【作者】 王茹娜；

【导师】 孟跃进；

【作者基本信息】 郑州大学 ， 妇产科学， 2013， 硕士

【摘要】 背景和目的宫腔粘连(intrauterine adheious IUA)是由于宫腔受到损伤或感染导致子宫内膜基底层受损,无法再生修复,宫腔内瘢痕形成而致宫腔形态失常,约90%由过度刮宫引起。近年来,随着宫腔操作机率的增加和宫腔镜技术的发展,IUA患病率呈逐年上升趋势,其发生严重影响现代妇女的生活质量及生育要求。目前,其发病机制尚不完全明确,可能与体内部分促进创面修复或抑制组织纤维化的细胞因子异常表达密切相关。为此,探讨TGF-β1、Smad3、IFN-γ在宫腔粘连中的表达,对于了解IUA的发病机制、为预防IUA的形成及术后复发、应用抗TGF-β抗体和TGF-β1抑制剂抗纤维化治疗提供重要的理论依据。转化生长因子-β1(Transfer Growth Factor, TGF-β1)是目前公认的最强的促纤维化因子,是致组织纤维瘢痕形成通路中的关键介导因子。TGF-β1作为多功能细胞因子,通过与一些相关的炎性细胞因子相互作用,成纤维细胞大量增殖,细胞外基质过度沉积致组织纤维化；此外,TGF-β1通过对血管内皮细胞的趋化迁移及血管内皮生长因子(VEGF)的诱导作用,促进瘢痕组织血管的形成。Smads蛋白家族主要包括Smad1～10, Smad3蛋白是将TGF-β活化并与受体结合产生的信号由胞质转导至胞核内的重要信号转导分子,通过TGF-β1/Smad3通路作用致胶原合成增加和细胞外基质沉积,促使组织纤维瘢痕形成。干扰素-γ(interferon-gamma,IFN-γ)是目前比较明确的抑制组织纤维化的调节因子,具有影响细胞生长和分化以及免疫调节等多种生物学功能。IFN-y主要通过JAK/STAT途径促进Smad7表达的显著增加,造成TGF-β1/Smad3通路中信号的传递过程受阻,抑制TGF-β1和Smad3蛋白的表达,减少细胞外基质的沉积。材料与方法1.实验分组研究组：选择2012年5月至2012年10月在我院行宫腔镜下宫腔粘连分离术的中重度宫腔粘连患者30例。依据取材部位的不同分为A组(粘连组织)和B组(靠近粘连的内膜组织)。正常对照组：选择同期因不孕行宫腔镜检查且子宫内膜正常的非IUA患者30例,取其内膜,病理结果为增生期子宫内膜。2.实验方法采用免疫组织化学SP法分别检测TGF-β1、Smad3、IFN-γ在粘连组织、粘连旁内膜组织及非IUA患者正常内膜组织中的表达。以α=0.05为检验水准。3.统计方法全部实验数据输入SPSS17.0软件包处理。TGF-β1、Smad3、IFN-γ分别在粘连组织、粘连旁内膜组织及非IUA患者正常内膜组织中的表达采用单因素方差分析,组间两两采用LSD-t检验,各个因子之间的关联分析采用Pearson相关分析。结果1.TGF-β1的表达(1) TGF-β1在粘连组织中的表达高于粘连旁内膜组织,差异有统计学意义(P<0.05)(2) TGF-β1在粘连组织中的表达高于非IUA正常内膜组织,差异有统计学意义(P<0.05)(3) TGF-β1在粘连旁内膜组织中的表达高于非IUA正常内膜组织,差异有统计学意义(P<0.05)2.Smad3的表达(1) Smad3在粘连组织中的表达高于粘连旁内膜组织,差异有统计学意义(P<0.05)(2) Smad3在粘连组织中的表达高于非IUA正常内膜组织,差异有统计学意义(P<0.05)(3)Smad3在粘连旁内膜组织中的表达高于非IUA正常内膜组织,差异有统计学意义(P<0.05)3.IFN-γ的表达(1) IFN-γ在粘连组织中的表达低于粘连旁内膜组织,差异有统计学意义(P<0.05)(2) IFN-γ在粘连组织中的表达低于非IUA正常内膜组织,差异有统计学意义(P<0.05)(3) IFN-γ在粘连旁内膜组织中的表达低于非IUA正常内膜组织,差异有统计学意义(P<0.05)4.TGF-β1与Smad3蛋白在宫腔粘连组织及粘连旁内膜组织中的表达成正相关。(P<0.05,r1=0.96,r2=0.39)5.TGF-β1与IFN-γ蛋白在宫腔粘连组织及粘连旁内膜组织中的表达成负相关。(P<0.05r1=-0.98,r2=-0.97)结论1.TGF-β1与Smad3在粘连组织及粘连旁内膜组织均呈高表达。且二者呈正相关,共同促进宫腔粘连的形成与发展。2.IFN-γ在粘连组织及粘连旁内膜组织的表达均低于非IUA正常内膜中的表达,与TGF-β1呈负相关,其低表达与宫腔粘连形成相关。3.以TGF-β1、Smad3和IFN-γ在TGF-β1/Smad3通路中的关系为出发点,为IUA的靶向治疗提供理论依据。更多还原

【Abstract】 Background and PurposeIntrauterine adhesions is caused by injuries or infections that induce intrauterine endometrium base layer damaged and irregenerative and intrauterine scar formation.These lead to the form of intrauterine abnormal which about90%is due to excessive curettage.In recent years,with the increase that the chance of uterine cavity operation and the development of modern hysteroscopy technology,its prevalence rate is increasing year by year,it seriously affected the modern women’s quality of life and fertility requirements. So far the exact pathogenesis of Intrauterine adhesions is uncertain.It may be related to the abnormal expression of some cytokines promoting wound repair or inhibition of tissue fibrosis.Therefor,it is an important theoretical basis that to discuss the expression of TGF-β1、Smad3and IFN-γ in the fibrosis tissue and endomertrium near the fibrosis of patients with intrauterine adhesions,for the pathogenesis of Intrauterine adhesions,prevent the formation of IUA and postoperative recurrence and use of TGF-beta antibodies and TGF-beta1inhibitor anti fibrosis treatmentNow,TGF-β1is recognized as the strongest fibrosis factors,and it is the key mediating factor of fibrous scar formation pathways. TGF-β1as a kind of multifunctional cytokine induce tissue fibrosis, through interaction associated with some inflammatory cytokines, fibroblast proliferation in great quantities, excessive extracellular matrix deposition.Moreover, TGF-beta1through to chemotaxis migration of the vascular endothelial cell and entrainment of small tube skin growth factor (VEGF), promote blood vessels of the formation of scar tissue.Smads proteins family including Smad1～10.Smad3protein is an important signal transduction molecule which transduces the signal originated from combination of TGF-beta activated and receptors from cytoplasmic to nuclei.Through the TGF-beta1/Smad3pathways to increase collagen synthesis and deposition of extracellular matrix, tissue fibrous scar formation.It is the more cleat that IFN-gamma is the adjustment factor of restrain tissue fibrosis,It has a variety of biological functions,such as influence cell growth and differentiation,and immune regulation.Causes of TGF-beta1/Smad3pathway of signal in transmission process is blocked, inhibited TGF-beta1and Smad3protein expression,and reduce the deposition of extracellular matrix.Material and Methods1.GroupingThe experimental group:Take the fibrosis tissue and endomertrium near the fibrosis tissue in30patients dignosed as intrauterine adheious in department of gynaecology and obstetrics of the second affiliated hospital of Zhengzhou University from May2012to October2012.The control group:Take normal endometrium in30infertility patients who were accepted hysteroscopic examination in depart tment of gynaecology and obstetrics of the second affiliated hospital of Zhengzhou University from May2012to October2012.Take normal intima proved by pathology.2.MethodsUse Immunohistohistochemistry SP method to detect the expression of TGF-beta1, Smad3and IFN-γ in three tissue.3.Statistical analysisUsing SPSS package for windows17.0statistical analysis to test all data.The expression of TGF-beta1, Smad3and IFN-y in three tissue is compared by Oneway ANOVA;Different binary comparison between the groups uses the LSD-t test.The correlation between groups is tested by Pearson correlation analysis.The level for significance tests is0.05.Results1. Expression of TGF-beta1(1)The expression of TGF-betal in the fibrosis tissue is higher than that in endometrium near the fibrosis tissue.The difference is significant statistically (P<0.05).(2)The expression of TGF-beta1in the fibrosis tissue is higher than that in normal endometrium tissue.The difference is significant statistically(P<0.05).(3)The expression of TGF-beta1in endometrium near the fibrosis tissue is higher than that in normal endometrium tissue.The difference is significant statistically(P<0.05).2.Expression of Smad3(1)The expression of Smad3in the fibrosis tissue is higher than that in endometrium near the fibrosis tissue.The difference is significant statistically (P<0.05).(2)The expression of Smad3in the fibrosis tissue is higher than that in normal endometrium tissue.The difference is significant statistically(P<0.05).(3)The expression of Smad3in endometrium near the fibrosis tissue is higher than that in normal endometrium tissue.The difference is significant statistically (P<0.05).3.Expression of IFN-γ(1)The expression of IFN-γ in the fibrosis tissue is lower than that in endometrium near the fibrosis tissue.The difference is significant statistically (P<0.05).(2)The expression of IFN-γ in the fibrosis tissue is lower than that in normal endometrium tissue.The difference is significant statistically(P<0.05).(3)The expression of IFN-γ in endometrium near the fibrosis tissue is lower than that in normal endometrium tissue.The difference is significant statistically(P<0.05).4.Relationship between TGF-beta1and Smad3 There is a positive correlation between the expression of TGF-betal and the expression of Smad3in the fibrosis tissue and in endometrium near the fibrosis tissue.5.Relationship between TGF-betal and IFN-γThere is a negative correlation between the expression of TGF-betal and the expression of IFN-γ in the fibrosis tissue and in endometrium near the fibrosis tissue.Conclusions1.TGF-betal and Smad3are higher expressions in the fibrosis tissue and in endometrium near the fibrosis tissue,which related to the formation of Intrauterine adhesions.2.IFN-γ is lower expression in the fibrosis tissue and in endometrium near the fibrosis tissue.There is a negative correlation between the expression of TGF-betal and the expression of IFN-γ, which related to the formation of Intrauterine adhesions.3.Through the relationship of TGF-beta1, Smad3and IFN-γ in TGF-beta1/Smad3pathways, the anti fibrosis treatment is likely to be used in the treatment of uterine cavity adhesion.更多还原