妊娠早期蜕膜组织巨噬细胞分泌IL-10/IFN-γ功能的特征

目的:探讨正常妊娠早期母胎界面蜕膜组织与外周血中细胞因子分泌的格局以及单核/巨噬细胞分泌IL-10/IFN-γ的功能特征。方法:收集9例接受人工流产的正常早孕妇女外周血和蜕膜组织,用MACS法富集单核/巨噬细胞,用ELISPOT法检测细胞因子。结果:在正常妊娠早期,母胎界面蜕膜组织分泌IL-10的细胞细胞和分泌IFN-γ细胞比值显著高于外周血;蜕膜巨噬细胞分泌IL-10数量明显高于外周血单核细胞,亦明显高于蜕膜组织非巨噬细胞。结论:蜕膜组织分泌细胞因子格局的特征与外周血有差异,可能是母胎免疫耐受形成的重要途径。蜕膜巨噬细胞可通过增强IL-10的分泌调节母胎界面细胞因子格局,有利于诱导母胎免疫耐受。     

Cytokine microenvironments in human first trimester decidua are dependent on trophoblast cells

OBJECTIVE: To compare cytokine expression profiles of decidua basalis (containing trophoblast cells) and decidua parietalis (without trophoblast cells) for determination of microenvironments in human first trimester decidua. DESIGN: Retrospective study. SETTING: School of Medicine, RWTH University of Aachen, Aachen Germany, and Bourgognekliniek Maastricht, Maastricht, The Netherlands. PATIENT(S): Forty-six women who had undergone elective first-trimester termination of viable pregnancy at 5 to 12 weeks. MAIN OUTCOME MEASURE(S): Quantitative cytokine protein analysis in decidual tissues by enzyme-linked immunosorbent assay, qualitative cytokine messenger (m)RNA analysis in isolated decidual cell samples, and comparative mRNA and protein analysis in tissues of decidua basalis compared with decidua parietalis. RESULT(S): Interleukin-2, interferon-gamma (Th-1), interleukin-4 (Th-2), and interleukin-1beta proteins are expressed in the human first-trimester decidua. Interleukin-2, interferon-gamma, and interleukin-4 mRNA mainly derive from the decidual tissue leukocytes. Interleukin-1beta mRNA is expressed by all decidual cell types. Interferon-gamma mRNA and protein is detected predominantly in the decidua basalis, which contains trophoblast cells. CONCLUSION(S): Microenvironments are established topographically by different expression of cytokines in decidua basalis and decidua parietalis. These locally specific patterns are indicative of fetomaternal cross-talk. Higher interferon-gamma concentrations in decidua basalis may influence leukocyte differentiation (e.g., macrophage activation) and trophoblast invasion (e.g., by induction of expression of major histocompatibility complex).     

正常早孕蜕膜非免疫细胞免疫活性的实验研究

目的 了解蜕膜中非免疫细胞的免疫活性。方法 取蜕膜中非免疫细胞培养的上清液,用放射免疫方法（ＲＩＡ）测定其对人外周血Ｔ细胞的转化、自然杀伤（ＮＫ）细胞的杀伤率、淋巴细胞分泌白细胞介素２（ＩＬ－２）及Ｂ细胞分泌ＩｇＢ的影响。结果 蜕膜非免疫细胞培养的上清液可使Ｔ细胞、ＮＫ细胞、Ｂ细胞的免疫学活性不同程度地受到抑制,其最大抑制率分别为２２．７％、５２．３％和１４．８％,但对淋巴细胞ＩＬ－２的分泌无显著     

Expression profiling of vitamin D receptor in placenta, decidua and ovary of pregnant mice

Objectives

The presence of vitamin D receptor (VDR) and the identification of localized vitamin D3 synthesis in placenta and decidua implicate the importance of vitamin D3 in reproductive function. There is, however, no data on the expression profile of VDR in the mouse placenta and endometrium throughout the pregnancy period.

Study design

In the present work expression of VDR in reproductive tissues of pregnant mice at different gestational phases has been addressed. Expression of VDR was determined by semi-quantitative RT-PCR, Western blotting and immunohistochemistry.

Results

The results showed that VDR mRNA and protein were expressed in decidua, placenta and ovary throughout the pregnancy. VDR gene expression in placenta was significantly elevated in late pregnancy when compared to that of mid pregnancy. Additionally, VDR expression level in decidua rose significantly as pregnancy progressed from early to mid stages. VDR expression in decidua of pregnant mice was higher in comparison to endometrium of non-pregnant mice. Immunohistochemical analysis revealed that VDR protein is consistently expressed by luminal and glandular epithelial cells of decidua, giant cells, glycogen rich cells and labyrinth cells of placenta and by almost all follicular cell types of ovary. Surveying the expression of VDR at the protein level by Western blotting confirmed PCR results.

Conclusion

It seems that expression of VDR in reproductive organs is finely tuned during pregnancy indicating its eminent role in reproductive biology.     

Expectant management of incomplete abortion in the first trimester

Objective

To evaluate the effectiveness and acceptability of expectant management of induced and spontaneous first trimester incomplete abortion.

Methods

A prospective observational trial, conducted between June 2006 and November 2007, of 2 groups of patients diagnosed with an incomplete abortion: 66 patients who had received misoprostol for an induced abortion (group 1) and 30 patients who had had a spontaneous abortion (group 2). Transvaginal ultrasound was performed weekly. The success rate (complete abortion without surgery), time to resolution, duration of bleeding and pelvic pain, rate of infection, number of unscheduled hospital visits, and level of satisfaction with expectant management were recorded.

Results

The incidence of complete abortion was 86.4% and 82.1% in groups 1 and 2 respectively at day 14 after diagnosis, and 100% in both groups at day 30 (two group 2 patients underwent curettage and were excluded from the analysis). Both groups reported 100% satisfaction with expectant management, although over 90% of the women reported feeling anxious.

Conclusion

Expectant management for incomplete abortion in the first trimester after use of misoprostol or after spontaneous abortion may be practical and feasible, although it may increase anxiety associated with the impending abortion.     

The production of prostanoids and the identification of macrophages in human decidua vera tissue

Summary In decidual tissues of first trimester therapeutic abortions we have identified a large macrophage population that is stained by a specific antibody (Ki-M8). Also, we evaluated the capacity of decidual cells, obtained before and after labor, to synthesize prostanoids from radiolabeled arachidonic acid. Decidua collected after vaginal delivery produced significantly more prostaglandin (PG) F_2α , PGE₂ and thromboxane (TX) B₂ (7.4 ± 1.3, 7.2 ± 1.3, 11.7 ± 1.1 pmol/mg protein/4h, mean ± SD, respectively) than did tissues from women who had caesarean sections (3.3 ± 0.8, 1.4 ± 0.5, 6.3 ± 1.8 pmol/mg protein/4h, respectively). Moreover, PGF_2α and PGE₂ were rapidly converted to their 13,14-dihydro-15-ketometabolites particularly in tissues obtained after vaginal delivery. All this lends further support to the proposition that decidual activation may be important in the onset of labor.     

Apoptosis of extravillous trophoblast cells limits the trophoblast invasion in uterine but not in tubal pregnancy during first trimester

During the first trimester of pregnancy extravillous trophoblast cells (EVT) invade the maternal decidua. Invasion normally is reduced from the second trimester onwards and stops in the inner third of the myometrium. By contrast, in extrauterine tubal pregnancy, trophoblast invasion may even penetrate the tubal wall, which ultimately leads to the rupture of the fallopian tube. Induction of apoptosis of EVT cells, by maternal immune competent cells, may be an important mechanism to limit EVT invasion in uterine pregnancy. Tissue specimens from first and second trimester uterine pregnancy and first trimester tubal pregnancy were analyzed for apoptosis by TUNEL- and M30-staining. By immunohistochemical double labelling, maternal leukocyte subtypes were co-localized to apoptotic cells and in this context, the number of CD56(+)NK cells was analyzed. Our data show that apoptosis is confined to the decidua basalis. Most apoptotic cells are single cytokeratin-positive epithelial cells residing in the stromal compartment. Consequently these cells can only be EVT cells. Maternal leukocytes are not apoptotic. They are located in close contact to apoptotic cells. The number of apoptotic cells in the second trimester (1.8+/-0.7 per cent) is reduced compared to first trimester (5.6+/-0.7 per cent) of uterine pregnancy. In parallel, the number of NK cells declines from first (24.4+/-2.9) to second (12.4+/-1.8) trimester. Furthermore, apoptosis is significantly reduced in ectopic (0.9+/-0.3 per cent) compared to eutopic first trimester pregnancies. Consequently, we suggest that in first trimester uterine pregnancy, induction of EVT cell apoptosis by the maternal immune system is one mechanism to limit EVT invasion. During the second trimester, in parallel to declining numbers of NK cells, the mechanism changes. However, in tubal pregnancy due to differing immunological microenvironments at the ectopic implantation site, apoptosis induction fails, which deleteriously may result in uncontrolled invasion and penetration of the tubal wall.     

早孕蜕膜及绒毛组织中趋化因子CXC受体3、4及其配体的表达变化和意义

目的 探讨早孕蜕膜及绒毛组织中趋化因子CXC受体(CXCR)3、4及其配体CXCL9、CXCL10和CXCL12的表达变化和意义.方法 体外分离正常早孕蜕膜组织中单个核细胞,免疫磁珠分选试剂盒纯化CD+56自然杀伤(NK)细胞,流式细胞仪分析其纯度和表型;RT-PCR技术榆测早孕蜕膜NK细胞中CXCR3和CXCR4、早孕蜕膜及绒毛组织中CXCL9、CXCL10、CXCL12的表达情况;免疫组化链霉菌抗生物素蛋白-过氧化物酶连接(SP)法检测正常子宫内膜、早孕蜕膜组织中CXCL9和CXCL10的表达及CD+56NK细胞的分布情况,分析CXCL9、CXCL10表达量(灰度值)与CD+56NK细胞数的相关性.结果 分离纯化的早孕蜕膜NK细胞中,98.7%的细胞表型为CD56bright;早孕蜕膜NK细胞中有CXCR3和CXCR4表达;早孕蜕膜组织中有CXCL9、CXCL10表达,早孕绒毛组织中有CXCL12的表达.分泌期子宫内膜中CXC19、CXCL10表达最为56±43、59±47,较增生期子宫内膜的16±18、8±14明显升高,差异有统计学意义(P<0.05),而早孕蜕膜组织中CXCL9、CXCL10表达量为143±35、158±29,较分泌期子宫内膜进一步升高(P<0.05);分泌期子宫内膜中NK细胞数量为(60±20)个,增生期子宫内膜中NK细胞数量为(23±4)个,两者比较,差异有统计学意义(P<0.05),早孕蜕膜中NK细胞数量为(114 ±15)个,较分泌期子宫内膜进一步增多(P<0.05);子宫内膜和蜕膜组织中CXCL9、CXCL10表达量与组织中CD56+细胞数呈正相关关系(rcxL9=0.88,P<0.05;rcxcL10=0.86,P<0.05).结论 早孕蜕膜及绒毛组织中表达的CXCL9、CXCL10及CXCL12可能通过与CD56+NK细胞表面对应的趋化因子受体CXCR3、CXCR4结合而影响早孕期母-胎界面中CD56+NK细胞的聚集,从而对母-胎间免疫平衡起调控作用.     

Placental blood flow mapping in the first trimester of human pregnancy

Human placental circulation in the first trimester of pregnancy remains a subject of scientific debate. Most of the acquired knowledge on the physiological background of maternal-fetal interface relies on old studies, most of them performed in non-hemochorial placentas and using light and electron microscopy for the pathological specimens' evaluation. Recently, some ultrasonographic research in this field, using pulsed-wave and/or colour Doppler, yielded contradictory results. The availability of these non-invasive methods should allow the in vivo investigation of placental circulation since early phases of human pregnancy. A more recent technique - Power Doppler - should be preferred considering its resolution, angle-independence, higher sensitivity for lower velocities and absence of known harmful bioeffects in the early periods of gestation. We provide an overview on first trimester maternal-fetal circulation, assessed by non-invasive techniques, namely by transvaginal power Doppler ultrasonography, trying to contribute to a better understanding of the anatomo-physiological aspects of human placental circulation.     

外源性细胞因子对早孕蜕膜T细胞免疫活性的研究

目的 :探讨外源性细胞因子对早孕蜕膜T细胞免疫活性作用的方式。方法 :在培养的蜕膜细胞中加入γ干扰素、白细胞介素 2、白细胞介素 6和表皮生长因子 ,共同培养 12、2 4、4 8h后收集上清液 ,并加入培养的T细胞中 ,用氚 胸苷 (3 H TdR)掺入法测定T细胞的转化值。结果 :在蜕膜与细胞因子共同培养的上清液作用下 ,各实验组T细胞的转化有不同程度的升高 ,有的T细胞转化效应显著增强 (P <0 .0 5 ) ,但增强的T细胞转化效应未显示出与作用于蜕膜的细胞因子种类、浓度及作用时间有相关性。结论 :在外源性细胞因子的作用下 ,早孕蜕膜细胞可以增强T细胞的转化作用 ,从而破坏蜕膜免疫微环境细胞因子网络的平衡状态 ,危害妊娠。     

Yolk sac size and embryonic heart rate as prognostic factors of first trimester pregnancy outcome

OBJECTIVES: To investigate embryonic heart rate (EHR) and yolk sac diameter (YSD) during the first trimester and their role as prognostic markers of first trimester pregnancy outcome. STUDY DESIGN: Prospective cohort study involving 219 women conducted in the 4th Academic Department of Obstetrics and Gynaecology, Aristotle University of Thessaloniki, Greece. Gestational age (GA) was determined ultrasonographically based on gestational sac diameter and crown-rump length. EHR and YSD were evaluated during the first 12 weeks and subsequently compared between the pregnancies that continued beyond the first trimester and those that resulted in spontaneous abortion. Receiver-operating characteristic (ROC) curves were used for the evaluation of the prognostic value of the combination of gestational age with embryonic heart rate and yolk sac diameter. RESULTS: The EHR and YSD were significantly correlated to advancing gestational age (p<0.001) in pregnancies continuing beyond 12 weeks. Pregnancies that resulted in spontaneous abortion exhibited a statistically significant lower EHR (p<0.001), smaller YSD (p=0.001) or absent yolk sac. ROC curve analysis demonstrated the predictive value of the combination of GA with EHR (area under the ROC curve: 0.971, p<0.001) and GA with YSD (area under the ROC curve: 0.858, p<0.001) for first trimester pregnancy outcome. CONCLUSIONS: EHR and YSD progressively increase in healthy pregnancies during the first trimester. Embryonic bradycardia and absence of yolk sac or even a smaller yolk sac diameter than expected for any gestational age are predictors of poor pregnancy outcome during the first 12 weeks.     

妊娠早期孕妇血清解整合素-金属蛋白酶12水平变化与妊娠结局

值为0.29,明显低于正常单胎孕妇,差异有统计学意义(P<0.05).结论 早孕期孕妇血清中ADAM 12-S水平随孕周增加而上升,可用于非整倍体常染色体异常的筛查,同时还可预测胎停育、异位妊娠等早期妊娠丢失;ADAM 12-S可作为产前筛查指标,建议在孕9～12周时进行.     

Effect of maternal age on spontaneous abortion during the first trimester in Northeast China

Objective: We aimed to determine the relationship between fetal chromosomal abnormalities and maternal age among spontaneous first trimester abortions in women in Northeast China.

Methods: We evaluated 497 chorionic villi samples from patients with a history of spontaneous abortion during the first trimester. We divided the samples into five groups according to the maternal age: <25, 25–29, 30–34, 35–39, and ≥40 years. We identified chromosomal abnormalities by fluorescence in situ hybridization.

Results: Among the 497 chorionic samples of spontaneous abortion, 180 (36.22%) had fetal chromosomal abnormalities. Patients aged ≥40 years had a significantly higher percentage (60.61%) of fetal chromosomal abnormalities compared with the other groups. More women in the ≥40 and 35- to 39-year groups had a history of three consecutive miscarriages and 10 kinds of abnormalities. The most frequent aneuploidy was trisomy 22, followed by trisomy 16.

Conclusions: These results revealed that the kinds of fetal abnormalities, numbers of abortions, and chromosomal abnormality rates increased with increasing maternal age. The most common trisomy types in spontaneous abortions were closely related to maternal age. We hypothesize whether the larger probability of chromosomal abnormalities is due to increased mutation rate with maternal age, or due to a worse in-utero conditions.     

Th17及相关细胞因子在妊娠早期母-胎免疫耐受中的作用

目的:研究Th17细胞及相关细胞因子IL-22、IL-23在人孕早期不明原因的自发性流产母-胎免疫耐受中的作用。方法:以10例正常人工流产患者为正常早孕组,收集18例染色体正常自发性流产患者为自然流产组。分离两组患者外周血单个核细胞及蜕膜组织免疫细胞,用流式细胞术检测上述细胞中Th17细胞比例及相关细胞因子的表达,探讨其与自发性流产的关系。结果:自然流产组外周血Th17细胞比例显著高于正常早孕组(P<0.05),IL-23显著低于正常早孕组(P<0.01),但两组IL-22无显著差异;自然流产组蜕膜局部Th17细胞比例显著高于正常早孕组(P<0.01),IL-23亦显著高于正常早孕组(P<0.01),IL-22则显著低于正常早孕组(P<0.01)。结论:Th17细胞在孕早期蜕膜局部及外周血中的数量增多可能导致母胎免疫耐受失衡,在早期自然流产中发挥重要的正诱导作用,而IL-22、IL-23等细胞因子在其中的调控效应则具有明显的微环境相关性。     

白血病抑制因子在早期妊娠、先兆流产及难免流产患者蜕膜组织中表达的研究

目的探讨白血病抑制因子(LIF)在蜕膜组织中的表达,及其与早期妊娠、流产的关系. 方法采用放射免疫方法检测正常早孕妇女(正常早孕组)、先兆流产妇女(先兆流产组)及难免流产妇女(难免流产组)的血清孕酮及人绒毛膜促性腺激素(hCG)水平,并采用逆转录-聚合酶链反应(RT-PCR)技术对3组孕妇蜕膜组织中LIF-mRNA的表达进行定量分析.结果 (1)孕酮及hCG水平在3组孕妇间的比较正常早孕组孕妇血清中孕酮、hCG水平分别为(91.5±27.2) nmol/L、(69.9±14.9) kU/L,先兆流产组孕妇分别为(88.4±24.7) nmol/L、(57.6±11.2) kU/L,两组孕妇血清孕酮、hCG水平比较,差异均无显著性(P>0.05);而难免流产组孕妇血清孕酮、hCG水平分别为(33.1±19.6) nmol/L、(10.3±3.2) kU/L,与前两组分别两两比较,差异均有极显著性(P<0.01).(2)LIF-mRNA平均相对含量在3组孕妇间的比较正常早孕组孕妇为2.10±0.32;先兆流产组孕妇为1.92±0.20;难免流产组孕妇为0.70±0.06.正常早孕组与先兆流产组比较,差异无显著性(P>0.05).而难免流产组与前两组分别行两两比较,差异均有显著性(P<0.05).结论 LIF-mRNA在早期妊娠蜕膜组织中的表达量降低,可能是导致hCG及孕酮分泌下降,最终造成难免流产的原因之一.     

早期自然流产患者绒毛KiSS-1、MMP-9和MMP-2基因及其蛋白表达

目的研究早期自然流产患者绒毛肿瘤转移抑制基因KiSS-1与基质金属蛋白酶(MMP-9、MMP-2)mRNA及蛋白的表达,探讨其与自然流产的关系。方法采用半定量RT-PCR、免疫印迹法检测30例正常早期妊娠和30例早期自然流产患者绒毛KiSS-1、MMP-9和MMP-2的mRNA及蛋白水平,用明胶酶谱分析法检测上述样本孵育液MMP-9、MMP-2的酶活性。结果(1)自然流产患者绒毛组织中MMP-9及MMP-2mRNA表达水平低于正常早期妊娠组(P<0.05),KiSS-1mRNA的表达高于正常早期妊娠组差异有统计学意义(P<0.05);(2)Westernblotting分析显示自然流产患者绒毛MMP-9、MMP-2蛋白表达低于正常早期妊娠差异有统计学意义(P<0.05),KiSS-1蛋白表达高于正常早期妊娠差异有统计学意义(P<0.05);(3)明胶酶谱分析结果显示,自然流产患者绒毛MMP-9、MMP-2酶活性低于正常早期妊娠者差异有统计学意义(P<0.05)。结论自然流产患者绒毛促浸润基因MMP-9、MMP-2mRNA及其蛋白表达水平降低,同时抑制浸润基因KiSS-1mRNA及其蛋白表达增加,造成滋养细胞浸润能力下降,可能与自然流产的发生有关。