稽留流产与相关基因蛋白表达的关系研究

目的比较正常早孕组和稽留流产组两类妊娠女性的胎盘绒毛、蜕膜组织中Bcl-2、Bax基因蛋白表达情况,分析Bcl-2、Bax基因蛋白表达情况及其二者比值与胎盘绒毛、蜕膜组织中细胞凋亡的作用关系,为探索预防稽留流产发生相关的有效干预措施参考。方法搜集整理近年在北京航天总医院就诊的正常早孕和稽留流产女性93例,利用免疫组化学染色方法检测胎膜绒毛、蜕膜组织中Bcl-2、Bax基因蛋白表达情况,并定量分析其统计量、秩和检验以及相关性三类统计结果。结果无论是绒毛还是蜕膜组织,稽留流产组的Bax基因蛋白表达情况都要高于正常早孕组(0.21±0.37和0.1±0,0.5±0.79和0.14±0.18);绒毛组织两组的Bcl-2基因蛋白表达情况相近(2.95±0.31和2.94±0.42),但是稽留流产Bcl-2和Bax的比值要低于正常早孕组(26.92±8.55和29.41±4.2);蜕膜组织,稽留流产组的Bcl-2基因蛋白表达情况以及Bcl-2和Bax的比值均高于正常早孕组(2.48±0.9和1.47±1.02,18.66±13.09和14±10.22)。结论孕妇稽留流产原因与细胞凋亡相关基因高表达有关。     

Galectin-3基因在人孕早期绒毛和蜕膜中的表达及与妊娠的关系

目的 研究人早孕绒毛和子宫蜕膜中galectin-3基因mRNA和蛋白的表达，探讨其与滋养细胞浸润的关系。方法 采用半定量RT—PCR和Western-blot分别对70例（分为7个孕周组，每组10例）正常早孕绒毛和蜕膜中gal-3mRNA和蛋白表达水平进行检测 结果 在整个孕早期绒毛和子宫蜕膜中均有gal-3基因表达，且在绒毛和蜕膜中随着孕周的增加其表达总体呈上升趋势，两者表达趋势一致。结论 gal-3基因在滋养细胞和蜕膜表达中随孕龄增长而呈上调趋势，提示该基因在调控早孕滋养细胞的浸润行为中起重要的作用。     

人流组织和药流后刮出组织中hCG,hPL和PSβ1G免疫细胞化学检测

绒毛合体滋养层细胞分泌ｈＣＧ、ｈＰＬ、ＰＳβ１Ｇ，蜕膜中散在的绒毛外滋养层细胞中后二者含量很丰富＾〔１〕。它们可促进胎盘生长和提供胚胎营养。米非司酮抗早孕后，部分患者子宫持续出血，原因为蜕膜和绒毛残留＾〔２，３〕。本文研究药流刮出组织中绒毛合体滋养层和蜕膜中绒毛外滋养层细胞的功能，探讨它们与蜕膜残留和子宫出血的关系。     

自然流产中的早孕期蜕膜细胞Bcl-2 /Bax 比例异常

探讨早孕期蜕膜细胞细胞凋亡异常与妊娠失败的关系。应用TUNEL法测定凋亡的发生及凋亡细胞的定位，免疫组化法检测Bcl-2、Bax蛋白的表达和相互关系。结果发现：（1）正常早孕40d蜕膜组织细胞大量凋亡，Bcl-2蛋白表达量较低，Bax蛋白有较强表达。（2）正常早孕50d，凋亡细胞明显减少，Bcl-2的表达显著增强，Bax蛋白表达减弱。（3）早孕50d自然流产组，蜕膜组织大量凋亡，与同时期正常蜕膜组织相比，P＜0．01。Bcl-2蛋白表达明显降低，Bax蛋白表达明显增强，Bcl-2/Bax比例降低。早孕期蜕膜组织凋亡异常可能是自然流产的机制之一，Bcl-2/Bax途径可能是诱导早孕期蜕膜细胞凋亡的重要因素。     

白细胞介素-6在反复自然流产患者绒毛和蜕膜组织中的表达

目的:研究白细胞介素-6(IL-6)在反复自然流产患者绒毛与蜕膜组织中的表达情况,为防治反复自然流产提供理论依据.方法:应用免疫组织化学显色法和图像分析技术,对IL-6蛋白在反复自然流产患者(病例组)和正常人工流产者(对照组)绒毛和蜕膜组织中的表达进行定位和半定量分析;H-E染色法观察反复自然流产患者绒毛和蜕膜组织的形态学变化.结果:免疫组织化学显色显示,在绒毛组织中,IL-6蛋白定位于滋养层细胞的细胞质内,病例组的IL-6蛋白表达明显低于对照组;在蜕膜组织中,IL-6定位于蜕膜细胞和腺上皮细胞的细胞质内,且病例组蜕膜细胞的表达明显高于对照组,而2组在腺上皮细胞的表达无差异;H-E染色结果显示,病例组绒毛组织的滋养层变薄,细胞变性、坏死,细胞嗜酸性增强,绒毛中轴纤维化程度增强,蜕膜细胞失去细胞间连接,部分蜕膜细胞解体、核消失,细胞嗜酸性增强.结论:绒毛和蜕膜组织中IL-6蛋白表达的变化与反复自然流产有关,IL-6可能参与反复自然流产的病理过程.     

Bcl—2蛋白在正常、妊高征及胎儿宫内发育迟缓胎盘中的表达

目的 通过检测Bcl-2蛋白在正常各期胎盘、妊高征胎盘和胎儿宫内发育迟缓胎盘中的表达，探讨Bcl-2蛋白在胎盘的发育过程中的功能及妊高征和胎儿宫内发育迟缓的病理机制。方法 取正常的8－10周、20－22周、足月胎盘以及妊高征和胎儿宫内发育迟结的胎盘组织固定，石蜡包坦，用ABC法抗Bcl-2免疫组织化学染色，光镜观察。结果 正常早孕绒毛合体细胞滋养怪、细胞滋养层细胞核、绒毛吵轴细胞阳性，细胞滋养层的胞质、绒毛中轴基质阴性。正常中期和足月胎盘滋养层细胞、血管内皮细胞阳性。与正常足月胎盘相比，妊高征、台儿宫内发育迟缓胎盘几科不着色。结论 Bcl-2蛋白在正常各期胎盘中均有表达，而在妊高征和胎儿宫内发育迟缓胎盘几乎不表达，Bcl-2蛋白表达异常与妊高征与胎儿宫内发育迟缓的发病机制有关。     

自然流产模型小鼠蜕膜细胞凋亡及Bcl-2、Bax表达的研究

目的：通过比较正常妊娠模型小鼠及自然流产模型小鼠蜕膜细胞凋亡及Bcl-2、Bax蛋白的表达，从细胞及分子水平探讨自然流产的发病机制。方法：建立正常妊娠模型CBAXBALB／c和自然流产模型CBAXDBA／2。用免疫组化SABC法测定两组模型孕13天蜕膜细胞Bcl-2和Bax蛋白的表达，并通过MIAS-2000医用彩色病理图像免疫组化测量系统对其表达进行半定量分析，其结果用平均灰度值表示；同时应用DNA缺口原位末端标记技术（TUNEL）测定两组模型孕13天蜕膜细胞凋亡情况。结果：与正常妊娠模型相比，自然流产模型蜕膜细胞Bcl-2蛋白的表达降低（P〈0．01）；Bax蛋白的表达明显升高（P〈0．01）。蜕膜细胞凋亡指数，自然流产模型明显高于正常妊娠模型（P〈0．01）。结论：早孕期蜕膜组织细胞凋亡异常是自然流产的机制之一，Bcl-2／Bax途径可能是诱导早孕期蜕膜细胞凋亡的重要因素。     

CXCR2在不明原因自然流产患者绒毛和蜕膜组织中的表达

目的研究CXCR2在不明原因自然流产患者绒毛及蜕膜组织中的表达情况。方法应用免疫组织化学染色和图像分析技术,观察CXCR2在不明原因自然流产患者（病例组）和正常人工流产者（对照组）绒毛及蜕膜组织中的表达情况;HE染色后光镜下观察不明原因自然流产患者绒毛与蜕膜组织的形态学变化。结果免疫组化染色结果显示：两组CXCR2蛋白主要表达于绒毛的滋养层、蜕膜组织中的腺体,病例组绒毛滋养层和腺体CXCR2蛋白表达均强于对照组,两组比较,有显著性差异（P〈0.01）。病例组蜕膜细胞CXCR2蛋白呈阳性表达,对照组呈阴性;HE染色可见不明原因自然流产患者绒毛组织的滋养层变薄、滋养层细胞变性甚至坏死、滋养层嗜酸性增强、绒毛中轴纤维化程度增强;蜕膜组织中蜕膜细胞连接松散、蜕膜细胞空泡化、蜕膜组织中有大量炎细胞浸润。结论 CXCR2可能通过与其配体白细胞介素-8结合而参与不明原因自然流产的病理过程。     

白细胞介素-2在反复自然流产患者绒毛及蜕膜组织中的表达

目的:研究白细胞介素-2(IL-2)在反复自然流产患者绒毛及蜕膜组织中的表达水平及意义.方法:应用免疫组织化学和图像分析,分别对30例反复自然流产患者(病例组)和30例正常妊娠者(对照组)绒毛和蜕膜组织中IL-2的表达进行定位和半定量分析;应用显微镜观察反复自然流产患者绒毛和蜕膜组织的形态学变化.结果:免疫组织化学显色显示,IL-2蛋白定位于绒毛组织的滋养层细胞和蜕膜组织的蜕膜细胞细胞质内,且病例组的IL-2蛋白表达高于对照组.H-E染色结果显示,病例组的蜕膜组织结构模糊、细胞变性坏死、部分细胞连接消失,血管内皮有缺损;绒毛组织的滋养层变薄,细胞变性坏死,细胞嗜酸性增强,绒毛中轴纤维化程度增强.结论:绒毛和蜕膜组织中IL-2蛋白的高表达与反复自然流产有关,可能参与反复自然流产的病理过程.     

自然流产模型小鼠蜕膜细胞凋亡及相关基因的表达

目的 通过比较正常妊娠模型小鼠及自然流产模型小鼠蜕膜细胞凋亡及Bcl-2、Bax、Fas、FasL蛋白的表达,从细胞及分子水平探讨自然流产的发病机制.方法建立正常妊娠模型CBAXBALB/c和自然流产模型CBAXDBA/2.用免疫组化SABC法测定两组模型孕13 d蜕膜细胞Bcl-2、Bax、Fas、FasL蛋白的表达,并通过MIAS-2000医用彩色病理图像免疫组化测量系统对其表达进行半定量分析,其结果用平均灰度值表示;同时应用DNA缺口原位末端标记技术(TUNEL)测定两组模型孕13天蜕膜细胞凋亡情况.结果与正常妊娠模型相比,自然流产模型蜕膜细胞Bcl-2蛋白的表达降低(P＜0.01);Bax蛋白的表达明显升高(P＜0.01);FasL的表达明显升高(P＜0.01);Fas的表达两组比较无明显差异(P＞0.05).蜕膜细胞凋亡指数(AI),自然流产模型明显高于正常妊娠模型(P＜0.01).结论 早孕期蜕膜组织细胞凋亡异常是自然流产的机制之一,Bcl-2/Bax,Fas/FasL途径可能是诱导早孕期蜕膜细胞凋亡的重要因素.     

人胚胎生长发育过程中Bcl-2和Bax蛋白在食管组织中的表达

目的探讨人胚胎食管发育过程中食管壁细胞增殖与凋亡的变化规律,以及B细胞淋巴瘤/白血病-2(Bcl-2)和Bcl相关X蛋白(Bax)在食管生长发育中的表达意义。方法应用免疫组织化学ABC法检测第2、3、4月龄段人胚胎(共14例)食管壁组织内Bcl-2和Bax蛋白的表达。结果第2、3、4月龄段,人胚胎食管壁肌间神经丛、黏膜下神经丛和黏膜层内均有Bcl-2蛋白阳性表达,在食管上皮Bax蛋白呈弱阳性表达。结论Bcl-2和Bax细胞周期调控蛋白与人胚胎食管的生长发育关系密切。     

人胎发育过程中Bcl-2和Bax蛋白在小肠中的表达

目的 探讨人胎小肠发育过程中肠壁细胞增殖与凋亡的变化规律,及相关蛋白B细胞淋巴瘤/白血病-2(Bcl-2)和Bcl相关蛋白(Bax)在小肠生长发育中的表达意义.方法 应用免疫组织化学ABC法检测第2、3、4三个月胎龄段,人胎小肠肠壁组织内Bcl-2和Bax蛋白的表达.结果 第2、3、4三个月胎龄段,小肠肠壁肌间神经丛和黏膜下神经丛内均有Bcl-2阳性细胞分布,Bax在小肠黏膜层上皮细胞的细胞质内广泛表达.结论 Bcl-2/Bax蛋白在人胎小肠的生长发育过程中起重要的调节作用.     

Bcl-2和Bax蛋白在人胚胎早期脊髓发育中的表达及意义

为探讨人胚胎发育早期脊髓内神经细胞增殖与凋亡的变化规律及其相关蛋白Bcl-2和Bax在脊髓中的分布规律及其表达意义,本研究应用免疫组织化学ABC法检测了第2、3、4月胎龄段的人胚胎脊髓组织中央管、前角和后角处Bcl-2和Bax蛋白的表达状况。结果显示:在第2、3、4月胎龄段,脊髓中央管、前角和后角处均有Bcl-2和Bax蛋白阳性表达。本文结果提示,Bcl-2和Bax蛋白在人胚胎脊髓的生长发育过程中起重要的调节作用。     

Apoptosis in human chorionic villi and decidua in normal and ectopic pregnancy

To investigate possible effects of implantation on apoptosis, we examined the cleavage of DNA in human chorionic villi and decidua in intrauterine and ectopic pregnancy. Very limited but detectable cleavage of DNA was recognized in the chorionic villi and decidua in normal pregnancy. A ladder pattern, characteristic of the apoptotic breakdown of DNA, was present in the villi in tubal pregnancy. High molecular weight DNA was predominant in the decidua in tubal pregnancy. Quantitative analysis of low molecular weight fragments of DNA revealed a significant increase in the villous tissue, together with a significant decrease in the decidual tissue, in tubal pregnancy as compared to those in normal pregnancy. An analysis in situ revealed that apoptotic cells were predominant in the syncytiotrophoblast in tubal pregnancy. In decidual tissue, labelled cells were occasionally seen in normal pregnancy, and their numbers decreased in tubal pregnancy. The present study demonstrates that apoptosis occurs in the villi, but not in the decidua in tubal pregnancy, unlike the situation in normal pregnancy. Our results suggest that the implantation site might affect the occurrence of apoptotic changes in early pregnancy of humans.      

Localization of insulin-like growth factor-binding protein and endometrial beta-lactoglobulin in cultured decidual and chorionic villus cells

Immunofluorescence staining was used to localize two 'endometrial' proteins, insulin-like growth factor-binding protein and human beta-lactoglobulin homologue, in cultured cells of chorionic villi and decidua. Both cultured cell types were positive for each protein. These results indicate that immunohistochemical detection of these two proteins cannot be used to distinguish between cultured chorionic villus and decidual cells.     

Immunohistochemical localization of interferons in human placental tissues in normal, ectopic, and molar pregnancy

Interferon (IFN)alpha, beta, and gamma have been localized in normal and pathological human pregnancy using both polyclonal and monoclonal antibodies in immunohistochemical techniques. IFN alpha was localized to fetal chorionic villous syncytiotrophoblast throughout normal pregnancy, as well as to extravillous trophoblast in the placental bed and chorion lave. Maternal decidual leukocytes, as well as fetal Hofbauer cells in the villous mesenchyme, also contained IFN alpha, IFN gamma was detected in villous syncytiotrophoblast, while anti-IFN beta showed only patchy weak reactivity with syncytiotrophoblast. Reaction patterns on ectopic pregnancy tissues were similar to those in early intrauterine pregnancy. In molar pregnancy, reactivity for IFN alpha, beta, and gamma was observed in syncytiotrophoblast. Along with their potential anti-viral effects, placental interferons could play a role in local immunomodulation or in regulation of embryonic cell proliferation and differentiation.     

The effects of mifepristone (RU486) on prostaglandin dehydrogenase in decidual and chorionic tissue in early pregnancy

Prostaglandin dehydrogenase is the main inactivating enzymefor prostaglandins and therefore controls local levels of prostaglandins.Since there is some evidence that the expression of this enzymeis under progesterone control it is reasonable that one of theeffects of antiprogestin is to reduce the concentration of thisenzyme and thus increase the effective concentration of prostaglandinwithin tissue. We have investigated the amount of enzyme activitywithin decidua and chorionic villi from women receiving theantigestagen mifepristone (RU486) 12, 24 and 36 h prior to surgicalabortion, and examined the effect on tissue concentrations ofprostaglandin dehydrogenase. Women receiving mifepristone inall groups had a significant reduction in concentration of prostaglandindehydrogenase enzyme in decidual tissue. There was also a markedreduction in prostaglandin dehydrogenase in decidual cells followingRU486, as demonstrated by immunochemical methods. At this stageof pregnancy, prostaglandin dehydrogenase was present in abundancein cytotrophoblast cells of chorionic villi but virtually absentfrom syncytiotrophoblast. In chorionic villi after RU486 administrationin vivo, there were no obvious differences in prostaglandindehydrogenase distribution or reactivity in the majority ofcases.     

DEVELOPMENTAL REGULATION OF TISSUE TRANSGLUTAMINASE DURING HUMAN PLACENTATION AND EXPRESSION IN NEOPLASTIC TROPHOBLAST

The expression of tissue transglutaminase (tTG) was studied during the formation of the normal human placenta and in molar pregnancies and choriocarcinoma, in order to correlate its expression with the functional characteristics of the recognized trophoblast cell types. tTG expression was found to be developmentally regulated. Before 6–7 weeks' gestation, only the chorionic villous cytotrophoblast expresses tTG. Thereafter the overlying syncytiotrophoblast becomes positive. tTG expression is gradually downregulated in the intermediate trophoblast cells emerging from the tips of the chorionic villi invading the uterine tissue. In the decidual wall, the intermediate trophoblast does not express tTG, whereas scattered syncytial cells, the placental bed giant cells, express tTG. Villi from complete hydatidiform mole (CHM) show tTG expression in both the cyto- and the syncytiotrophoblast. The intermediate trophoblast cells from CHM show heterogeneous tTG expression, with a majority of negative cells, whereas extravillous syncytia always express tTG. In choriocarcinoma, the tumour cells show heterogeneous tTG expression, with a majority of positive cells. Analysis of tTG protein and mRNA in placental extracts by Western and Northern blotting did not provide evidence for expression of the truncated form of tTG found in some cell types. The regulated expression of tTG in the normal placenta suggests that the enzyme is involved in important trophoblastic functions and may participate in the control of invasion. © 1997 by John Wiley & Sons, Ltd.