白细胞介素-18在大鼠胚泡植入前后子宫的分布及表达

目的 系统研究了白细胞介素-18(IL-18)在大鼠胚泡植入前后子宫中的分布与表达,以探讨IL-18在胚泡植入过程中可能的生理作用.方法 免疫组织化学SP法,图像分析法.结果 IL-18在胚泡植入前后子宫均有表达,其主要分布于子宫内膜或蜕膜的腔上皮细胞、腺上皮细胞及子宫基质细胞或蜕膜细胞,子宫肌层及部分血管内皮细胞也有微弱表达.与非妊娠大鼠相比,妊娠大鼠子宫IL-18的表达明显增加(P<0.05).随着妊娠的进行,子宫IL-18的表达逐渐递增,并且IL-18在胚泡植人后期的表达显著高于植入前期(P<0.05).结论 IL-18可能参与胚泡植入,与妊娠的建立和维持有关,在妊娠中起重要作用.     

白细胞介素8在着床前小鼠子宫及胚卵的表达

目的:观察着床前小鼠子宫及胚卵白细胞介素8(IL-8)的表达.方法:免疫组织化学显色及图像分析技术,对IL-8蛋白在妊娠1～4 d小鼠子宫及胚卵的表达进行定位及半定量分析;用RT-PCR技术检测妊娠1～4 d小鼠子宫及胚卵IL-8mRNA的表达情况.结果:与未孕小鼠相比,妊娠各天小鼠子宫中IL-8蛋白和mRNA表达明显升高,于妊娠4d表达最强.IL-8蛋白和mRNA均在妊娠2 d(Ⅱ细胞期)的胚卵表达最弱,妊娠4d(胚泡期)的胚卵表达最强.结论:妊娠1～4 d小鼠子宫及胚卵持续表达IL-8蛋白和mRNA,尤其是在着床前表达量升高,提示它们可能参与小鼠胚泡的着床过程.     

CXCR2在妊娠早期小鼠子宫内膜的表达

目的研究CXCR2在妊娠早期小鼠子宫内膜的表达情况.方法应用免疫组织化学和图像分析技术,观察CXCR2在妊娠1d、4d、5d、6d小鼠子宫内膜的表达部位及蛋白水平的变化规律;HE染色后光镜下观察子宫内膜的形态学变化.结果免疫组化染色结果显示:在子宫内膜腔上皮,CXCR2于妊娠4d表达最强,妊娠5d开始逐渐下降,妊娠6d降至妊娠1d水平;在子宫内膜腺上皮和基质,CXCR2的表达水平随妊娠天数的增加而逐渐升高.HE染色可见妊娠1d子宫内膜腔上皮和基质中有大量中性粒细胞浸润;妊娠4d腔上皮中未见中性粒细胞,基质中则有大量中性粒细胞浸润;妊娠5d和妊娠6d腔上皮中未见中性粒细胞,基质中中性粒细胞数量甚少.结论CXCR2可能通过与其配体白细胞介素-8结合而参与小鼠胚泡着床过程.     

CRH样肽对大鼠胚泡着床的影响

本文对ＳＤ大鼠子宫内膜提取物作ＣＲＨ放射免疫分析，用免疫组织化学原位定位技术研究ＣＲＨ在大鼠子宫组织的分布，并采用自体对照的大鼠子宫角注射技术观察了ＣＲＨ抗血清对胚泡着床的影响，结果证实：（１）妊娠和非妊娠大鼠子宫内膜都含有ＣＲＨ样物质，其含量分别为：非妊娠大鼠３．６３±０．３２ｐｇ／ｍｇ湿组织（ｎ＝７）。妊娠大鼠与非妊娠大鼠间无显著差异。（２）ＣＲＨ样免疫活性物质存在子宫内膜固有层结缔组织的若干     

白细胞介素-8在培养大鼠肝癌细胞上的表达

白细胞介素－８（ＩＬ－８）是一种分布广泛的细胞因子，但其在肝癌细胞的定位尚未见报道。本研究采用免疫组织化学ＡＢＣ－ＧＤＮ法，对ＩＬ－８在培养的大鼠肝癌ＦＳＫ７９０２细胞系的分布进行了定位。结果显示，ＩＬ－８样免疫反应阳性物质分布于所有肝癌细胞，阳性物质为棕黑色颗粒状，颗粒均匀一致，主要集中分布于胞核周围的胞质中。ＩＬ－８样免疫反应阳性物质在肝癌细胞的分布并不完全一致，在未形成细胞单层的有突起的癌细胞中，ＩＬ－８样免疫反应阳性物质主要存在于胞核周围的胞质中，细胞突起的胞质中无ＩＬ－８样免疫反应阳性物质。本实验证明，肝癌细胞在不需要细胞因子刺激的情况下，可自发地表达ＩＬ－８（组成性表达）。     

白细胞介素—8在培养大鼠肝癌细胞上的表达

白细胞介素－８（ＩＬ－８）是一种分布广泛的细胞因子，但其在肝癌细胞的定位尚未见报道。本研究采用免疫组织化学ＡＢＣ－ＧＤＮ法，对ＩＬ－８在培养的大鼠肝癌ＦＳＫ７９０２细胞系的分布进行了定位。结果显示，ＩＬ－８样免疫反应阳性物质分布于所有肝癌细胞，阳性物质为棕黑色颗粒状，颗粒均匀一致，主要集中分布于胞核周围的胞质中。ＩＬ－８样免疫反应阳性物质在肝癌细胞的分布并不完全一致，在未形成细胞单层的有突起的癌细     

兔胚泡源性植入信号的在体研究模型及其初步研究

根据兔眼前房自体移植子宫内膜技术，建立了兔眼前房自体子宫内膜和异体３ｄ兔胚泡的双移植动物模型，并用该模型进行胚泡源植入信号的研究。结果表明，兔眼前房的子宫内膜受卵巢激素的调节。妊娠兔眼前房的子宫内膜受孕酮调节，呈分泌期子宫内膜。而非妊娠状态下，眼前房的子宫内膜受雌激素调节，呈增殖期状态。３ｄ胚泡易植入妊娠兔眼前房的子宫内膜，而难植入非妊娠兔眼前房的子宫内膜。胚泡植入到子宫内膜分泌细胞位置上，而不植     

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。     

白细胞介素-6 mRNA在内异症子宫内膜的表达变化

目的：探讨IL-6 mRNA与子宫内膜异位症发病的关系。方法：利用大鼠子宫内膜异位症（内异症）动物模型，采用逆转录聚合酶链反应（RT-PCR）技术，检测子宫内膜白细胞介素-6（IL-6）mRNA表达情况。结果：正常对照组，各时点间（2、4、6和8周）IL-6 mRNA表达无显著差异（P>0.05）。内异症模型组,在成模后（2、4、6和8周）其在位、异位子宫内膜IL-6 mRNA的表达呈增高趋势（P<0.01）。在各时点，内异症异位子宫内膜>内异症在位子宫内膜>正常子宫内膜IL-6 mRNA的表达（P<0.01）。结论：IL-6 mRNA可能与内异症的进展有关。IL-6 mRNA可能促进异位内膜的种植和生长。     

Expression of interleukin-8 receptors in endometriosis

BACKGROUND: Although the etiology of endometriosis is not well understood, chemokines and their receptors are believed to play a role in its pathogenesis. Therefore, we aimed to investigate the expression and localization of interleukin-8 (IL-8) receptors CXCR1 and CXCR2 in eutopic and ectopic endometrial tissues of women with endometriosis, and in endometrium of women without endometriosis. METHODS: Ectopic (n = 27) and homologous eutopic endometrium (n = 25) from women with endometriosis and endometrium from women without endometriosis (n = 27) were used for immunohistochemical analysis of CXCR1 and CXCR2. RESULTS: In normal endometrium, epithelial CXCR1 and CXCR2 immunostaining intensities were similar in the proliferative and secretory phase. Stromal CXCR1 expression was less then epithelial expression and did not show cyclical difference. No stromal CXCR2 expression was observed. In eutopic endometrium of women with endometriosis compared to endometrium of women without endometriosis, there was a significant increase in both proliferative and secretory phases for epithelial CXCR2 expression, and in proliferative phase for CXCR1 expression (P < 0.05). Both receptor immunoreactivities were significantly increased in the epithelial cells of ectopic endometrial tissues compared to that of normal endometrium (P < 0.05). CONCLUSIONS: These findings suggest that IL-8 and its receptors may be involved in the pathogenesis of endometriosis.     

Progressive rise in the expression of interleukin-6 in human endometrium during menstrual cycle is initiated during the implantation window

In order to be prepared for implantation, human endometriumundergoes a predictable series of proliferative and secretorychanges. Cytokines play an important role in regulation of thesechanges. Therefore, in this study, we immunolocalized the cytokine,interleukin-6 (IL-6), its receptor and the signal transducergp130 in human endometrium throughout the menstrual cycle. Duringthe entire menstrual cycle, the IL-6 receptor and gp130 werefound primarily in the endometrial glands and to a lesser extentin the stroma. The immunoreactivity of these proteins did notchange in endometrial cells during the entire menstrual cyclewith an exception of reduced immunoreactivity of gp130 in endometrialglands during menstrual phase. Immunostaining showed that immunoreactiveIL-6 was weakly expressed in human endometrium during the proliferativephase. Strong immunoreactivity for IL-6 appeared in endometriumduring the putative 'implantation window'. Expression was byfar most pronounced both in the glandular and surface epithelialcells. The amount of immunoreactive IL-6 in the epithelium progressivelyincreased during the secretory/menstrual phases. During thelate secretory phase, only stromal cells in the upper functionalisexhibited immunoreactivity for IL-6. Western blot analysis corroboratedthe immunohistochemical data. Human endometrial IL-6 consistedof a protein with an apparent mobility of 26 kDa. The immunoreactiveband of IL-6 was weak in the proliferative phase. The expressionof this protein increased progressively during the secretory/menstrualphases. The findings show a cell-specific pattern of distributionfor immunoreactive IL-6 in human endometrium. The menstrualcycle-dependent expression of IL-6 suggests that this cytokinemay play a role in changes in endometrium that prepare thistissue for implantation and menstrual shedding.     

ATF4基因在小鼠胚胎围着床期子宫内膜的表达

目的 探讨转录激活因子4(ATF4)在小鼠胚胎围着床期子宫内膜中的表达规律及在胚胎植入过程中的作用.方法 妊娠0～7d小鼠子宫内膜,用RT-PCR、免疫组织化学法和Western blot分别检测小鼠内膜ATF4 mRNA及蛋白的表达;用子宫角内注射ATF4抗体进行干预.结果 1)妊娠各组小鼠子宫内膜ATF4 mRNA的表达量均显著高于未孕组d0(P ＜0.0S),且随妊娠天数的增加其表达量逐渐增高,到d4达到峰值,之后逐渐下降;2)ATF4蛋白主要在基质细胞胞质表达,其表达规律与mRNA表达规律基本一致.3)子宫角ATF4抗体注射后与对照组相比着床数明显减少.结论 ATF4在小鼠妊娠早期可能参与子宫内膜蜕膜化过程,有利于着床.     

E-选择素在胚胎着床过程中小鼠子宫内膜的表达规律

目的：研究E-selectin在胚胎着床及早期分化中的作用。方法：用RT—PCR、Immunoblot、Western blot法检测小鼠胚胎着床前后蜕膜组织E-selectin的表达。结果：E-选择素在孕4天的小鼠子宫内膜中的表达出现升高，第5天达到峰值，随后妊娠第6、第7天E-selectin蛋白的表达则逐渐下降至正常水平。结论：提示E-选择素可能参与了胚胎着床过程。     

Symposium: reproduction in baboons. From blastocyst to placenta: the morphology of implantation in the baboon

Implantation and placentation in the baboons share many morphologicalfeatures with other primates, as well as having some specificdistinctions. The ability to use deturgescence of the sex skinas a method of timing ovulation and the ease with which theuterine lumen can be flushed have been used to examine morphologicalaspects of blastocyst differentiation and implantation in thisspecies. Preimplantation blastocysts were obtained by non-surgicalflushing of the uterus 68 days after ovulation, and implantationsites were excised from uteri removed on days 10-16 of gestation.All tissues were prepared for electron microscopy by aldehydefixation and plastic embedding. Maturation of trophoblast fromthe compacted morula stage to the expanded blastocyst stageincludes increase in numbers of polyribosomes, changes in conformationof mitochondria, and development of an effective endocytic apparatus.An endodermal layer forms beneath the inner cell mass priorto loss of the zona pellucida, and parietal endodermal cellsextend beyond the inner cell mass. Azonal blastocysts have regionsof syncytial trophoblast adjacent to the inner cell mass, andthey may represent adhesion stages of early implantation. Inearly postimplantation stages, trophoblast replaces the uterineepithelium and processes of syncytial trophoblast invade dilatedsuperficial maternal vessels. In subsequent lacunar stages thereis rapid elevation of the developing conceptus above the uterinesurface as the lacunae enlarge. Cytotrophoblast rapidly entersmaternal vessels, and arterioles are partially or completelyoccluded by migrating cytotrophoblast. The early access to controlledmaternal blood flow apparently allows trophoblastic lacunaeto expand superficially as opposed to more extensive endometrialinvasion.     

Characterization of IK cytokine expression in mouse endometrium during early pregnancy and its significance on implantation

The expression of IK cytokine was investigated in the mouse endometrium during early pregnancy (D1-D7 of pregnancy) and pseudopregnancy using real-time PCR, western blotting and immunohistochemical analysis, and the effects of IK cytokine on embryo implantation were observed by injection with antisense IK cytokine oligodeoxynucleotides in the uterine horn. Our data showed that the expression of IK cytokine mRNA increased gradually from D1 to D4 of pregnancy and reached a peak level at D4 of pregnancy (P<0.05). Western blotting and immunohistochemical analysis revealed that the expression of IK cytokine protein increased gradually from D1 to D5 of pregnancy and reached a peak level at D5 of pregnancy (P<0.05). The expression of IK cytokine in the pseudopregnant uterus was significantly lower compared to that in the normal pregnant uterus and the level of the protein never showed a high peak during the whole pseudopregnancy. The expression of IK cytokine at the implantation site was much stronger than that in the peri-implantation site on Day 5 of pregnancy. After 24 and 48 h of injection with antisense IK cytokine oligodexynucleotides in the uterine horn on D3 of pregnancy (i.e. implantation window), the expression of IK cytokine in the uterus was remarkably inhibited, while the expression of major histocompatibility complex II (MHC II) increased and the number of implanted embryos significantly decreased in the site of uterine horns receiving antisense IK cytokine (P<0.05). These results suggested that IK cytokine may play a crucial role in implantation.     

Hormonal and embryonic regulation of chemokine receptors CXCR1, CXCR4, CCR5 and CCR2B in the human endometrium and the human blastocyst

Chemokines are implicated in the implantation process. The aim of this study was to investigate mRNA expression and protein levels of chemokine receptors CXCR1, CXCR4, CCR5 and CCR2B in human endometrium throughout the menstrual cycle, during HRT and in the human blastocyst. The regulation of chemokine receptors in the endometrial epithelium was also studied using an in-vitro model for the apposition phase of human implantation. We found up-regulation of endometrial CXCR1 mRNA (419-fold increase), CCR5 mRNA (612-fold increase) and CCR2B mRNA (657 fold-increase) during the luteal phase peaking in the pre-menstrual endometrium. CXCR4 mRNA levels presented a specific although modest (18-fold increase) up-regulation during the implantation window. These findings were corroborated at the protein level in natural and HRT cycles. Immunoreactive CCR5 and CCR2B receptors were detected in human blastocysts whereas CXCR4 and CXCR1 were not present. Chemokine receptors in cultured endometrial epithelial cells showed an up-regulation and polarization of CXCR1, CXCR4 and CCR5 receptors when a human blastocyst was present. The specific distribution and regulation of chemokine receptors in the endometrial epithelium and the human blastocyst suggest a possible implication of these receptors in the apposition and adhesion phases of human implantation.     

降钙素对体外培养的小鼠胚泡发育、分化的影响

目的：探讨降钙素(CT)与小鼠胚泡发育及植入的关系。方法：将孕D4小鼠胚泡随机分组并接种到添加不同成分的培养液内进行体外培养。于培养24、48h后在倒置显微镜下观察、记录胚泡的孵化、粘附及扩展生长情况。结果：培养24h后,添加10nmol/L CT组与联合培养组胚泡的粘附率分别为31．71％、32．35％,和对照组(16．30％)相比,均有明显增加。培养48h后,添加10nmol/L CT组与联合培养组胚泡的孵化率、粘附率、扩展率均比对照组有非常显著增高。结论：CT可明显地促进体外培养的小鼠胚泡孵化、粘附及扩展生长,与植入过程密切相关。     

An in-vitro model for stromal invasion during implantation of the human blastocyst

BACKGROUND: Implantation failure is likely to be a major cause of infertility. Studies in mice have identified a number of molecules that are involved in implantation, but the mechanisms of implantation in the human remain unclear, largely due to the lack of models for implantation in the human that provide functional information. METHODS: Human hatched blastocysts were co-cultured with human endometrial stromal cell monolayers. Time-lapse photography of implanting blastocysts, immunostaining for cytokeratin and actin, and measurement of hCG secreted into the culture supernatants were performed. RESULTS: Blastocysts attached to and implanted into the stromal cell layer. Trophoblast outgrowth onto, and invasion into, the stromal cell layer occurred largely at two opposite poles, the orientation of which was aligned to that of the stromal cell fibroblasts. High-resolution image analysis demonstrated that the trophoblast completely penetrated the stromal cell layer. Immunostaining of whole-mounts of implantation sites revealed distinctive actin and cytokeratin-positive anchoring structures adjacent to the basal surface of the trophoblast. Blastocysts implanting into stromal cells secreted higher levels of hCG compared with those cultured on plastic. CONCLUSIONS: A robust model for the study of mechanisms of implantation of the human embryo into the endometrial stroma has been established.     

Molecular aspects of implantation: The signals and molecular pathways involved in implantation, a symbiotic interaction between blastocyst and endometrium involving adhesion and tissue invasion

Implantation is a complex process requiring the interactionof the blastocyst, and subsequently the developing embryo withthe endometrium. Initially, the detailed cellular interactionsimplicated in this process were defined. More recently, manysignals and molecular pathways are recognized that induce, orregulate the complex series of interactions required for implantation.In this review, the cellular and molecular interactions thattake place during implantation are discussed.