子宫内膜异位症患者在位内膜及异位内膜中Claudin-1、Claudin-3的表达及意义

目的:探讨紧密连接蛋白claudin-1、claudin-3在子宫内膜异位症(内异症)发生发展中的作用及意义.方法:免疫组织化学方法检测正常子宫内膜、内异症患者的在位内膜和异位内膜组织中claudin-1、claudin-3蛋白质的表迭.结果:chudin-1、claudin-3蛋白主要定位于子宫内膜细胞的细胞膜/质,内异症患者的异位内膜中claudin-1、claudin-3蛋白的表达显著低于在位内膜及正常子宫内膜(P＜0.01).结论:claudin-1、claudin-3蛋白在内异症患者异住内膜中的表达明显下调,二者可能与内异症的发生发展相关.     

趋化因子CCR7受体在子宫内膜组织上的表达水平及意义

目的通过观察趋化因子CCR7受体在子宫内膜异位症组织上的表达,探讨趋化因子CCR7受体在子宫内膜异位症发病中的作用。方法采用SABC免疫组化染色法观察CCR7在正常对照的子宫内膜和AM的异位子宫内膜及OEM的异位子宫内膜、在位子宫内膜腺上皮组织中表达水平。结果 CCR7主要在腺上皮细胞的胞浆和胞膜表达,CCR7受体的表达水平在AM组及OEM组的异位、在位子宫内膜显著高于正常子宫内膜(P0.05)。CCR7受体的表达水平在AM组及OEM组的异位内膜间差异无统计学意义(P0.05)。结论     

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。     

P物质在子宫内膜异位症中的表达及其意义

目的:探讨P物质(substance P,SP)在子宫内膜异位症(endometriosis,EM)中的表达及其意义。方法:采用免疫组织化学法检测2012年10月至2013年4月在哈尔滨医科大学附属第一医院经腹腔镜及病理证实的EM患者异位子宫内膜组织20例,与其配对的在位子宫内膜组织10例以及因非EM(子宫肌瘤)行腹腔镜下子宫全切术或肌瘤核除患者的正常子宫内膜组织20例中SP的表达情况,并分析和比较术中盆腔粘连发生情况。结果:SP在EM患者的异位子宫内膜、在位子宫内膜及非EM患者的正常子宫内膜的阳性表达率分别为75%、80%、20%,异位子宫内膜和在位子宫内膜比较无显著性差别(P=1.0),但均高于非EM患者的正常子宫内膜,差异均有统计学意义(P=0.002,P=0.004);EM组盆腔粘连的阳性率高于非EM组,EM患者异位子宫内膜中SP阳性组盆腔粘连阳性率高于SP阴性组,差异均有统计学意义(P=0.001,P=0.032),非EM患者正常子宫内膜中SP阳性组盆腔粘连阳性率与SP阴性组相比,差异不具有统计学意义(P=0.061)。结论:SP在EM的异位子宫内膜和在位子宫内膜中的表达上调,并与EM合并盆腔粘连有关,其具体机制尚有待进一步的研究。     

原因不明子宫内膜薄雌激素受体α基因多态性及其表达

旨在探讨原因不明子宫内膜薄雌激素受体α(estrogen receptor alpha,ERα)基因多态性及其与表达的关系。选择120名原因不明子宫内膜薄患者为试验组,120名子宫内膜正常人群作为对照组。应用分子生物学的方法分析ERα基因PvuⅡ,XbaⅠ限制性片段长度多态性。通过逆转录-多聚酶链反应(RT-PCR)和Western印迹法分析ERα表达。结果显示,P基因型频率试验组为47.1%,对照组为30.0%,OR值:2.076。试验组X基因型频率为20.8%,对照组为30.4%,OR值:0.602。Pvu II和Xba I限制性片段长度多态性在两组中均呈多态性分布。试验组ERα的mRNA和蛋白质表达均比对照组降低(P0.05)。由此得出,ERα基因多态性与原因不明子宫内膜薄有关,P等位基因可能是其危险因素,X等位基因可能是其保护因素。ERα在子宫内膜中原因不明子宫内膜薄中的表达低于子宫内膜厚度正常子宫内膜。     

非动情期SD大鼠子宫内膜异位症模型的建立

目的　研究SD大鼠非动情期子宫内膜异位症建立方法和异位病灶的组织学形态及体液免疫反应。方法　采用外科诱导法对 40只非动情期和 40只动情期SD雌性大鼠行子宫组织自体移植手术。采集造模前后的血液进行体液免疫IgG、IgA、IgM、C3 、C4 的观察 ;手术后 7周取健侧子宫中段的在位内膜和移植的异位内膜行组织学研究。结果　SD大鼠非动情期进行子宫组织自体异位种植术成模率 95% ,与大鼠动情期进行子宫组织自体异位种植术成模率 94 8%比较 ,二者差异无显著性 ;异位内膜上皮细胞功能活跃 ,有类似正位子宫内膜的周期变化 ,其生理特征与正位子宫内膜基本相同。且造模后大鼠体液免疫反应敏感IgG、C3 增高 (P <0 0 1、3 P <0 0 5) ;IgM降低 (P <0 0 1)、IgA、C4 差异无显著性。结论　SD大鼠非动情期和动情期进行子宫组织自体异位种植术均可建立良好的子宫内膜异位症模型     

Musashi-1和β-catenin在子宫内膜中的表达及其在内膜异位病灶形成中的作用

目的:检测Musashi-1和β-连环蛋白(β-catenin)在子宫内膜异位症(EMs)患者的在位内膜和异位内膜中的表达,并初步探讨作用机制。方法:2016年9月至2018年9月,收集EMs患者的在位内膜(在位内膜组,28例)、异位内膜(异位内膜组,24例)和非EMs患者的正常内膜(正常内膜组,30例),采用实时荧光定量PCR(Real-time PCR)试验检测Musashi-1和β-catenin在各组内膜组织中的表达情况,并分析Musashi-1和β-catenin在各组内膜组织中的表达相关关系。结果:在位内膜组和异位内膜组中,Musashi-1和β-catenin的相对表达量均明显高于正常内膜组(P0.05),而在位内膜组与异位内膜组之间的差异无统计学意义(P0.05)。在位内膜组和正常内膜组中,增生期的Musashi-1和β-catenin相对表达量显著高于分泌期(P0.05),而异位内膜组中,在增生期和分泌期Musashi-1和β-catenin比较差异无统计学意义(P0.05)。在位内膜组和异位内膜组中,Musashi-1和β-catenin表达之间均呈正相关性(P0.05),而正常内膜组中,两者表达之间无明显相关性(P0.05)。结论:在EMs的发病过程中,干细胞标志物Musashi-1可能通过激活Wnt/β-catenin信号通路参与并促进子宫内膜异位病灶的形成。     

基质金属蛋白酶-2 在子宫内膜异位症中的表达及应用价值

目的:探讨粘附分子CD44拼构变异体6(CD44v6)和基质金属蛋白酶-2(MMP-2)在子宫内膜异位症(EMs)组织中的表达及相关性。方法:选取20例异位内膜组织标本、20例在位内膜组织标本及20例正常子宫内膜标本,用病理常规免疫组织化学方法检测MMP-2和CD44v6的表达,并分析其相关性。结果:CD44v6在异位内膜组的表达明显高于在位内膜组和对照组,且对照组明显高于在位内膜组,差异具有统计学意义(P0.05);CD44v6在在位内膜组和对照组中分泌期的表达明显高于同组增生期,差异具有统计学意义(P0.05)。MMP-2在异位内膜组和在位内膜组的表达明显高于对照组,差异具有统计学意义(P0.01);MMP-2在各组增生期和分泌期表达不规律。异位内膜组中,CD44v6和MMP-2在Ⅲ-Ⅳ期的表达明显高于Ⅰ-Ⅱ期,差异具有统计学意义(P0.01)。Spearman相关性分析结果显示:EMs组织中MMP-2和CD44v6之间呈现正相关性(r=0.724,P0.05);EMs不同分期组织中MMP-2和CD44v6之间亦呈现正相关性(r=0.623,P0.05)。结论:MMP-2和CD44v6在EMs异位内膜中高表达,且有正协同作用,二者可能与EMs的发生发展有关。     

上皮间质转化标记物及Snail在子宫内膜异位症中的表达

目的:研究上皮间质转化标志物(E-cadherin、β-catenin、vimentin)和Snail在子宫内膜异位症(endometriosis,EMs)中的表达。方法:选取40例EMs患者(实验组)异位内膜及在位内膜,同时获取20例非EMs患者(对照组)的正常子宫内膜,采用免疫组化法研究Snail、EMT上皮标志物(E-cadherin、β-catenin)、间质标志物(vimentin)在各内膜组织中的表达,并比较其表达水平。结果:EMs患者异位内膜和在位内膜的EMT上皮标志物E-cadherin、β-catenin表达均显著低于正常内膜的表达(P0.05);EMs患者异位内膜和在位内膜的EMT间质标志物vimentin表达均显著高于正常内膜的表达(P0.05);EMs患者异位内膜和在位内膜中Snail表达显著高于正常内膜的表达(P0.05)。结论:在子宫内膜异位症(EMs)中,Snail、vimentin表达上调,E-cadherin、β-catenin表达下调可能与子宫内膜异位症(EMs)的发生、发展及浸润转移有关。     

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.     

YKL-40 和NF-κ B 在子宫内膜异位症中的表达及相关性研究

目的:通过检测人类软骨蛋白39(YKL-40)和核转录因子KappaB(NF-κB)在子宫内膜异位症组织中的表达,探讨二者与子宫内膜异位症的关系及二者的相关性。方法:用免疫组化二步法检测40例子宫内膜异位症(EMS)患者的异位内膜和在位内膜及40例子宫肌瘤患者子宫内膜(对照组)YKL-40和NF-κB的表达,并对检测结果进行统计学分析。结果:YKL-40、NF-κB在子宫内膜异位症患者的异位和在位内膜及对照组内膜中的表达率分别为67.5%、62.5%、35%及88.5%、57.5%、32.5%,差异有统计学意义(P<0.01);在不同月经周期YKL-40表达无统计学意义;在位内膜和正常内膜NF-κB的表达分泌期高于增殖期,差异有统计学意义(P<0.05);YKL-40和NF-κB在三种内膜中的表达具有正相关性,相关系数分别是0.305,0.267和0.457(P<0.01)。结论:YKL-40和NF-κB在EMS发生发展中起重要的作用。     

Dominant expression of progesterone receptor form B mRNA in ovarian endometriosis

This study was designed to examine the biological implications of progesterone receptor form A (PR-A) and B (PR-B) mRNA expressions in human ovarian endometriosis (ectopic endometrium). A high ratio of PR-B to PR-AB (PR-A+PR-B) mRNA expression was found in 8 of 14 cases of endometriosis, compared with the ratio in eutopic endometrium. The mean ratio in ectopic endometria was significantly (p < 0.01) higher than in eutopic endometria. The ratio in eutopic and ectopic endometria showed no significant change during the menstrual cycle. The mean ratio in ectopic endometria in the proliferative and secretory phases of the endometrium was significantly (p < 0.01) higher than in eutopic endometria. In conclusion, PR-B mRNA was relatively highly expressed in some endometriomas, which might lead to aberrations in the control of progestational effects involving responsiveness to sex steroidal growth regulation.     

Characterization of periostin expression in human endometrium and endometriotic lesions

Objective(s): To investigate the expression of periostin in the eutopic and ectopic endometrium of women diagnosed as endometriosis and evaluate the role of periostin in the pathogenesis of endometriosis. Study design: In this study, the expression of periostin was evaluated in the endometrial specimens from 35 women diagnosed as endometriosis and from 30 healthy women. To assess the presence and localization of periostin throughout the menstrual cycle in both eutopic and ectopic endometrium of women with endometriosis, microscopic evaluation was conducted. It was also subsequently compared with normal endometrium. Results: In the eutopic and ectopic endometrium of women with endometriosis, immunoreactivities of periostin increased compared with those of normal endometrium. We also observed a cyclic variation in the eutopic stromal periostin immunoreactivity throughout their menstrual cycle because higher H score values were observed in the proliferative phase than those in the secretory phase. Conclusion(s): These findings indicated that periostin may be involved in the pathophysiology of endometriosis.     

Differential Expression of CRH,UCN, CRHR1 and CRHR2 in Eutopic and Ectopic Endometrium of Women with Endometriosis

Endometriosis is considered as a benign aseptic inflammatory disease, characterised by the presence of ectopic endometrium-like tissue. Its symptoms (mostly pain and infertility) are reported as constant stressors. Corticotropin releasing hormone (CRH) and urocortin (UCN) are neuropeptides, strongly related to stress and inflammation. The effects of CRH and UCN are mediated through CRHR1 and CRHR2 receptors which are implicated in several reproductive functions acting as inflammatory components. However, the involvement of these molecules to endometriosis remains unknown. The aim of this study was to examine the expression of CRHR1 and CRHR2 in endometriotic sites and to compare the expression of CRHR1 and CRHR2 in eutopic endometrium of endometriotic women to that of healthy women. We further compared the expression of CRH, UCN, CRHR1 and CRHR2 in ectopic endometrium to that in eutopic endometrium of women with endometriosis. Endometrial biopsy specimens were taken from healthy women (10 patients) and endometrial and endometriotic biopsy specimens were taken from women with endometriosis (16 patients). Τhe expression of CRH, UCN, CRHR1, and CRHR2 was tested via RT-PCR, immunohistochemistry and Western blotting. This study shows for the first time that CRH and UCN receptor subtypes CRHR1β and CRHR2α are expressed in endometriotic sites and that they are more strongly expressed (p<0.01) in eutopic endometrium of women with endometriosis compared to healthy women endometrium at the mRNA and protein level. CRH, UCN, CRHR1 and CRHR2 mRNA were also more highly expressed in ectopic rather than eutopic endometrium (CRH, UCN, CRHR2α: p<0.01, CRHR1β: p<0.05) and protein (CRH and UCN: p<0.05, CRHR1 and CRHR2: p<0.01) in women with endometriosis. These data indicate that CRH and UCN might play an immunoregulatory role in endometriotic sites by affecting reproductive functions such as decidualization and implantation of women with endometriosis.     

Galectin-1 Overexpression in Endometriosis and Its Regulation by Neuropeptides (CRH,UCN) Indicating Its Important Role in Reproduction and Inflammation

Endometriosis is an inflammatory disease of women of reproductive age featured by the presence of ectopic endometrium and is strongly related to infertility. Galectins, carbonhydrate-binding proteins, have been found to have pro- or anti-inflammatory roles in the reproductive tract and in pathological conditions concerning infertility. Galectin-1, which is expressed at endometrium and decidua, plays a major role in implantation and trophoblast invasion. Also, the neuropeptides, corticotropin releasing hormone (CRH) and urocortin (UCN) and their receptors are expressed in eutopic and ectopic endometrium showing a differential expression pattern in endometriotic women compared to healthy ones. The aim of this study was to examine the galectin-1 expression in endometriotic lesions and compare its expression in eutopic endometrium of endometriotic and healthy women. Furthermore, we examined the effect of CRH and UCN in galectin-1 expression in Ishikawa cell line and macrophages and investigated the implication of CRHR1 in these responses. Eutopic and ectopic endometrium specimens, Ishikawa cell line and mice macrophages were used. Immunohistochemistry and western blot analysis were performed in order to identify galectin-1 expression in ectopic and eutopic endometrium of women with and without endometriosis and the regulatory effect of CRH and UCN on galectin-1 expression. This study presents for the first time that galectin-1 is overexpressed in endometriotic lesions compared to eutopic endometrium of endometriotic women and is more abundantly expressed in eutopic endometrium of disease women compared to healthy ones. Furthermore, it is shown that CRH and UCN upregulate galectin-1 expression in Ishikawa cell line and macrophages and this effect is mediated through CRHR1. These results suggest that galectin-1 might play an important role in endometriosis pathology and infertility profile of women suffering from endometriosis by being at the same time regulated by CRH and UCN interfering in the immune disequilibrium which characterizes this pathological condition.     

YKL-40和NF—KB在子宫内膜异位症中的表达及相关性研究

目的：通过检测人类软骨蛋白39（YKL-40）和核转录因子KappaB（NF—KB）在子宫内膜异位症组织中的表达,探讨二者与子宫内膜异位症的关系及二者的相关性。方法：用免疫组化二步法检测40例子宫内膜异位症（EMS）患者的异位内膜和在位内膜及40例子宫肌瘤患者子宫内膜（对照组）YKL-40和NF—KB的表达,并对检测结果进行统计学分析。结果：YKL-40、NF—KB在子宫内膜异位症患者的异位和在位内膜及对照组内膜中的表达率分别为67．5％、62．5％、35％及88．5％、57．5％、32．5％,差异有统计学意义（P〈0．01）;在不同月经周期YKL-40表达无统计学意义;在位内膜和正常内膜NF-KB的表达分泌期高于增殖期,差异有统计学意义（P〈0．05）;YKL-40和NF-KB在三种内膜中的表达具有正相关性,相关系数分别是0．305,0．267和0．457（P〈0．01）。结论：YKL-40和NF—KB在EMS发生发展中起重要的作用。     

Aberrant methylation of the IL‐12B promotor region contributes to the risk of developing ovarian endometriosis

Studies have shown that aberrant expression of IL‐12p40, which is encoded by the interleukin‐12B (IL‐12B) gene, may be involved in the development of endometriosis. In this study, we investigated the role of aberrant methylation of the IL‐12B promoter region and its associated expression in the development of ovarian endometriosis. By using pyrosequencing, we analyzed the methylation level of the IL‐12B promoter region in eutopic and ectopic endometrium of patients with ovarian endometriosis and normal endometrium of control women. The expression of IL‐12B mRNA was detected by quantitative real‐time PCR. The results showed that the methylation level of the IL‐12B promoter region in ectopic and eutopic endometrium of patients with ovarian endometriosis was significantly lower than that in endometrium of women without endometriosis ( p < 0.001 and p = 0.041, respectively). In contrast, mRNA levels were significantly increased in ectopic and eutopic endometrium of patients with ovarian endometriosis compared to those in endometrium of women without endometriosis ( p < 0.001 and p = 0.042, respectively). Correlation analysis showed that the methylation level of the IL‐12B promoter region was negatively correlated with mRNA levels of IL‐12B ( p < 0.001). Our data suggested that aberrant methylation of the IL‐12B promoter region may be responsible for aberrant IL‐12B mRNA expression in endometrium tissue of women, which may be associated with the development of ovarian endometriosis in northern Chinese women.     

miRNA的重要调节者Dicer和Drosha与子宫内膜异位症的相关性研究

利用荧光定量PCR和Western blot检测证实,在异位的子宫内膜组织中Dicer和Drosha的表达低于在位子宫内膜组织,随后体外培养在位子宫内膜组织,采用siRNA干扰Dicer和Drosha,发现与干扰对照组相比,子宫内膜细胞的增殖加快而凋亡减少;同时ELISA检测显示转化生长因子-β1(transforming growth factor-beta 1, TGF-β1)的表达上调;Western blot检测显示凋亡抑制蛋白Bcl2表达增加而促凋亡蛋白Bax的表达减少．结果表明,miRNA的重要调节者Dicer和Drosha可以影响TGF-β1及Bcl2/Bax的表达进而影响细胞的增殖和凋亡,从而参与了子宫内膜异位症的形成．