细胞间粘附分子-1在小鼠早孕期子宫内膜的表达规律及调节的研究

为研究细胞间粘附分子 1 (ICAM 1 )在孕早期的作用 ,本文观察了孕早期小鼠子宫内膜 ICAM 1蛋白水平的表达规律及细胞因子 L IF、IL 1、GM CSF和 IL 6对孕 d 4子宫内膜 ICAM 1表达的调节。方法 :1 .收集孕 1～ 8d小鼠子宫内膜 ;2 .将孕鼠随机分为实验组和对照组 ,在孕 d2～ 4分别腹腔注射不同的细胞因子或 PBS,d4收集子宫内膜。采用 Western blot方法测定子宫内膜 ICAM 1的蛋白表达水平。结果显示 :ICAM1呈时序性表达 ,孕 d1表达量较低 ,d2开始升高 ,到 d4达高峰 ,此后缓慢下降。L IF、IL1 β、IL 6、GM CSF对子宫内膜 ICAM 1的表达有促进作用。结论 :ICAM 1在围着床期表达水平较高 ,可能参与了胚胎着床。细胞因子 L IF、IL 1、IL 6和 GM CSF对其表达有促进作用。     

控制性卵巢刺激中黄体期添加雌激素对小鼠子宫内膜组织胞饮突变化和白血病抑制因子表达的影响

目的 研究控制性卵巢刺激(COS)中黄体期添加雌激素对子宫内膜容受性的影响,探讨COS中不同黄体期支持方案对小鼠子宫内膜中胞饮突发育情况、白血病抑制因子(LIF)表达的影响.方法 将正常雌性昆明小鼠随机分为A组(单纯COS)、B组(COS+黄体酮)、C组(COS+黄体酮+雌激素)和D组(自然周期,未应用药物)共4组,扫描电镜下观察各组小鼠妊娠第3～5天子宫内膜胞饮突的发育情况;采用免疫组化方法 检测LIF蛋白在各组小鼠妊娠第3～5灭子宫内膜中的表达情况.结果 (1)B、C、D组小鼠妊娠第3天子宫内膜出现发育中的胞饮突,妊娠第4天出现完全发育的胞饮突,妊娠第5天出现退化的胞饮突;A组小鼠妊娠第3天可见少量完伞发育的胞饮突,妊娠第4天可见退化的胞饮突.(2)妊娠第3～5大小鼠子宫内膜LIF蛋白表达水平的平均值,C组(138.5±20.3)、D组(143.1±19.0)均明显高于A组(103.2±5.0),差异均有统计学意义(P<0.05),C、D组妊娠第4天LIF蛋白表达均最强,呈强阳性;B组表达水平(123.5±10.8)高于A组,但低于C、D组,差异也均有统计学意义(P<0.05),B组妊娠第4天LIF蛋白表达最强,呈阳性;A组小鼠妊娠第3天LIF蛋白表达最强,呈弱阳性.A组妊娠第3天小鼠与B组、C组和D组妊娠第4天小鼠子宫内膜中LIF蛋白表达水平之间两两比较,差异均有统计学意义(F=55.76,P<0.01).结论 COS中黄体期支持时添加雌激素能改善小鼠子宫内膜中胞饮突的发育及LIF蛋白的表达,从而增加子官内膜容受性.     

白血病抑制因子在子宫内膜异位症并发不孕患者黄体中期子宫内膜中的表达

目的:探讨白血病抑制因子(LIF)在子宫内膜异位症(EM)导致不孕机制中的作用。方法:采用原位杂交及免疫组织化学方法对15例异位症并发不孕患者黄体中期子宫内膜中的LIF mRNA及其蛋白的表达进行定位及半定量分析,并以15例有生育妇女作为对照组。结果:子宫内膜异位症组LIF mRNA平均光密度为3.12±0.32,LIF蛋白平均光密度为9.31± 2.10,显著低于对照组的6.25±2.14与15.70±1.52(P<0.05)。结论:子宫内膜异位症组黄体中期子宫内膜LIF表达下降可能是其不孕的原因之一。     

肿瘤转移抑制因子CD82/KAI1基因在小鼠子宫内膜的动态表达

目的：探讨肿瘤转移抑制因子CD82/KAI1在小鼠胚胎着床过程中的功能。方法：应用RT-PCR和免疫组织化学技术分别观察CD82/KAI1mRNA和蛋白在小鼠动情周期和早孕子宫中的表达。结果：在动情周期中,CD82/KAI1mRNA除在动情期较低外,其余三期的表达量基本相同,但蛋白质却是在动情期表达最丰富,子宫内膜上皮和基质均呈阳性。在早孕子宫中,CD82/ KAI1mRNA的表达逐渐增多,蛋白质表达的量和范围也逐渐增强。结论：CD82/KAI1在小鼠子宫中呈动态表达,提示它在胚胎精确侵袭子宫内膜的调节中发挥作用。     

白血病抑制因子在子宫内膜、早孕蜕膜及绒毛组织中的表达

白血病抑制因子 (ＬＩＦ)在哺乳动物子宫内膜中是胚泡成功植入的关键因素之一[1] 。本研究应用逆转录聚合酶链反应 (ＲＴ ＰＣＲ)技术 ,对ＬＩＦｍＲＮＡ在人子宫内膜、早孕蜕膜及绒毛组织中的表达进行定量分析 ,探讨其在受精卵着床及早期胚胎维持中的作用。一、资料与方法1 资料来源 :( 1)正常子宫内膜 :选取 1998年 7月至 1999年 9月在我中心因子宫肌瘤行子宫切除的病人 ,年龄 3 4～ 4 5岁 ,月经规律、术前 3个月未用激素 ,其中增殖期 ,分泌早、中、晚期子宫内膜 (病理检查证实 )各 10例。 ( 2 )早孕蜕膜 :选同期在我中心门诊行人工流产术…     

子宫内膜增生组织中白血病抑制因子和巨噬细胞移动抑制因子的检测及临床意义

目的探讨白血病抑制因子（LIF）和巨噬细胞移动抑制因子（MIF）与子宫内膜增生症的关系。方法2005年5月至2006年4月在哈尔滨医科大学附属第四医院和黑龙江省电力医院对85例子宫内膜增生症患者的子宫内膜组织进行分类,并应用免疫组化方法,对组织进行LIF和MIF蛋白检测。结果子宫内膜增生症患者的内膜组织中,LIF检测总阳性率为64.7%,在单纯型、复合型、不典型增生型组织中的阳性率分别为62.7%、61.1%、75.0%。MIF检测总阳性率为72.9%,在各型组织中的阳性率分别为62.7%、83.3%、93.8%。MIF在子宫内膜不典型增生型组织中阳性率与另两型比较差异有显著性意义（P〈0.05）。而LIF在3种类型的子宫内膜增生组织中阳性率差异无显著性意义（P〉0.05）,且两种因子在组织中的表达无明显相关性。结论MIF在子宫内膜不典型增生型组织中阳性率较高,推测其与子宫内膜的异常过度增生和癌变有一定关系。     

PCOS患者子宫内膜白血病抑制因子的表达及意义

目的:探讨多囊卵巢综合征(PCOS)患者不同组织学分期的子宫内膜白血病抑制因子(LIF)的表达与子宫内膜容受性的关系。方法:20例已婚PCOS患者(PCOS组)和26例已婚不孕症患者(对照组)在月经周期第5￣14日及排卵后7￣8 d采集其子宫内膜标本,经HE染色判定内膜组织学分期,采用免疫组织化学法检测内膜LIF的表达。结果:PCOS组间质反应不良和分泌反应欠佳的发生率均高于对照组,差异有统计学意义(χ2=5.000,P<0.05;χ2=7.219,P<0.01)。LIF的表达以腺上皮细胞的胞质为主,其在分泌中期子宫内膜的表达水平均高于增生期(P<0.01)。LIF在PCOS组增生期和分泌中期子宫内膜的表达水平均低于其相应的对照组(P<0.01)。结论:PCOS患者着床窗口期LIF的低表达可能影响着床的多个环节,参与了PCOS患者内分泌紊乱纠正后仍出现妊娠率低、流产率高的发生、发展过程。     

中药红花浸液对植入期子宫内膜白血病抑制因子表达水平的影响

目的:探讨中药红花抗早孕过程中的生物学作用机理。方法:随机将受精后第2天的孕鼠分为3个实验组和对照组,分别经口给予不同浓度的红花液和生理盐水,连续3天,停药后24天取子宫,石蜡包埋切片,进行HE和免疫组化染色,将染色结果输入图像分析仪,观察子宫内膜的形态和白血病抑制因子(leukemia inhibitory factor,LIF)蛋白的表达,并作定量分析。结果:实验组子宫内膜形态有异常改变;免疫组化染色表明,随着药物剂量的增多,子宫内膜细胞LIF蛋白的表达逐渐减少;但对照组呈强阳性表达。结论:中药红花作用于孕鼠子宫内膜细胞,抑制LIF蛋白的表达,从而影响胚胎着床,起到了抗早孕的作用。     

肿瘤转移抑制因子CD82/KAI1在假孕小鼠子宫的表达及其对小鼠胚胎着床的影响

目的:研究CD82/KAI1mRNA及蛋白在假孕d1-8小鼠子宫中的表达规律,以及CD82/KAI1抗体对小鼠胚胎体外发育及着床的影响。方法:用RT-PCR及免疫组织化学技术观察CD82/KAI1mRNA及蛋白在假孕小鼠子宫的表达规律。将8-细胞小鼠胚胎培养于含不同浓度CD82/KAI1抗体的培养液中,观察囊胚形成及脱透明带的情况。妊娠d4小鼠宫角注射CD82/KAI1抗体,于妊娠d8观察小鼠胚胎植入数量。结果:①CD82/KAI1mRNA在假孕d1-8小鼠子宫中均有表达,于d4、d5表达最丰富。CD82/KAI1蛋白在假孕d1-8的子宫基质细胞及腺上皮细胞均有表达,假孕d1-4腔上皮细胞无表达,假孕d5子宫腔上皮细胞出现弱阳性表达,从d6开始子宫腔上皮细胞表达逐渐增强。②一定浓度的CD82/KAI1抗体可明显抑制囊胚的形成率(1∶400,P<0.05)和脱带率(1∶800,P<0.05)。③宫角注射一定量的CD82抗体可以显著提高小鼠胚胎的着床数(8.35±0.17vs4.52±0.24,P<0.05)。结论:CD82/KAI1在妊娠小鼠子宫的表达是非胚胎依赖性的。CD82/KAI1在小鼠胚胎发育和着床过程中发挥重要作用。     

Modulation of leukemia inhibitory factor gene expression and protein biosynthesis in the human fallopian tube

OBJECTIVE: The fallopian tube is the site of fertilization and early embryonic growth and a common site of ectopic implantation. Although the factors responsible for early embryogenesis and implantation are incompletely understood, leukemia inhibitory factor may have an important role in early embryonic development and implantation. We set out to evaluate the production and modulation of leukemia inhibitory factor in the fallopian tube. STUDY DESIGN: We first investigated leukemia inhibitory factor messenger ribonucleic acid levels in fallopian tubes. We then investigated leukemia inhibitory factor messenger ribonucleic acid and protein production in tubal epithelial and stromal cell cultures. RESULTS: Leukemia inhibitory factor messenger ribonucleic acid is expressed in the fallopian tube with only slight variation during the menstrual cycle; however, it is markedly elevated in association with ectopic pregnancy. The level is higher in the tubal mucosa than in the remaining layers and is higher in the more distal segments of the fallopian tube. Estradiol and progesterone did not modulate leukemia inhibitory factor expression in epithelial or stromal cell cultures. Interleukin-1α, tumor necrosis factor-α, and transforming growth factor-β enhanced leukemia inhibitory factor expression in epithelial and stromal cells, with transforming growth factor-β1 enhancing expression by fourfold in stromal cells. Epithelial cells secreted high levels of leukemia inhibitory factor compared with stromal cells (332 ± 89 vs 25 ± 42 pg/mg total protein). Yet stromal cells treated with transforming growth factor-β alone or in combination with epidermal growth factor and platelet-derived growth factor, as well as TNF-α alone or in combination with interleukin-1α enhanced secretion of leukemia inhibitory factor at or above the levels found with epithelial cells. CONCLUSIONS: We speculate that the high constitutive levels of leukemia inhibitory factor expressed in the ampullary portion of the fallopian tube may play a role in early embryonic development. Additionally, elevated expression with ectopic implantation and the marked induction of secretion in the tubal stroma by growth factors and cytokines suggest a link between inflammation, leukemia inhibitory factor, and tubal ectopic pregnancies. (Am J Obstet Gynecol 1996;175:1611-9.)     

LIF及其抗体对小鼠胚胎着床影响的体外研究

目的 :研究外源性白血病抑制因子 (LIF)及其抗体对小鼠胚胎着床的影响。方法 :将妊娠 4d的小鼠胚胎种植于已建立的子宫内膜体外模型上 ,观察不同浓度的LIF及其抗体对小鼠胚胎的粘附、植入及扩展情况。并用RT PCR法测定子宫内膜上皮细胞整合素β3的变化。结果 :不同浓度的LIF促进了胚胎的粘附 (P <0 .0 5 ) ,胚胎的扩展率降低。加入LIF抗体 (5 ,1 0 μg/ml)降低了胚胎的粘附率 (P <0 .0 5 ) ,同时LIF对整合素β3的表达有明显的促进作用 (P <0 .0 1 ) ,加入抗体后整合素β3的表达降低。结论 :LIF可能通过上调整合素β3的表达促进胚胎的粘附和植入     

骨桥蛋白在人子宫内膜、早孕蜕膜和绒毛中的表达及意义

目的:通过检测育龄妇女正常月经周期中不同时期的子宫内膜组织、早孕蜕膜和绒毛组织中骨桥蛋白(osteopontin,OPN)的表达,探讨OPN在子宫内膜表达的变化规律及其在胚胎着床过程中的作用。方法:采用免疫组化和逆转录聚合酶链反应等技术,检测正常月经周期的子宫内膜组织、早孕蜕膜、绒毛中骨桥蛋白的表达。结果:OPN在月经周期的各期子宫内膜均有表达,以分泌中期子宫内膜组织的表达最强,且分泌中、晚期的表达量明显高于增生期和分泌早期(P<0.05)。早孕蜕膜和绒毛组织中OPN均呈强表达,明显高于非孕各期子宫内膜组织(P<0.05)。结论:OPN mRNA及其蛋白在分泌中期(种植窗期)子宫内膜的表达明显升高,推测OPN可能与种植窗的开放密切相关。OPNmRNA及其蛋白在早孕蜕膜和绒毛组织中的表达水平进一步升高,提示OPN在胚胎着床后也有重要作用。     

The role of leukemia inhibitory factor in tubal ectopic pregnancy

Introduction

Ectopic pregnancy is unique to humans and a leading cause of maternal morbidity and mortality. The etiology remains unknown however factors regulating embryo implantation likely contribute. Leukemia inhibitory factor (LIF) has roles in extravillous trophoblast adhesion and invasion and is present in ectopic implantation sites. We hypothesised that LIF facilitates blastocyst adhesion/invasion in the Fallopian tube, contributing to ectopic pregnancy.

Methods

We immunolocalised LIF receptor (R) in tubal ectopic pregnancy (N = 5). We used an oviduct cell line (OE-E6/E7) to model Fallopian tube epithelial cells and a trophoblast spheroid co-culture model (HTR-8/SVneo cell line formed spheroids) to model blastocyst attachment to the Fallopian tube. We examined LIF signaling pathways in OE-E6/E7 cells by Western blot. The effect of LIF and LIF inhibition (using a novel LIF inhibitor, PEGLA) on first-trimester placental outgrowth was determined.

Results

LIFR localised to villous and extravillous trophoblast and Fallopian tube epithelium in ectopic pregnancy. LIF activated STAT3 but not the ERK pathway in OE-E6/E7 cells. LIF stimulated HTR-8/SVneo spheroid adhesion to OE-E6/E7 cells which was significantly reduced after PEGLA treatment. LIF promoted placental explants outgrowth, while co-treatment with PEGLA blocked outgrowth.

Discussion

Our data suggests LIF facilitates the development of ectopic pregnancy by stimulating blastocyst adhesion and trophoblast outgrowth from placental explants. Ectopic pregnancy is usually diagnosed after 6 weeks of pregnancy, therefore PEGLA may be useful in targeting trophoblast growth/invasion.

Conclusion

LIF may contribute to the development of ectopic pregnancies and that pharmacologically targeting LIF-mediated trophoblast outgrowth may be useful as a treatment for ectopic pregnancy.     

Leukemia inhibitory factor in human reproduction

Objective: To describe the clinical findings, expressions, interactions, and clinical implications of leukemia inhibitory factor (LIF) in human reproduction.

Design: Review of published articles.

Setting: Clinical development unit of biotechnology company.

Intervention(s): None.

Result(s): In the endometrium, LIF is expressed in a menstrual cycle–dependent manner, with the highest level occurring at the time of implantation. LIF is also detected in uterine flushing, and its level is significantly lower in women with unexplained infertility. Likewise, endometrial explants derived from women with unexplained infertility showed reduced levels of LIF secretion. Binding of LIF to LIF receptor and gp130 activates signal transduction pathways. LIF receptor is expressed in endometrium, oocytes, and blastocysts. Cytotrophoblasts cultured in the presence of LIF differentiate toward an anchoring extravillous phenotype.

Conclusion(s): On the basis of reports gathered from animal and human studies, LIF appears to play an important role in implantation and in the establishment of pregnancy.