NGF 及其受体TrkA 在子宫腺肌病中的表达及与痛经的关系

目的:研究NGF及其受体TrkA在子宫腺肌病患者的异位内膜与在位内膜组织的表达情况及与痛经的关系。方法:采用免疫组化MaxVision法检测子宫腺肌病异位内膜(30例)、在位内膜(30例)、正常子宫内膜(19例)标本中NGF、TrkA蛋白的表达,分析其表达差异及与痛经的关系。结果:①子宫腺肌病异位内膜组NGF、TrkA表达显著高于正常内膜组(P<0.01),在位内膜组NGF、TrkA表达显著高于正常内膜组(P<0.01),子宫腺肌病异位内膜组NGF、TrkA表达与在位内膜组无显著差异。②子宫腺肌病异位内膜组NGF、TrkA表达与痛经强度评分呈正相关(相关系数r=0.637,P=0.000;r=0.662,P=0.000)。结论:NGF及其受体TrkA在子宫腺肌病中高表达可能参与子宫腺肌病发病机制,而且可能与痛经有关。     

NGF及其受体TrkA在子宫腺肌病中的表达及与痛经的关系

目的：研究NGF及其受体TrkA在子宫腺肌病患者的异位内膜与在位内膜组织的表达情况及与痛经的关系。方法：采用免疫组化MaxVision法检测子宫腺肌病异位内膜（30例）、在位内膜（30例）、正常子宫内膜（19例）标本中NGF、TrkA蛋白的表达,分析其表达差异及与痛经的关系。结果：①子宫腺肌病异位内膜组NGF、TrkA表达显著高于正常内膜组（P〈0.01）,在位内膜组NGF、TrkA表达显著高于正常内膜组（P〈0.01）,子宫腺肌病异位内膜组NGF、TrkA表达与在位内膜组无显著差异。②子宫腺肌病异位内膜组NGF、TrkA表达与痛经强度评分呈正相关（相关系数r=0.637,P=0.000;r=0.662,P=0.000）。结论：NGF及其受体TrkA在子宫腺肌病中高表达可能参与子宫腺肌病发病机制,而且可能与痛经有关。     

神经细胞黏附因子在子宫腺肌病病灶中的表达及意义

目的:探究神经细胞黏附因子(NCAM)在子宫腺肌病病灶中的表达及意义。方法:将2016年6月~2017年6月在我院接受治疗的80例子宫腺肌病患者作为研究对象,包括分泌期与增生期各40例,采用免疫组化法检测,其在位内膜、异位内膜组织中NCAM表达情况,并以同期我院40例正常者子宫内膜标本作为对照。对子宫腺肌病患者痛经程度给予NRS疼痛评估,比较其NCAM的表达。结果:80例子宫腺肌病在位内膜、异位内膜均存在NCAM表达,40例正常内膜腺上皮有38例存在NCAM表达,2例正常内膜无表达。NCAM在异位内膜组织中的表达明显高于在位内膜及正常子宫内膜(P0.05),差异均有统计学意义。NCAM在在位内膜组织分泌期表达与增生期差异有统计学意义(P0.05);子宫腺肌病异位病灶NCAM表达与患者NRS评分呈现明显正相关(r=0.824,P0.05)。结论:NCAM在异位子宫内膜高表达,可能参与了子宫内膜异位症的发生和发展,并与患者痛经程度呈现出正相关。     

子宫腺肌病中异位和正位子宫内膜血管发生的研究

目的探讨子宫腺肌病中异位和正位内膜组织中血管内皮生长因子(VEGF)的表达及微血管密度(MVD)与发病机制的关系.方法采用免疫组织化学技术检测21例正常子宫内膜(对照组)和38例子宫腺肌病(疾病组)异位内膜及正位内膜中VEGF的表达,利用Ⅷ因子相关抗原标记血管内皮细胞,再应用计算机图像分析系统分析VEGF免疫染色密度值及计数切片中微血管(20×视野).结果疾病组异位内膜腺上皮细胞及间质细胞中VEGF的表达明显高于对照组腺上皮及间质细胞(P<0.01).疾病组正位内膜腺上皮细胞中的VEGF的表达亦高于对照组(P<0.05).疾病组异位内膜中的平均微血管密度明显高于正位内膜及对照组(P<0.05).VEGF与MVD表达呈明显正相关(P<0.01).结论子宫腺肌病异位和正位内膜中VEGF表达的明显增高及局部微血管密度的明显增加,可能导致局部新生毛细血管的增多,从而促进子宫内膜在子宫肌层内异位种植和生长.     

基质金属蛋白酶-2,-9在子宫腺肌病的表达

目的　探讨基质金属蛋白酶 2 , 9(MMP 2 , 9)在子宫腺肌病的表达及与发病机制的关系。方法　采用免疫组织化学SP法对 2 8例子宫腺肌病 (疾病组 )异位及原位内膜 ,35例子宫肌瘤 (对照组 )原位内膜中MMP 2 , 9的表达进行测定 ,应用图像分析系统分析MMP 2 , 9平均光密度值。结果　MMP 2 , 9于两组原位内膜中表达均呈月经周期依赖性 ,分泌期的表达强度高于增生期 (P <0 0 5 ) ;于疾病组异位内膜中表达无周期性显著差异 (P >0 0 5 )。疾病组内增生期异位内膜MMP 2 , 9的表达强度高于原位内膜 (P <0 0 5 )。疾病组整个月经周期中 ,MMP 2 , 9于原位及异位内膜的表达皆持续明显高于对照组 (P <0 0 5 )。结论　研究显示子宫腺肌病原位和异位内膜MMP 2 , 9的过度表达可能与其发病机制有关     

基质金属蛋白酶MMP-2及其抑制物TIMP-2在子宫腺肌病中的表达

目的:探讨基质金属蛋白酶MMP-2及其抑制物TIMP-2在子宫腺肌病异位内膜细胞及在位内膜细胞中的表达.方法:应用免疫组化方法(SP法)检测MMP-2及其抑制物TIMP-2在46例子宫腺肌病异住内膜细胞,在位内膜细胞及30例子宫肌瘤内膜细胞的表达.结果:MMP-2蛋白在子宫肌瘤内膜细胞组中阳性表达率为33.33%,在子宫腺肌病异位内膜细胞组中阳性率为89.13%,在位内膜细胞中阳性率为84.78%,差异具有显著性(P＜0.01).TIMP-2蛋白在子宫肌瘤内膜细胞组中阳性表达率为46.67%,在子宫腺肌病异位内膜细胞组中阳性率为15.22%,在位内膜细胞中为47.83%,差异有显著性(P＜0.01).结论:子宫腺肌病患者异位内膜组织中MMP-2表达增高,TIMP-2表达降低,使MMP-2与TIMP-2平衡失调,从而参与了子宫腺肌病的发生发展.     

HLA-G蛋白及基因在子宫腺肌病中的表达

目的:探讨人类白细胞抗原G(human leucocyte antigen-G,HLA-G)在子宫腺肌病(adenomyosis,AM)患者在位、异位内膜中的表达及其生物意义,为AM的病因研究和治疗展望提供依据.方法:采用免疫组化和RT-PCR法检测36例AM患者和30例正常子宫内膜在位、异位内膜HLA-G蛋白及基因表达情况.结果:AM在位、异位内膜HLA-G蛋白及基因表达量显著高于正常子宫内膜(P<0.05);AM在位与异住内膜HLA-G蛋白表达量无显著差异(P>0.05);AM在位、异位内膜腺体细胞HLA-G蛋白的表达量呈正相关(r=0.948,P<0.05);AM在位、异位子宫内膜间质细胞HLA-G蛋白的表达量也呈正相关(r=0.863,P<0.05).结论:HLA-G在AM在位、异位内膜中呈高表达,参与了AM的发病机制,HLA-G阳性的子宫内膜更容易逃避免疫细胞的攻击而浸润肌层生长.     

子宫内膜干细胞及其与血管生成关系的研究进展

大量的临床观察和基础研究均提示,子宫内膜中存在一些干细胞,它们具有高度增殖、自我更新和的分化潜能。目前有学者研究子宫内膜干细胞是否可能成为子宫内膜病变患者进行内膜修复替代治疗的手段,以及是否可以在毛细血管重建及血管发生中发挥作用。虽然近年来在子宫内膜干细胞的鉴定和分离方面有了很大的进展,但对于它的特异性标志物仍在探索中。同时对子宫内膜干细胞的研究可为内膜异位症、子宫内膜癌等疾病的治疗提供新的思路。本文针对国内外研究者近年来在这发面的研究现状进行综述。     

Survivin和MMP-9在子宫上皮和间质细胞中的表达与子宫内膜异位症间的关系

目的研究Survivin蛋白和基质金属蛋白酶9（matrix metalloproteinase-9,MMP-9）在子宫腺肌症（ade-nomyosis,AM）、腹壁子宫内膜异位症（abdominal wall endometriosis,AWEMS）、卵巢子宫内膜异位症（ovary endometri-osis,OEMS）的在位、异位内膜的腺上皮细胞和间质细胞中的表达,并与对照组进行比较,探讨其在子宫内膜异位症（endometriosis,EMS）的发生发展中的作用。方法采用免疫组化法检测各组织标本中Survivin和MMP-9的表达。结果 （1）在正常子宫内膜组织中Survivin、MMP-9均呈弱表达或无表达。在EMS三个病例组中,无论是在位内膜还是异位内膜组织Survivin、MMP-9的表达均有不同程度的上调,分别与正常子宫内膜组织表达比较差异有统计学意义（P〈0.05）。（2）在AM、AWEMS、OEMS三个病例组中,仅在增生期异位内膜腺上皮细胞和间质细胞中Sur-vivin、MMP-9的表达分别高于在位内膜同类细胞的表达,差异均有统计学意义（P〈0.05）;分泌期则呈不规律表达。（3）在AM、AWEMS、OEMS三个病例组中,限定相同组织部位、相同细胞类型,增生期与分泌期Survivin、MMP-9的表达水平差异无统计学意义（P〉0.05）且无周期性。（4）在AM、AWEMS、OEMS三个病例组中,限定相同生理期、相同组织部位比较腺上皮细胞与间质细胞Survivin、MMP-9的表达：腺上皮细胞显著高于间质细胞的表达,差异有统计学意义（P〈0.05）。（5）EMS三个病例组之间,限定相同生理期、相同组织部位、相同细胞类型,组间分别比较Survivin或MMP-9的表达水平：Survivin表达差异无统计学意义（P〉0.05）,异位内膜仅分泌期MMP-9在AWEMS腺上皮细胞的表达（4.45±0.18）和AM腺上皮细胞的表达（4.68±0.17）高于OEMS异位内膜腺上皮细胞的表达（2.13±0.12）,差异均有统计学意义（P〈0.05）。结论 Survivin和MMP-9在EMS在位内膜和异位内膜腺上皮细胞和间质细胞中高表达可能是内异症发生发展的重要因素并有协同作用,在位内膜异常是EMS发病的决定性因素,腺上皮细胞高表达在EMS的发生发展中起主导作用,AM、AWEMS、OEMS三个病例组中的生物学特性不同可能受发病诱因、腹腔内环境及多种相关因子影响。     

FAS在子宫内膜样腺癌及其癌前病变中的表达及意义

目的研究脂肪酸合成酶(Fatty acid synthase FAS)在子宫内膜样腺癌及其癌前病变中的表达以及与临床病理意义。方法 37例子宫内膜样腺癌、21例子宫内膜非典型增生、23例子宫内膜复杂性增生、17例子宫内膜单纯性增生及11例增生期子宫内膜中FAS的表达情况,结合临床病理参数并进行统计学分析。结果 FAS在子宫内膜样腺癌、子宫内膜非典型增生、子宫内膜复杂性增生、子宫内膜单纯性增生及增生期子宫内膜中均有表达,其阳性表达率依次为81.1%、57.1%、56.5%、52.9%、45.5%。子宫内膜样腺癌中FAS的阳性表达率明显高于增生期子宫内膜、子宫内膜单纯性增生、子宫内膜复杂性增生及子宫内膜非典型增生(P<0.05)。FAS的阳性表达率在浸润深度≥1/2肌层的子宫内膜样腺癌明显高于浸润深度<1/2肌层者(P<0.05)。FAS的表达与年龄及组织学分级无显著差异性(P>0.05)。在子宫内膜样腺癌中FAS的表达与ER有关(P<0.05)结论本研究提示FAS的表达可能与子宫内膜样腺癌的发生、发展有关。FAS可以作为提示子宫内膜样腺癌预后的一个新的参考指标。部分ER阳性的子宫内膜样腺癌的发生、发展可能与FA...     

Proteomics identification of annexin A2 as a key mediator in the metastasis and proangiogenesis of endometrial cells in human adenomyosis

Adenomyosis is a common estrogen-dependent disorder of females characterized by a downward extension of the endometrium into the uterine myometrium and neovascularization in ectopic lesions. It accounts for chronic pelvic pain, dysmenorrhea, menorrhagia, and infertility in 8.8-61.5% women worldwide. However, the molecular mechanisms for adenomyosis development remain poorly elucidated. Here, we utilized a two-dimensional polyacrylamide gel electrophoresis/MS-based proteomics analysis to compare and identify differentially expressed proteins in matched ectopic and eutopic endometrium of adenomyosis patients. A total of 93 significantly altered proteins were identified by tandem MS analysis. Further cluster analysis revealed a group of estrogen-responsive proteins as dysregulated in adenomyosis, among which annexin A2, a member of annexin family proteins, was found up-regulated most significantly in the ectopic endometrium of adenomyosis compared with its eutopic counterpart. Overexpression of ANXA2 was validated in ectopic lesions of human adenomyosis and was found to be tightly correlated with markers of epithelial to mesenchymal transition and dysmenorrhea severity of adenomyosis patients. Functional analysis demonstrated that estrogen could remarkably up-regulate ANXA2 and induce epithelial to mesenchymal transition in an in vitro adenomyosis model. Enforced expression of ANXA2 could mediate phenotypic mesenchymal-like cellular changes, with structural and functional alterations in a β-catenin/T-cell factor (Tcf) signaling-associated manner, which could be reversed by inhibition of ANXA2 expression. We also proved that enforced expression of ANXA2 enhanced the proangiogenic capacity of adenomyotic endometrial cells through HIF-1α/VEGF-A pathway. In vivo, we demonstrated that ANXA2 inhibition abrogated endometrial tissue growth, metastasis, and angiogenesis in an adenomyosis nude mice model and significantly alleviated hyperalgesia. Taken together, our data unraveled a dual role for ANXA2 in the pathogenesis of human adenomyosis through conferring endometrial cells both metastatic potential and proangiogenic capacity, which could serve as a potential therapeutic target for the treatment of adenomyosis patients.     

子宫内膜息肉研究进展

子宫内膜息肉(endometrial polyps,EMP)由子宫内膜腺体和含有厚壁血管的纤维化内膜间质构成,是局部子宫内膜过度增生形成的有蒂或无蒂的赘生物。子宫内膜息肉是最常见的子宫内膜病变之一,临床表现为子宫不规则出血,或月经量增多、不孕、绝经后出血等,也可无明显临床症状。子宫内膜息肉多数属良性病变,但其可恶变性已经被证实。子宫内膜息肉的发病机制目前尚不明确,传统观点认为其与慢性子宫内膜炎症有关,属慢性炎症范畴,即为生物致炎因子及长期反复机械性刺激所致的反应性增生物;近年来随着分子生物学研究的深入,发现子宫内膜息肉的发生与可能与激素调控下增殖与凋亡失衡相关。近来随着宫腔镜检查技术的推广及激素补充治疗人数的增多,子宫内膜息肉的发病率及检出率逐渐增加,加之经刮宫或电切治疗后复发率非常高,因此,子宫内膜息肉越来越受到临床医生的重视,现将子宫内膜息肉的研究进展作一综述。     

Increase in the number of integrinbeta1-immunoreactive monocyte-lineage cells in experimentally-induced adenomyosis in mice

Uterine adenomyosis is a disease in which hyperplastic endometrial stroma and glands invade the myometrium. We have previously demonstrated that hyperprolactinemia leads to the development of adenomyosis in mice. In the present study, a subtracted cDNA library was made by suppression subtractive hybridization to find specific genes that are abundantly expressed in the adenomyotic but not normal tissue in mice. A cDNA fragment of integrinbeta1 (ibeta1) was found in the library, and the expression of the gene product was increased in the adenomyotic uteri at mRNA and protein levels. Intense ibeta1-immunoreactivity was localized on a group of cells dispersing throughout the endometrial stroma. The number of ibeta1-immunoreactive (ibeta1-ir) cells was significantly greater in the uteri of mice with adenomyosis than normal mice. The majority of the ibeta1-ir cells expressed CD14-ir signal, a marker for monocyte-lineage cells, whereas an increase in the number of CD14-ir cells was also evident in the adenomyotic uteri, especially in the ectopic endometrial tissue. Thus, the adenomyotic stromal tissue contained numerous monocyte-lineage cells with higher expression levels of ibeta1, one of their products. The relationship between the increased number of monocyte-lineage cells and the hyperplastic proliferation of endometrial tissues was discussed with a view to understanding the progressive mechanism of adenomyosis.     

A new trick for an old dog: The application of mifepristone in the treatment of adenomyosis

Adenomyosis is also called internal endometriosis and affects about 20% of reproductive‐aged women. It seriously reduces life quality of patients because current drug therapies face with numerous challenges. Long‐term clinical application of mifepristone exhibits wonderful therapeutic effects with mild side‐effects in many disorders since 1982. Since adenomyosis is a refractory disease, we investigate whether mifepristone can be applied in the treatment of adenomyosis. In this study, we investigated the direct effects of mifepristone on human primary eutopic endometrial epithelial cells and stromal cells in adenomyosis. We found that mifepristone causes cell cycle arrest through inhibiting CDK1 and CDK2 expressions and induces cell apoptosis via the mitochondria‐dependent signalling pathway in endometrial epithelial cells and stromal cells of adenomyosis. Furthermore, mifepristone inhibits the migration of endometrial epithelial cells and stromal cells through decreasing CXCR4 expression and restricts the invasion of endometrial epithelial cells via suppression of epithelial‐mesenchymal transition in adenomyosis. We also found that mifepristone treatment decreases the uterine volume, CA125 concentration and increases the haemoglobin concentration in serum for adenomyosis patients. Therefore, we demonstrate that mifepristone could serve as a novel therapeutic drug in the treatment of adenomyosis, and therefore, the old dog can do a new trick.     

SKP2 regulates ZEB1 expression and stimulates eutopic endometrial stromal cell invasion and proliferation of adenomyosis

Though endometriosis is benign, however, it shares certain characteristics with cancers, such as the ability to invade and metastasize. Previous studies have demonstrated that S-phase kinase associated protein2 (SKP2) promotes invasion, tumorigenesis, and metastasis. However, its correlation with adenomyosis is unclear. Herein, we aimed to look into SKP2 expression patterns and regulatory effects on endometrial stromal cell (ESC) proliferation and invasion, and its internal mechanism in adenomyosis. Western blot, qRT-PCR, and immunochemistry were carried out for detecting SKP2 and ZEB1 expression in ESC of adenomyosis and adenomyosis endometrial tissue. The primary ESCs were identified using immunofluorescence. SKP2 knockdown was accomplished in vitro by transfecting a particular lentivirus vector. The colony formation and CCK-8 assays were carried out for assessing cell proliferation, while cell invasion potential was assessed using the transwell assay. Both SKP2 and ZEB1 were found to be significantly upregulated in adenomyosis endometrial tissue. Knockdown of SKP2 inhibited adenomyotic ESC invasion and proliferation. Further experiments showed that knocking out SKP2 reduced ZEB1 expression in adenomyotic ESCs. Our results showed that SKP2 could regulate ZEB1 expression, and increased SKP2 may play a role in the pathogenesis of adenomyosis and stimulating ESC proliferation and invasion.     

Increase of DNA synthesis in uterine adenomyosis in mice with ectopic pituitary isograft.

Ectopic pituitary isografts (EPI) have been found to induce a high incidence of uterine adenomyosis in SHN mice. All the SHN mice given EPI in the right uterus at 40 days of age developed uterine adenomyosis, and more than 80% of mice showed the genesis of subserosal nodules, an advanced state of adenomyosis, 65 days after EPI. Activities of both thymidylate synthetase and thymidine kinase, i.e. DNA-synthesizing enzymes in de novo and salvage pathways of pyrimidine metabolism, respectively, were significantly increased in EPI-induced uterine adenomyosis to approximately 2-fold those in normal control uteri. Bromodeoxyuridine-immunoreactive cells were regarded as the cells in S phase, and the number in the endometrial epithelium and stroma in EPI-induced uterine adenomyosis was more than 1.5-fold that in normal control uteri. EPI may affect the genesis of uterine adenomyosis generally, but not locally, because there were no differences between the right uterus with EPI and the left without EPI in the incidence of adenomyosis, histology or DNA-synthesizing enzyme activities.     

Oestrogen‐induced angiogenesis promotes adenomyosis by activating the Slug‐VEGF axis in endometrial epithelial cells

Adenomyosis is an oestrogen‐dependent disease characterized by the invasion of endometrial epithelial cells into the myometrium of uterus, and angiogenesis is thought to be required for the implantation of endometrial glandular tissues during the adenomyotic pathogenesis. In this study, we demonstrate that compared with eutopic endometria, adenomyotic lesions exhibited increased vascularity as detected by sonography. Microscopically, the lesions also exhibited an oestrogen‐associated elevation of microvascular density and VEGF expression in endometrial epithelial cells. We previously reported that oestrogen‐induced Slug expression was critical for endometrial epithelial–mesenchymal transition and development of adenomyosis. Our present studies demonstrated that estradiol (E2) elicited a Slug‐VEGF axis in endometrial epithelial cells, and also induced pro‐angiogenic activity in vascular endothelial cells. The antagonizing agents against E2 or VEGF suppressed endothelial cells migration and tubal formation. Animal experiments furthermore confirmed that blockage of E2 or VEGF was efficient to attenuate the implantation of adenomyotic lesions. These results highlight the importance of oestrogen‐induced angiogenesis in adenomyosis development and provide a potential strategy for treating adenomyosis through intercepting the E2‐Slug‐VEGF pathway.