂ ⁹ ]OVT. The prolonged increase in [Ca²⁺]_i after oxytocin removal is rapidly terminated by addition of the oxytocin antagonist suggesting that oxytocin dissociation from its receptor is very slow. The oxytocin stimulation of [Ca²⁺]_i was insensitive to incubation with pertussis toxin, and blocked by a pipette solution containing anti-q/₁₁ antibody. These data show that myometrial cells possess an unique heparin-sensitive and thapsigargin-sensitive store that can be mobilized by activation of oxytocin receptors which couples with a Gq/G₁₁-protein to activate phospholipase C.     

Interaction between sex hormones and WNT/β-catenin signal transduction in endometrial physiology and disease

Wnt/β-catenin signalling plays a rate-limiting role in early development of many different organs in a broad spectrum of organisms. In the developing Müllerian duct, Wnt/β-catenin signalling is important for initiation, outgrowth, patterning and differentiation into vagina, cervix, uterus and oviducts. In adult life, sex hormones modulate Wnt/β-catenin signalling in the endometrium to maintain the monthly balance between estrogen-induced proliferation and progesterone-induced differentiation, and enhanced Wnt/β-catenin signalling seems to be involved in endometrial carcinogenesis. However, early in pregnancy enhanced Wnt/β-catenin signalling is prerequisite for proper implantation and invasion of trophoblast cells into endometrium and myometrium thus helping to form a placenta. Overall, it seems that tight control of Wnt/β-catenin signalling in time and space is important for initiation, development and normal function of the female reproductive tract. However, if Wnt/β-catenin signalling is not kept in check, it easily seems to initiate or contribute to development of a number of uterine disorders.     

不同痛经表现的子宫腺肌症患者外周血肿瘤标记物表达水平差异性分析

目的 分析不同痛经表现的子宫腺肌症患者外周血肿瘤标记物表达水平,探讨肿瘤标记物检测的临床意义.方法 回顾分析我院2010年3月-2012年9月收治的85例子宫腺肌症患者的临床病例资料,按痛经程度及痛经持续时间分组,检测不同痛经程度及不同痛经持续时间患者的糖类抗原125（CA125）、糖类抗原199（CA199）、癌胚抗原（CEA）水平.结果 痛经程度重、痛经程度轻、无痛经的CA125、CA199、CEA水平组间比较,差异有统计学意义（P＜0.05）,CA125、CA199、CEA的高低排次为痛经程度重＞痛经程度轻＞无痛经;痛经持续时间长、痛经持续时间短、无痛经的CA125、CA199、CEA水平组间比较,差异有统计学意义（P＜0.05）,CA125、CA199、CEA的高低排次为痛经持续时间长＞痛经持续时间短＞无痛经.结论 子宫腺肌症患者有无痛经、痛经程度、痛经持续时间与外周血肿瘤标记物表达水平密切相关,连续、动态和联合检测肿瘤标记物水平具有重要的临床意义.     

临产后子宫体部平滑肌高表达HSP27和αB-crystallin

 ［摘要］目的:研究临产前后子宫体部平滑肌热休克蛋白27（HSP27）和α-晶体蛋白B链（αB-crystallin）的表达变化,探讨其在临产后子宫平滑肌收缩中的可能作用。方法:采用比较蛋白质组学技术鉴定出足月未临产和自然临产人子宫体部平滑肌中HSP27和αB-crystallin蛋白存在差异性表达,应用RT-PCR、免疫组化和免疫印迹方法对临产前后子宫体部平滑肌中HSP27蛋白的表达进行验证。结果:临产后子宫体部平滑肌高表达HSP27和αB-crystallin（P<0.05）。足月未临产和临产人子宫体部平滑肌双向凝胶电泳图谱均存在4个分子量相等、等电点不同的HSP27蛋白斑点,ImageMaster 2D Platinum 软件分析仅有1个HSP27蛋白斑点具有显著差异（P<0.05）,余3个HSP27蛋白斑点无显著差异（P>0.05）。RT-PCR、免疫组化和免疫印迹对HSP27蛋白验证结果与蛋白质组学一致。结论: HSP27和αB-crystallin蛋白在临产后人子宫体部平滑肌高表达,提示这2个小分子热休克蛋白可能参与了临产后子宫体部平滑肌细胞的收缩事件。     

A Quantitative Evaluation of Active Gelatinolytic Sites in Uterine Endometrioid Adenocarcinoma Using Film in situ Zymography: Association of Stronger Gelatinolysis with Myometrial Invasion

Collagen matrix degradation by malignant tumor cells plays an essential role in the process of tumor invasion and metastasis. The purpose of this study was to detect in situ gelatinase activity in endometrioid adenocarcinomas of the uterine corpus. In order to carry out quantitative evaluation, autoexposure tune (AET) on gelatin-coated film (film in situ zymography: FIZ) was measured. The gelatinase activity was located primarily within cancers and was prominently suppressed by the addition of a chelating agent to the film. This suggests that matrix metalloproteinases (MMPs) play an important role in the gelatinase activity. The gelatinase activity in the normal endometrium is almost negligible, despite positive immunoreactivity for MMP-2 and -9. Tumor tissues that had invaded more than half of the myometrium showed significantly higher activity than those that had invaded less than half. There was no significant difference in gelatinase activity among tumor stages, grades, vessel invasion or immunoreactivity for MMPs, with the exception that stage 2b cancers showed higher activity than stage la. The study suggested that the level of MMP-mediated gelatinolysis is an important factor for myometrial invasion in uterine endometrioid adenocarci- noma. Thus, a quantitative assessment of active gelatinolysis using FIZ and AET should be a useful tool in evaluating in situ matriolytic activity in local myometrial invasion by uterine endometrioid adenocarcinoma.     

Balance of amino acids in the pregnant human uterus at term

The nutritive and metabolic state of the human uterus at term is evaluated by measuring the uptake and release of amino acids. The subjects are ten healthy women with normal pregnancy undergoing elective caesarean section at term, before onset of labour. Free amino acids in arterial (radial artery) and venous (plexus of the uterine and ovarian veins) blood are determined and arteriovenous (AV) differences in each amino acid across the uterus are calculated. Generally the AV differences are negative, i.e. uterus at term releases amino acids in most cases. The human pregnant uterus at term is characterized by a release of amino acids rather than uptake. This indicates that they are in excess and are not needed in anabolic processes or as a fuel, even when the uterine tissue at term is supposed to be preparing for its grand performance, i.e. the delivery.     

Shortening increases spontaneous contractility in myometrium from pregnant women at term

OBJECTIVE: The purpose of this study was to determine whether shortening alters spontaneous contractility in myometrial strips that are obtained from pregnant women. STUDY DESIGN: Isometric contractions were measured in myometrial strips that were obtained at cesarean delivery from 14 pregnant women at term. After 2 hours of stretching, the strip lengths were decreased by 4%, 6%, or 8%. Spontaneous contractility was measured for 120 minutes with or without prostaglandin synthase inhibitor indomethacin (10 -5 mol/L), and the cumulative concentration response to oxytocin was determined. RESULTS: Contractility was increased by 29% and 34% in strips that shortened by 4% and 6%, respectively. Preincubation with indomethacin increased contractility by 15% in stretched strips and decreased contractility by 30% and 19% in 4% and 6% strips, respectively. Contraction frequency was increased by 26% and 53% for the strips that were shortened to 6% and 8%, respectively. These increases were prevented by indomethacin. The oxytocin responses were similar at all lengths. CONCLUSION: Shortening of myometrial strips from pregnant women at term increases spontaneous contractility by a mechanism that apparently involves prostaglandins.     

The presence and function of phosphodiesterase type 5 in the rat myometrium

OBJECTIVE: Cyclic nucleotide phosphodiesterases (PDEs) are a diverse enzyme group with multiple regulatory properties and wide tissue distribution. Such activity includes cyclic adenosine (cAMP) and guanosine monophosphate (cGMP) breakdown. The type 5 isoform (PDE-5, cGMP specific) is the target of specific antagonists (ie, sildenafil, Viagra). We tested the hypothesis that PDE-5 is present in rat myometrium and modulates myometrial activity. STUDY DESIGN: Full-thickness uterine wall was collected from nonpregnant (n=3) and pregnant Sprague-Dawley rats on days 10 (n=4), 17 (n=6), 22 nonlabor (n=5), and 22 during term labor (TL, n=4). Preterm labor (PTL, n=3) was induced in some animals on day 16 with 15 mg/kg mifepristone (RU 486). Tissue samples were prepared for Western blotting using a monoclonal antibody against rodent PDE-5. In a second series, cumulative doses of sildenafil (0.005, 0.05, 0.5, 5 mg/kg, intraperitoneal) were administered and the effect on uterine contractility recorded in vivo during term (TL, n=7) and preterm labor (PTL, n=6). Saline solution-injected rats provided temporal control. Uterine contractility was estimated from intrauterine pressure (IP) measured electronically with a sensor tip pressure catheter. Heart rate was recorded simultaneously using electrodes attached to the chest and connected to the same data acquisition system. RESULTS: PDE-5 immunoreactivity was present in the nonpregnant rat uterus and at all gestational times studied, although the expression was unaffected by either pregnancy or the state of labor (preterm or term). A dominant antibody-specific band was identified at 86 kd in the uterine samples, contrasting with lung where the 100-kd PDE-5 isoform was most abundant. Two additional lower molecular weight (55 and 32 kd) bands were also identified as antibody specific. Despite the lack of change in PDE-5 during pregnancy, sildenafil reduced IP during TL and PTL beginning at 0.5 mg/kg. The highest dose of sildenafil reduced IP during both TL and PTL by 45% and 59% of baseline, respectively (two-way analysis of variance, P<.01). This effect was not accompanied by changes in heart rate. CONCLUSION: PDE-5 is constitutively present in the rat uterine wall. There was no observed change in the PDE-5 protein expression throughout pregnancy. In contrast to the lung, the uterus expresses an 80-kd PDE-5 isoform. Sildenafil in pharmacologic doses inhibits mechanical uterine activity and might be of benefit if selectively used for treatment of preterm labor.