白细胞介素-8在妊娠早期小鼠子宫内膜的表达及对胚泡着床的影响

目的：研究白细胞介素-8（IL-8）在妊娠早期小鼠子宫内膜的表达及其对小鼠胚泡着床的影响。方法：用免疫组化法及图像分析技术对IL-8在妊娠1～6d小鼠子宫内膜的表达进行定位和测定;子宫角注入IL-8抗体,探讨IL-8对胚泡着床的影响。结果：在子宫内膜腔上皮,IL-8主要定位于细胞的游离面,妊娠1d阳性反应最弱,4d表达最强,5d有所下降,6d又开始增强;在子宫内膜腺上皮,妊娠4d表达最弱,5d和6d最强;在子宫内膜的基质细胞,随妊娠天数增加,IL-8的表达逐渐增强。IL-8Ab明显抑制小鼠胚泡着床。结论：IL-8在妊娠早期小鼠子宫内膜持续表达,并参与胚泡的着床调控过程。     

层粘连蛋白在早孕小鼠子宫内膜中的表达

层粘连蛋白 (ＬａｍｉｎｉｎＬＮ)是细胞外基质中重要的大分子非胶原糖蛋白 ,由座落于基膜上的上皮细胞、内皮细胞及被基膜包绕的肌细胞、垂体细胞合成并分泌贮存于基质中。研究表明 ,ＬＮ与细胞膜表面受体结合后可产生多种生物学作用 ,如介导上皮细胞生长、增殖、分化 ,促进细胞粘附、辅展、移动 ,并在肿瘤细胞转移过程中起重要作用。有人曾用间接免疫荧光方法对小鼠早期胚胎进行研究 ,发现ＬＮ在桑椹胚和内细胞团及其滋养层均有较强的阳性反应 ,在外胚层与内胚层之间的细胞外基质中可见ＬＮ呈条带状荧光。但有关胚泡植入前后子宫内膜中Ｌ…     

层粘连蛋白和纤维粘连蛋白在着床期小鼠子宫内膜中的表达

目的：为探讨层粘连蛋白和纤维粘连蛋白在胚泡着床过程中的生物学作用。方法：本实验使用昆明雌性小鼠30只,分为未孕和早期妊娠共6组。通过免疫组织化学方法,检测了层粘连蛋白和纤维粘连蛋白在着床期小鼠子宫内膜中的分布状况。结果：胚泡着床开始时（即受精后4－5天）,层粘连蛋白和纤维粘连蛋白均出现一个表达高峰,尔后纤维粘连蛋白迅速减少。受精后6－8天,细胞外基质中的层粘连蛋白逐渐增加。结论：层粘连蛋白和纤维粘连蛋白在胚泡着床微环境和植入调节中发挥重要作用。     

小鼠子宫内膜中鸡冠珊瑚树凝集素和双花藕豆凝集素受体在胚泡植入过程中的表达

目的 探讨小鼠子宫内膜鸡冠珊瑚树凝集素(ECL)受体和双花藕豆凝集素(DBA)受体与胚泡植入的关系.方法 以生物素标记的ECL和DBA来检测怀孕侧和输卵管结扎未孕侧小鼠孕早期子宫内膜中两种凝集 素受体的分布状况和变化规律.结果怀孕侧,ECL主要表达于胚胎滋养层细胞及其基膜,蜕膜细胞及其周围的细胞外基质(ECM);未孕侧,主要表达于子宫内膜上皮和腺上皮.在怀孕侧,DBA受体主要表达于胚胎滋养层细胞和子宫内膜血管基膜;在未孕侧,主要表达于子宫内膜血管的基膜.结论 ECL和DBA受体与小鼠胚泡植入过程密切相关.     

小鼠胚泡植入早期FucT-Ⅶ和sLe(X)寡糖抗原表达

目的探讨妊娠第1～5天(d1～5)小鼠子宫内膜唾液酸化的路易斯寡糖[sLe(X)]合成关键酶α-1,3岩藻糖基转移酶基因(FucT-Ⅶ)的表达和其表面sLe(X)寡糖抗原表达对胚胎植入的影响。方法应用RT-PCR和免疫组化方法检测妊娠第1～5天小鼠子宫内膜FucT-Ⅶ及寡糖抗原的表达规律。用子宫角内注射sLe(X)单克隆抗体的方法检测小鼠胚胎着床数。结果妊娠第1～5天小鼠子宫组织均有FucT-Ⅶ的表达,且在妊娠d4表达更强(P<0.05)。妊娠第1～5天sLe(X)寡糖抗原表达在子宫内膜的腔上皮和腺上皮。单侧子宫角sLe(X)抗体注射后胚胎的着床数量明显较对侧(注射等体积生理盐水)减少。结论在小鼠胚胎着床过程中,FucT-Ⅶ及sLe(X)寡糖抗原可能参与了胚泡的定位、黏附及侵袭过程,在胚泡植入的早期发挥作用。     

小鼠胚胎与子宫内膜中整合素α_5的表达

目的:观察整合素α5在小鼠早期胚胎和子宫内膜中的表达分布状况,探讨整合素在胚胎着床中的可能作用。方法:用免疫组织化学ABC法染色,对发育不同时期的小鼠胚卵和妊娠2 d～6 d、8 d、13 d以及非妊娠期的子宫内膜中整合素α5的表达分布进行形态学观察与半定量分析。结果:整合素α5的阳性反应物质在小鼠早期胚卵胞浆内;非妊娠期子宫内膜和妊娠期的脱膜上皮与基质细胞内均有整合素α5表达,植入前表达相对较弱,植入后随妊娠的进展逐渐增强。结论:提示整合素α5在小鼠胚卵着床中起重要作用:推测它不参与胚卵对母体子宫内膜的识别和粘附,而是参与了粘附后的侵入及胚胎发育过程。     

T淋巴细胞侵袭转移诱导蛋白1在小鼠子宫内膜基质细胞-胚泡共培养体系中对胚泡黏附的影响

目的:检测不同浓度T淋巴细胞侵袭转移诱导蛋白1 (Tiaml)抗体下,小鼠着床窗子宫内膜基质细胞内的Tiaml蛋白表达及其对基质细胞-胚泡共培养体系中胚泡黏附的影响,探讨Tiaml对小鼠胚胎着床的影响.方法:提取着床窗小鼠子宫内膜基质细胞并构建基质细胞-胚泡共培养体系;分别用免疫细胞化学和免疫印迹法检测不同浓度抗Tiaml抗体时着床窗基质细胞Tiaml蛋白的表达及其对胚泡黏附的影响.结果:抗体干预后着床窗基质细胞内的Tiaml蛋白表达水平降低;在不同抗体浓度下胚泡黏附率有差异.结论:随Tiaml抗体浓度增加,着床窗小鼠子宫内膜基质细胞Tiaml蛋白表达量及胚泡黏附率降低.表明Tiarnl可能在胚泡植入过程起重要的作用.     

妊娠早期小鼠卵巢、输卵管及子宫内诱导型一氧化氮合酶的分布

目的　了解胚泡着床前后妊娠昆明系小鼠卵巢、输卵管及子宫内诱导型一氧化氮合酶 (inducible ni-tric oxide synthase,i NOS)的分布。　方法　免疫细胞化学 L SAB法。　结果　妊娠 2～ 5 d小鼠的卵巢内 ,黄体细胞上有 i NOS的阳性表达 ;输卵管粘膜上有 i NOS的分布 ,肌层则为阴性 ;妊娠 2、3d的小鼠子宫内 ,阳性标记主要出现在子宫内膜上皮以及子宫内膜中的子宫腺上皮 ,内膜基质细胞为阴性 ;妊娠 4d的子宫内 ,子宫腺及蜕膜部分均有 i-NOS的分布 ,妊娠 5 d时 ,在小鼠胚泡的表面也检测到了 i NOS的存在。　结论　小鼠胚泡着床前后 ,在其卵巢、输卵管及子宫内均有 i NOS的存在 ,提示 i NOS在小鼠胚胎早期发育及着床过程中起作用。     

小鼠胚胎与子宫内膜中整合素α5的表达

目的：观察整合素α5在小鼠早期胚胎和子宫内膜中的表达分布状况，探讨整合素在胚胎着床中的可能作用。方法：用免疫组织化学ABC法染色，对发育不同时期的小鼠胚卵和妊娠2d-6d、8d、13d以及非妊娠期的子宫内膜中整合素内的表达分布进行形态学观察与半定量分析。结果：整合素α5的阳性反应物质在小鼠早期胚卵胞浆内；非妊娠期子宫内膜和妊娠期的脱膜上皮与基质细胞内均有整合素α5表达，植入前表达相对较弱，植入后随妊娠的进展逐渐增强。结论：提示整合素α5在小鼠胚卵着床中起重要作用：推测它不参与胚卵对母体子宫内膜的识别和粘附，而是参与了粘附后的侵入及胚胎发育过程。     

体外研究整合素α5对小鼠胚泡着床的作用

目的　在人蜕膜细胞与小鼠胚泡共培养过程中 ,研究整合素α5(integrinα5)在小鼠胚泡着床中的作用 ,分析其作用机制。　方法　利用人蜕膜细胞与小鼠胚泡共培养模型 ,模拟胚泡在体内的着床过程 ,加入integrinα5抗体及其配体纤粘连蛋白 (FN)抗体 ,对胚泡的粘附、外延生长、着床率进行形态学观察和统计学分析。　结果　人子宫蜕膜经分离纯化后 ,检测活细胞比例 >92 % ,贴壁的蜕膜细胞比例 >95 % ,小鼠胚泡可在蜕膜上粘附、生长 ;integrinα5抗体及配体FN的抗体均能抑制小鼠胚泡的侵入、外延生长 ,但不抑制胚泡的孵出和粘附。　结论　建立了小鼠体外着床模型 :integrinα5在着床中促进小鼠胚泡的侵入与铺展 ,但对胚泡孵出、子宫内膜的识别与粘附不产生影响。     

宫腔注射纤连蛋白抗体和层粘蛋白抗体对小鼠胚泡着床的影响

目的：主要研究纤连蛋白抗体和层粘连蛋白抗体对胚泡着床的影响,方法：取孕４天小鼠,分别在两侧子宫角内注射纤连蛋白及层粘连蛋白抗血清,于孕８天检查胚胎床数,取子宫观察内膜的组织学及免疫组织化学变化。结果：纤维蛋白抗血清及层粘连蛋白抗血清均有显著抑制胚泡着床的效应,组织学及免疫组化结果表明,纤连蛋白抗血清及层粘连蛋白抗血清可引起蜕膜细胞退化解体,拮抗纤维蛋白及层粘连蛋白在脱膜组织中的表达,结论：纤连蛋白     

Penetration of the uterine epithelial basement membrane during blastocyst implantation in the mouse.

For many species, blastocyst implantation is associated with a reduction in the number of cellular and extracellular matrix layers which separate the trophoblast from maternal vasculature. Following loss of uterine epithelial cells along the distal mural trophoblast, the mouse blastocyst encounters the residual epithelial basement membrane. This sheet of extracellular matrix must be breached and later removed prior to trophoblast invasion of the uterine stroma and formation of the placenta. The interactions between the trophoblast, luminal epithelial basement membrane, and decidual cells during the time when embryonic and uterine stromal cells first achieve contact were examined in this study. Distal mural trophoblast of activated delay blastocysts was in contact with the residual luminal epithelial basement membrane 36 hr after estrogen administration. This portion of the basement membrane contained areas in which the usual linear appearance was changed to an irregular, tortuous profile. The lamina densa frequently appeared flocculent and diffuse. Cytoplasmic processes from trophoblast and decidual cells simultaneously perforated the basement membrane at multiple discrete loci. With further development the basement membrane was lost, leaving trophoblast and decidual cells in close contact over large areas. In normally implanting blastocysts a similar stage of embryonic development, as described above, was attained by 0400 hr on day 6 of pregnancy. Regions of convoluted epithelial basement membrane were also seen in these implantation sites. However, only decidual cell processes were seen penetrating the residual basement membrane. These processes extended to the fetal side of the basement membrane and separated that matrix from overlying trophoblast. They contained organelles and formed rudimentary intercellular junctions with the trophoblast.(ABSTRACT TRUNCATED AT 250 WORDS)     

Penetration of the uterine epithelial basement membrane during blastocyst implantation in the mouse

For many species, blastocyst implantation is associated with a reduction in the number of cellular and extracellular matrix layers which separate the trophoblast from maternal vasculature. Following loss of uterine epithelial cells along the distal mural trophoblast, the mouse blastocyst encounters the residual epithelial basement membrane. This sheet of extracellular matrix must be breached and later removed prior to trophoblast invasion of the uterine stroma and formation of the placenta. The interactions between the trophoblast, luminal epithelial basement membrane, and decidual cells during the time when embryonic and uterine stromal cells first achieve contact were examined in this study. Distal mural trophoblast of activated delay blastocysts was in contact with the residual luminal epithelial basement membrane 36 hr after estrogen administration. This portion of the basement membrane contained areas in which the usual linear appearance was changed to an irregular, tortuous profile. The lamina densa frequently appeared flocculent and diffuse. Cytoplasmic processes from trophoblast and decidual cells simultaneously perforated the basement membrane at multiple discrete loci. With further development the basement membrane was lost, leaving trophoblast and decidual cells in close contact over large areas. In normally implanting blastocysts a similar stage of embryonic development, as described above, was attained by 0400 hr on day 6 of pregnancy. Regions of convoluted epithelial basement membrane were also seen in these implantation sites. However, only decidual cell processes were seen penetrating the residual basement membrane. These processes extended to the fetal side of the basement membrane and separated that matrix from overlying trophoblast. They contained organelles and formed rudimentary intercellular junctions with the trophoblast. It is concluded that decidual cells play an active role in the penetration of the epithelial basement membrane and may aid in its disintegration. © 1992 Wiley-Liss, Inc.     

Experimental studies on the initial trophoblast endometrial interaction.

6.1. Attachment and penetration of the uterine epithelium in the human: The present studies describe to our knowledge the only studies of human in vitro attachment of blastocysts to uterine epithelial monolayers. From these studies, using other mammalian species as controls, it is concluded that initial attachment and penetration of the epithelium in the human might be of the intrusive type also seen in the Rhesus monkey (Lindenberg et al, 1986; 1989; Enders, 1972; 1976). This implicates an intensive and intimate cell-cell recognition and interaction already at the stage of epithelial attachment of the human embryo. 6.2. Oligosaccharide determinants in implantation: In the mouse we have identified an oligosaccharide Lacto-N-fuco-pentaose I, which inhibits blastocyst attachment to uterine epithelial cells in vitro. Monoclonal antibodies recognizing this epitope have identified LNF I-like determinants in the secretion and on the surface of the mouse endometrial epithelium during early pregnancy. LNF I-HSA/BSA-FITC conjugates have identified a receptor for LNF I on the surface of the blastocyst at the time of implantation. Furthermore, this study demonstrates a redistribution of these determinants prior to the day of implantation, supporting the hypothesis, that LNF I on the surface of the uterine lumen may contribute to the recognition and attachment even during the initiation of implantation. Component(s) carrying LNF-1 in the secretion might be regulatory molecules helping to secure the proper time for implantation in the mouse. 6.3: Future aspects. Whether the proposed mechanism of blastocyst-endometrium adhesion can be applied to species other than the mouse has still to be determined, but there are indications that this is worth testing.(ABSTRACT TRUNCATED AT 250 WORDS)     

Molecular aspects of implantation: Blastocyst--endometrial interaction: an appraisal of some old and new ideas

The nature of the early interaction between blastocyst and endometrialepithelium is known to be highly specific within individualspecies. Despite this unique initial interaction, the trophoblastpromptly establishes a common theme as invasive cells penetratethe endometrium and colonize its local blood vessels. In allanimals with this type of implantation the blastocyst playsa more active role than the endometrial epithelium. The aggressivebehaviour of the blastocyst may be induced by signals from theendometrium which has been primed with preimplantation ovariansteroids. Activation of the blastocyst may reflect triggeringof synchronized paracrine loops, activated during the implantationwindow. Endometrial cytokines and eicosanoids are probably theprimary signals that drive the interlinking paracrine loopsand are essential for decidualization, trophoblast growth andinvasion. Since the dominant feature of early implantation israpid trophoblast migration, particularly in primates, the degreeto which the blastocyst attaches to the apical surface of theluminal endometrial epithelium during implantation is uncertain.The thickness of the glycocalyx on the uterine luminal epitheliumduring the peri-implantation period varies considerably betweenspecies. Its role in blastocyst attachment, if any, and in trophoblastcell locomotion, requires further study. The molecular propertiesand functions of the uterine epithelial plasma membrane, andthose of the interacting trophoblast at the site of implantation,have been largely neglected and require further extensive study. blastocyst adhesion/endometrial receptivity/implantation/plasma membrane/trophoblast     

凝集素PHA-E、UEA受体在真孕小鼠和假孕模型小鼠孕早期子宫内膜中的变化

目的：探讨子宫内膜凝集素PHA-E和UEA的受体与胚泡植入的关系。方法：应用生物素标记的凝集素PHA-E和UEA，来检测真孕组、单纯假孕组和蜕膜化假孕组小鼠孕早期子宫内膜中两种凝集素受体的分布状况和变化规律。结果：PHA-E受体广泛分布于各组小鼠子宫内膜腔上皮和腺上皮、蜕膜细胞表面及其周围的ECM；其在真孕组的水平均显著高于假孕组，且在孕9d时，蜕膜化假孕组的水平又显著高于单纯假孕组。UEA受体则主要分布于子宫内膜腺上皮游离缘；其在真孕组的水平高于单纯假孕组，在孕6、9d时，真孕组的水平又明显低于蜕膜化假孕组。结论：凝集素PHA-E受体与小鼠胚泡植入过程密切相关，对UEA受体与胚泡植入的关系尚难做出满意的解释。     

凝集素DBA和WGA的受体在生殖周期小鼠子宫内膜中的亲合细胞化学

目的：探讨凝集素DBA和WGA的受体在胚泡着床过程中的生物学作用。方法：取生殖周期不同阶段的小鼠子宫内膜组织，采用亲合细胞化学和图像分析方法，检测WGA和DBA的受体在子宫内膜中的分布和变化情况。结果：以上2种凝集素受体均存在于生殖周期不同阶段的小鼠子宫内膜，但2者的存在部位和数量有差异。WGA受体广泛存在于子宫内膜腔上皮和腺上皮的游离缘，以及孕期胚胎组织和蜕膜细胞表面及ECM，DBA受体仅见于子宫内膜或胚胎组织血管的内皮及其基膜。并且2种受体在孕早期，尤其是围植入期的水平均显著高于间情期和哺乳期的水平。结论：凝集素WGA和DBA的受体与小鼠胚泡植入过程相关。     

Characteristics of the uterine luminal surface epithelium at preovulatory and preimplantation stages in the marmoset monkey

Light and electron microscopy was used to examine the apical luminal epithelial surface of the uterus at preovulatory and preimplantation stages in the marmoset monkey. Luminal surface charge, detected by cationic ferritin staining, progressively decreased from preovulation to day 11 of pregnancy. The smooth, regular apical plasma membrane at preovulatory stages was in contrast to the convoluted, irregular surface observed during early pregnancy, especially at 1 day before blastocyst implantation. Profiles of microvilli were also altered, becoming thicker and more irregular during early pregnancy. Within the epithelial cell body, cyclic morphologic changes were seen, largely in association with secretory organelles. Giant phagocytic bodies were prominent at all stages examined, although their composition and intensity of staining varied throughout the cycle. Weak to moderate estrogen alpha and progesterone receptor immunostaining of the luminal epithelium was found during preovulatory and early pregnancy stages. This study describes complex cyclic changes in the morphology and biochemical make-up of the uterine luminal epithelial surface in a New World monkey in preparation for blastocyst attachment.