温肾活血方改善胚胎着床障碍小鼠子宫内膜容受性的研究

目的:探讨温肾活血方对胚胎着床障碍模型小鼠妊娠结局及子宫内膜容受性的影响。方法:由米非司酮建立胚胎着床障碍模型小鼠200只,随机分为正常(N)组、模型(M)组、中药低剂量(L)组、中药高剂量(H)组和孕酮(P)组各40只。妊娠第1~4日早晨,H组、L组小鼠灌胃给予0.3 m L水煎剂,N组、M组小鼠每日以等体积生理盐水灌胃,P组小鼠以配制的黄体酮-橄榄油剂按照2 mg/只进行颈后皮下注射,妊娠第4日上午9︰00造模,N组小鼠颈后皮下注射0.1 m L橄榄油溶剂,其他各组小鼠颈后皮下注射米非司酮-橄榄油剂0.1 m L(1.93 mg/kg米非司酮)。分别于妊娠第4日、第5日、第8日上午9∶00脱颈椎处死每组各10只小鼠,检测妊娠率、着床位点数、子宫内膜组织形态、雌激素(E_2)及其受体(ER)、孕激素(P)及其受体(PR)、胞饮突。结果:N组着床位点数多于其它各组,H组与N组比较无统计学差异(P0.05)。子宫内膜发育水平N组最优,H组接近N组,M组滞后。妊娠第4日,N组的E_2、P和ER、PR的表达水平与M组比较无统计学差异(P0.05),低于P组与H组(P0.01);妊娠第5日,N组的E_2、P和ER、PR的表达水平显著高于M组(P0.05),N组的ER水平与P、H、L三组比较无统计学差异(P0.05),N组的PR高于P组和L组(P0.05),与H组比较均无统计学差异(P0.05)。P组P水平高于其它组(P0.01),E_2水平与ER结果相吻合。M组胞饮突发育滞后,H组胞饮突发育程度接近N组,P组偶见退化期胞饮突。结论:温肾活血方能够改善子宫内膜容受性,且其效果优于黄体酮。     

胞饮突与子宫内膜容受性

胞饮突是在扫描电镜下所见宫腔被复上皮细胞膜预端出现的膜突起。在自然周期,它成熟的时间平均在月经周期的第２１天。激素治疗可使胞饮突的出现提前或延迟,在卵巢刺激周期,胞饮突比自然周期提前１～２天出现,相反,用雌激素和孕激素行人工周期治疗,胞饮突的形成推迟。由于它的出现不仅与鼠和人类子宫内膜容受胚胎着床的时间一致。而且有可能参与囊胚的着床过程,因此被认为是子宫内膜受性或着床窗的特异形态标记。     

对胚泡着床障碍小鼠子宫内膜容受性的研究

目的:建立胚泡着床障碍小鼠模型,并研究其子宫内膜容受性。方法:于妊娠d4上午利用米非司酮诱导昆明小鼠胚泡着床障碍,12h后处死小鼠,观察小鼠胚泡着床率及平均着床胚泡数,利用光镜观察小鼠子宫内膜形态结构,采用免疫组化SP法检测子宫雌、孕激素受体(ER、PR)表达,采用RT-PCR检测子宫ER、PR基因表达。结果:与对照组相比,模型组小鼠胚泡着床率及平均着床胚泡数明显降低,子宫内膜发育明显受抑制;子宫内膜腺体、间质ER、PR表达范围和强度均显著降低。结论:米非司酮能成功诱导建立胚泡着床障碍模型,其着床期子宫内膜发育受抑制,子宫内膜容受性降低,胚泡着床失败。     

Meis1在围着床期小鼠子宫内膜中的表达

目的:探讨Meis1在围着床期小鼠子宫内膜中的作用。方法:采用免疫组织化学定位检测Meis1在小鼠子宫内膜中的表达;采用Real-time RT-PCR和免疫印迹方法,分别在mRNA和蛋白水平定量检测Meis1在围着床期小鼠子宫内膜中的表达水平。结果:免疫组织化学结果显示,Meis1蛋白主要表达于小鼠子宫内膜腺上皮细胞的细胞膜和胞质以及基质细胞和蜕膜细胞的细胞核中,在围着床期小鼠子宫内膜中的表达随妊娠天数的增加而增加,着床后在基质和蜕膜中的表达增强;在孕第4日显著增加,孕第5日达高峰,孕第6日表达下降。Real-time RT-PCR和免疫印迹结果显示,Meis1的表达随妊娠天数的增加逐渐增强,孕第4日明显增强,孕第5日达高峰,孕第6日开始下降。结论:Meis1在围着床期小鼠子宫内膜中呈规律表达,与小鼠着床窗吻合,提示Meis1是调控围着床期子宫内膜容受性的重要因子。     

Asoprisnil抗小鼠孕卵着床效果及对子宫内膜容受性的影响

目的:探讨Asoprisnil抗小鼠孕卵着床的效果及其对小鼠着床窗期子宫内膜容受性的影响。方法:将孕第1日小鼠随机分成4组,分别为Asoprisnil低(5μg/g)、中(10μg/g)、高(20μg/g)剂量组和对照组,每组22只,于孕第1~3日每日灌胃给药1次,对照组以体积分数1%羧甲基纤维素钠替代,第5日每组处死孕鼠5只取子宫组织,HE染色观察子宫内膜形态学改变,免疫组织化学S-P法检测子宫内膜PCNA表达。第8日处死余下的孕鼠,计数着床点数。结果:①Asoprisnil高、中和低剂量组的妊娠率分别为11.76%、35.29%和76.47%,与对照组(94.12%)比较差异有统计意义(P<0.001)。②Asoprisnil高、中、低剂量组着床胚泡数第50百分位数(P50)分别为13(10.5~14)、10(0.5~11)和0(0~12),与对照组0(0~0)比较差异均有统计学意义(P<0.001)。③对照组围着床期子宫内膜腺体弯曲折叠,为复层高柱状上皮细胞;基质细胞蜕膜变,细胞大且排列疏松,胞质丰富透亮;内膜中腺体和血管丰富。Asoprisnil组子宫内膜腺体为单层或者复层上皮细胞;内膜基质细胞蜕膜变不明显,基质细胞较小,排列致密;腺体和血管增生不明显。④围着床期小鼠子宫内膜腺体和基质中均有PCNA表达。内膜腺体中Asoprisnil高、中、低剂量组PCNA表达强度(AIOD)分别为0.15±0.01、0.16±0.03和0.14±0.02,与对照组(0.21±0.03)比较差异有统计学意义(P<0.001)。Asoprisnil高、中、低剂量组内膜基质细胞中PCNA表达强度(AIOD)分别为0.17±0.01、0.18±0.03和0.17±0.02,与对照组(0.15±0.02)比差异有统计学意义(P<0.001)。结论:Asoprisnil能显著抑制着床窗期子宫内膜腺体增殖,促进子宫内膜基质细胞增殖,但阻止基质细胞蜕膜化,降低子宫内膜容受性而发挥抗小鼠胚胎着床效应,显示出潜在的子宫内膜靶向避孕前景。     

补肾益气和血方中药对胚泡着床障碍小鼠子宫内膜表面胞饮突表达的影响

目的　观察补肾益气和血方中药对胚泡着床障碍小鼠子宫内膜表面胞饮突表达的影响。方法　选择 8～ 12周龄昆明小鼠 ,于妊娠第 4天 (Pd4 )晨 ,皮下注射米非司酮 ,制成胚泡着床障碍模型。然后将妊娠小鼠随机分为正常组、模型组和治疗组 ,治疗组从Pd1开始灌服中药 (补肾益气和血方 ) ,正常组和模型组则给予等量生理盐水灌服 ,模型组和治疗组于Pd4晨皮下注射米非司酮 ,正常组则注射等量生理盐水。采用扫描电镜观察各组小鼠在Pd4晚 2 1时 30分～ 2 2时 (第 1时间点 ,T1)和Pd5上午 9时 30分～ 10时 (第 2时间点 ,T2 ) ,共两个时间点子宫内膜表面胞饮突的表达。结果　模型组妊娠率 ( 2 5 % )较正常组 ( 90 % )显著降低 ,差异有显著性 (P <0 0 5 ) ,模型组平均着床胚泡数为 ( 7 7± 1 3)个 ,正常组为 ( 14 7± 1 4 )个 ,两组比较 ,差异有极显著性 (P <0 0 0 1) ;治疗组的妊娠率 ( 5 5 % )及平均着床胚泡数 [( 12 3± 0 8)个 ]均较模型组显著增加 (P <0 0 5 ,P <0 0 1)。T1时正常组子宫内膜表面表达大量发育中的胞饮突 ,至T2时则表达大量发育完全的胞饮突 ;T1时模型组子宫内膜发育不同步 ,胞饮突仅在局部少量表达 ,至T2时则表达完全消失 ;T1时治疗组子宫内膜表面表达大量发育中的胞饮突 ,与正常组相比略延     

子宫内膜异位症与胚胎着床

胚胎着床的过程极为复杂,包括定位、粘附和侵入3个阶段.着床仅发生在一个极其有限的时间与空间范围内(即所谓的"着床窗"),需要子宫内膜和胚胎发育的同步化、相互作用和配合.目前,胚胎着床机制尚未充分阐明,但胚胎成功着床的要素包括:高质量的胚胎、有接受性的子宫内膜和适宜的内分泌环境.     

子宫内膜容受性的调控及其特征标记的研究进展

胚泡着床是一复杂的生理过程.着床只发生在具有接受性的子宫内膜,子宫对胚泡的接受性叫容受性.子宫内膜容受性的调控机制尚不清楚,涉及卵巢甾体激素,细胞因子和黏附分子等的自分泌及旁分泌机制.综述子宫内膜容受性的调节机制及特征性标记.     

子宫内膜容受性的调控及其特征标记的研究进展

胚泡着床是一复杂的生理过程。着床只发生在具有接受性的子宫内膜,子宫对胚泡的接受性叫容受性。子宫内膜容受性的调控机制尚不清楚,涉及卵巢甾体激素,细胞因子和黏附分子等的自分泌及旁分泌机制。综述子宫内膜容受性的调节机制及特征性标记。     

子宫内膜容受性的评价及其改善措施

子宫内膜的容受性是指子宫内膜对胚胎的接受能力。部分不孕症患者的子宫内膜容受性存在缺陷,影响了胚胎的成功着床。因此,如何对其做出合理的评价和相应的改善措施以提高妊娠率,是世界生殖医学系统研究的焦点。本文分别从形态学、超声学和分子生物学三个角度对子宫内膜容受性做出了评价。激素调节、抗凝剂的使用和中医中药等药物干预方法可有效改善子宫内膜容受性。此外,机械刺激和对输卵管积水的患者行患侧输卵管切除术对提高患者的子宫内膜容受性也起到了积极的作用。     

An animal model of effects of nicotine exposure on endometrial receptivity and embryo implantation in pregnancy

Objective: This study aims at evaluating the endometrial receptivity in uterus of pregnant rats exposed to nicotine via examination of integrin expression by immunohistochemical effect.

Methods: In this study, 16 healthy pregnant rats were divided into two groups of control and study groups each comprising eight rats. The rats randomised to study group were given a certain amount of nicotine before and during the pregnancy. Integrin expression was detected in uterus of all rats by immunohistochemical staining. The effect of nicotine exposure on embryo implantation and the endometrial receptivity were immunohistochemically and pathologically evaluated.

Results: Comparison of both groups revealed no difference in living, viable foetuses. Intensity and universality of immunohistochemical staining of Integrin β3 for endometrial epithelium and endometrial stroma were detected to be identical between the groups.

Conclusion: No immunochemical effect was observed on integrin expression, which is a very important part of receptivity in an animal model created with pregnant rats that were transdermally exposed to nicotine. Our study demonstrated that the harmful effect of nicotine use before and pregnancy on implantation is limited at the level of integrin expression, in a dose-dependent manner and also by considering the method of administration.     

胚胎着床研究进展

胚胎着床过程包括定位、黏附、侵入,其调控机制复杂而精细,涉及多个基因、生物分子、细胞因子和信号传导通路,其中胚胎质量、子宫内膜容受性、胚胎发育与子宫内膜发育的同步性,以及良好的母胎对话是胚胎着床所必需。胚胎着床的基础和临床研究取得的进展,有助于进一步探索胚胎着床的奥秘,加深了解胚胎着床的相关机制,对不孕症的诊治具有重要意义。     

Ultrastructural markers of tissue endometrial receptivity in patients with recurrent implantation failure

Abstract

The scanning electron microscopy of the endometrial surface epithelium during the ‘implantation window’ was performed in 119 patients with uterine factor of infertility or recurrent miscarriage due to endometrial hypoplasia. Ultramorphological picture of the surface endometrial epithelium was characterized by aplasia and hypoplasia of pinopodes (67.39%), dense cell – cell contacts (69.53%), heteromorphy of secretory cells (15.22%) in combination with atypia of microenvironment cells (50%) in patients with infertility. The asynchronous development of pinopodes (46.67%) and the absence of intercellular contacts separation during the ‘implantation window’ (84.44%) was observed in patients with recurrent miscarriage. The revealed disturbance determines the mechanisms of the blastocyst adhesion violation and trophoblast invasion in the different stages of implantation in patients with uterine factor of infertility and recurrent miscarriage.     

Clinical utility of the endometrial receptivity analysis in women with prior failed transfers

PurposeTo determine the utility of the endometrial receptivity analysis (ERA) in women with prior failed embryo transfers (ET).MethodsThis was a retrospective study of patients who underwent an ERA test with a subsequent frozen ET. Women were classified based on their indication for an ERA test: (1) ≥ 1 prior failed ET (cases), or (2) as a prophylactic measure (controls). A subset analysis of women with ≥ 3 prior failed transfers was performed. Pregnancy outcomes of the subsequent cycle were examined, including conception, clinical pregnancy, and ongoing pregnancy/live birth.ResultsA total of 222 women were included, 131 (59%) women with ≥ 1 prior failed ET and 91 (41%) controls. Among the 131 women with ≥ 1 prior failed ET, 20 women (9%) had ≥ 3 prior failed ETs. The proportion of non-receptive ERA tests in the three groups were the following: 45% (≥ 1 prior failed ET), 40% (≥ 3 prior failed ETs), and 52% (controls). The results did not differ between cases and controls. The pregnancy outcomes did not differ between women with ≥ 1 prior failed ET and controls. In women with ≥ 3 prior failed ETs, there was a lower ongoing pregnancy/live birth rate (28% vs 54%, P = 0.046).ConclusionWomen with ≥ 1 prior failed ET and ≥ 3 prior failed ETs had a similar prevalence of non-receptive endometrium compared to controls. Women with ≥ 3 prior failed ETs had a lower ongoing pregnancy/live birth rate despite a personalized FET, suggesting that there are additional factors in implantation failure beyond an adjustment in progesterone exposure.     

降调节联合激素替代冻融胚胎移植周期中不同内膜转化时间对反复种植失败患者临床结局的影响

目的:探讨反复种植失败(RIF)患者冻融胚胎移植(FET)周期移植时机的选择及对临床结局的影响。方法:选择采用Gn RH-a降调节激素替代方案准备子宫内膜进行FET的RIF患者106例,随机分为A组(常规法)53个周期,即孕酮作用内膜3 d后移植第3日胚胎;B组(改良法)53个周期,即延迟内膜扳机和黄体支持用药,比较组间血清性激素水平、临床妊娠率、早期流产率等相关指标。结果:患者的基本情况组间具有可比性(P0.05);移植日雌二醇(E_2)、孕酮(P)、E_2/P、临床妊娠率组间均有统计学差异(P0.05);移植前1日血清P水平组间差异有统计学意义(P0.05)。结论:对于RIF患者,在降调节联合激素替代-FET周期中,延长雌、孕激素用药时间可刺激内膜达到理想状态,进而获得满意的妊娠结局。     

宫腔粘连模型大鼠子宫内膜胞饮突发育和整合素β3表达

目的:建立宫腔粘连大鼠模型,观察子宫内膜胞饮突的发育和整合素β3的表达,探讨宫腔粘连对子宫内膜容受性的影响。方法:选取健康SD大鼠30只,将每只大鼠左侧子宫作为模型组(宫腔注入95%无水乙醇,持续5min),右侧子宫作为对照组(注入等量生理盐水)。术后两个动情周期取子宫,光镜观察子宫内膜形态学变化及纤维化程度,扫描电镜观察大鼠子宫内膜胞饮突的发育情况,免疫组化法检测子宫内膜整合素β3表达。结果:模型组大鼠子宫内膜明显变薄,上皮细胞排列紊乱,间质内胶原纤维增多,子宫内膜纤维化评分为(4.69±0.22)分,明显高于对照组。扫描电镜观察,对照组大鼠子宫内膜可见大量发育完全的胞饮突,而模型组子宫内膜胞饮突分布稀少,表面皱缩,发育不同步。模型组的整合素β3表达明显低于对照组,差异有统计学意义(P0.05)。结论:注射无水乙醇可建立稳定的大鼠宫腔粘连动物模型,抑制子宫内膜胞饮突和整合素β3表达,降低子宫内膜的容受性。