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子宫内膜异位症患者在位内膜中LN及uPAR的检测及其意义

The Detection of LN and uPAR in the Eutopic Endometrium of Patients with Endometriosis and Its Significance

作者: 专业:妇产科学 导师:朱新群 年度:2010 学位:硕士  院校: 郑州大学

Keywords

Endometriosis, Adenomyosis, uterus eutopic endometrium, Laminin, urokinase type Plasminogen Activator Receptor, Enzyme Linked Immunosorbent Assay

        背景与目的子宫内膜异位症(Endometriosis, EMs)是常见多发的良性病变,却具有类似恶性肿瘤的生物学特征。目前EMs术前确诊率低,作为诊断金标准的腹腔镜检查具有创性、步骤繁、价格高等不足,EMs治疗方法不统一且弊端多。为EMs临床提供合适的筛查及干预项目十分必要。EMs异位病灶形成的第一步是内膜从原组织脱离,层粘连蛋白(Laminin, LN)属于黏附分子整合素蛋白家族,是细胞外基质(Extracelluar matrix, ECM)的主要成分及细胞黏附于基质的重要介质;EMs异位病灶形成的关键环节是ECM的降解,尿激酶受体(urokinase type plasminogen activator receptor, uPAR)是胞外纤溶酶系统的重要一员,其可介导纤溶酶活化、ECM组分降解及组织细胞侵袭生长等。有关不同个体(EMs患者与非EMs者)的子宫内膜,是否存在LN与uPAR水平的差异及这种差异可能产生的临床价值,目前国内外文献报道尚少。本研究针对该问题作一些初步探讨。材料与方法1实验分组留取的子宫内膜组织标本来源于2008年2月~2009年2月郑州大学第二附属医院妇产科住院行手术治疗的患者84例,盆腔EMs组28例(其中Ⅰ~Ⅱ期EMs 12例,Ⅲ~Ⅳ期EMs 16例)、子宫腺肌症组20例及非EMs组36例。2实验方法采用酶联免疫吸附法检测内膜组织匀浆样品中LN/uPAR的OD值,使用Bradford法测定样品的蛋白浓度,从而得到样品中LN/uPAR的蛋白含量。3统计分析方法应用SPSS13.0软件进行统计分析,所有数据以均数±标准差表示。组间比较采用单因素方差分析;盆腔EMs组内比较采用成组资料t检验;相关性检验采用Pearson等级相关进行分析。统计方法均以α=0.05作为检验水准。结果1三组资料子宫内膜匀浆液中LN水平比较非EMs者的子宫内膜、子宫腺肌症及盆腔EMs患者的在位内膜匀浆液中LN水平分别为61.02±7.45μg/mg蛋白、53.03±7.22μg/mg蛋白和40.95±9.10μg/mg蛋白,三组总体比较,差异有统计学意义(F=24.895,P=0.001<0.05);两两比较,差异均有统计学意义(P<0.05)。Ⅰ~Ⅱ期EMs与Ⅲ~Ⅳ期EMs患者的LN水平分别为47.82±6.93μg/mg蛋、白和35.61±6.68μg/mg蛋白,两组比较,差异具有统计学意义(T=5.034,P)=0.001<0.05)。2三组资料子宫内膜匀浆液中uPAR水平比较非EMs者的子宫内膜、子宫腺肌症及盆腔EMs患者的在位内膜匀浆液中uPAR水平分别为3.77±0.71ng/mg蛋白、4.30±0.47ng/mg蛋白和5.28±0.62ng/mg蛋白,三组总体比较,差异有统计学意义(F=30.283,P=0.001<0.05);两两比较,差异均有统计学意义(P<0.05)。Ⅰ~Ⅱ期EMs与Ⅲ-Ⅳ期EMs患者的uPAR水平分别为5.45±0.62ng/mg蛋白和5.06±0.55ng/mg蛋白,两组比较,差异有统计学意义(T=-2.312,P=0.025<0.05)3子宫腺肌症中LN与uPAR水平相关性分析子宫腺肌症患者在位内膜的匀浆液中LN与uPAR水平之间无相关性(rp=0.154,P=0.518>0.05)。4盆腔EMs中LN与uPAR水平相关性分析盆腔EMs患者在位内膜的匀浆液中LN与uPAR水平之间无相关性(rp=0.079,P=0.669>0.05)。结论1非EMs者的子宫内膜、子宫腺肌症及盆腔EMs患者的在位内膜中LN水平均不同, EMs病情越重,LN水平越低。2非EMs者的子宫内膜、子宫腺肌症及盆腔EMs患者的在位内膜中uPAR水平均不同,且EMs病情越重,uPAR水平越高。3子宫腺肌症及盆腔EMs患者的在位内膜中LN与uPAR水平之间不具有相关性。
    Backgrounds and ObjectivesAlthough Endometriosis is multiple common benign lesions, it has similar biological characteristics of malignant tumors. At present, preoperative diagnosis rate of endometriosis was low, Laparoscopy is the gold standard of the EMs diagnostic, but there is some shortcoming, such as traumatic, cumbersome and expensive procedure, etc, the clinical treatment of EMs is not yet unified, and there are several drawbacks. Providing appropriate clinical screening and intervention programs for EMs is necessary.The first step in the formation of the ectopic lesions of endometriosis is the endometrial cells separated from the original tissue, Laminin (LN) belong to adhesion molecules integrin protein family, the main components of the extracellular matrix (ECM), and important medium of the cell adhesion in the matrixin, after LN and its receptor combined. The key steps in the formation of the ectopic lesions of endometriosis is the degradation of extracellular matrix, urokinase type plasminogen activator receptor (uPAR) is an important member of extracellular plasminogen system, It can mediate the plasminogen activation, ECM degradation and invasive growth of cells in tissue components. Related different levels of LN and uPAR in endometrium of different individuals (endometriosis and non-endometriosis) and the clinical value this difference may produce, currently, there are few reported in the literature. In this study, some of the issues were preliminary studied.Materials and Methods1 Experimental groupsCollected endometrial tissue samples come from 84 cases of surgical treatment hospitalized patients in Second Affiliated Hospital of Zhengzhou University at February 2008 to February 2009,28 cases of pelvic EMs (12 cases of stageⅠ~Ⅱendometriosis,16 cases of stageⅢ~Ⅳendometriosis),20 patients of Adenomyosis group and 36 cases of non-EMs.2 Experimental methodsTo detect the OD values of LN/uPAR in endometrial tissue samples by Enzyme linked immunosorbent assay, followly, determine the protein concentration of samples by Bradford method, to obtain the LN/uPAR protein content in sample.3 Statistical analysis methodsApplicating SPSS 13.0 software conducts statistical analysis. All data are expressed as the form that means±standard deviation. Using single factor analysis of variance test conduct compare among the groups. Using group data t test conduct compare in internal group of Ems. Using Pearson rank correlation conduct analysis of correlation test, and all statistical methods with a= 0.05 as the level of significance.Results1 Compare the LN level in endometrial homogenate of three sets of data.The LN levels in tissue homogenate among the uterus endometrium of non-EMs, eutopic endometrium of patients with adenomyosis and Pelvic EMs were 61.02±7.45μg/mg Protein,53.03±7.22μg/mg Protein and 40.95±9.10μg/mg Protein, respectively. Compare the three groups, overall, the differences were statistically significant (F=24.895, P=0.001<0.05). Comparison was carried out between every two sets of data, the differences were significant all (P<0.05). The LN levels between stageⅠ~ⅡEMs and stageⅢ~ⅣEMs were 47.82±6.93μg/mg Protein and 35.61±6.68μg/mg Protein, respectively. Compare the two groups, the differences were significant (T=5.034, P=0.001<0.05).2 Compare the uPAR level in endometrial homogenate of three sets of data.The uPAR levels in tissue homogenate among the uterus endometrium of non-EMs, eutopic endometrium of patients with adenomyosis and Pelvic EMs were 3.77±0.71 ng/mg Protein,4.30±0.47 ng/mg Protein and 5.28±0.62 ng/mg Protein, respectively. Compare the three groups, overall, the differences were statistically significant (F=30.283, P=0.001<0.05), and and its level of uPAR gradually increased, the uPAR levels were of the highest in III-IV Ems. Comparison was carried out between every two sets of data, the differences were significant (P<0.05). Compare the LN levels between stageⅠ~ⅡEMs and stageⅢ~ⅣEMs were 5.06±0.55 ng/mg Protein and 5.45±0.62 ng/mg Protein, respectively. Compare the two groups, the differences were significant (T=-2.312, P=0.025<0.05).3 Analysis the correlation between LN and uPAR in adenomyosisThe LN and uPAR levels in eutopic endometrial homogenate were not correlated in Patients with adenomyosis (rp=0.154, P=0.518>.05).4 Analysis the correlation between LN and uPAR in Pelvic EMsThe LN and uPAR levels in eutopic endometrial homogenate were not correlated in Patients with Pelvic EMs (rp=0.079, P=0.669>0.05).Conclusions1 The LN levels in tissue homogenate among the Patients with uterus endometrium of non-EMs, eutopic endometrium of patients with adenomyosis and Pelvic EMs were differences, and the more severe EMs condition is, the lowest the LN levels is.2 The uPAR levels in tissue homogenate among the Patients with uterus endometrium of non-EMs, eutopic endometrium of patients with adenomyosis and Pelvic EMs were differences, and the more severe EMs condition is, the highest the uPAR levels is.3 Between the level of LN and uPAR in eutopic endometrium of patients with adenomyosis and Pelvic EMs were not relevant.
        

子宫内膜异位症患者在位内膜中LN及uPAR的检测及其意义

摘要4-7
Abstract7-9
图和附表清单12-13
论文部分 子宫内膜异位症患者在位内膜中LN及uPAR的检测及其意义13-45
    引言13-16
    材料与方法16-24
        1.1 临床资料16-17
        1.2 标本保存17
        1.3 实验仪器和试剂17-18
        1.4 实验方法18-19
        1.5 实验步骤19-23
        1.6 统计分析方法23-24
    结果24-29
        2.1 LN的表达24-26
        2.2 uPAR的表达26-27
        2.3 LN与uPAR水平相关性分析27-29
    讨论29-39
        4.1 不同个体LN水平的差异及其意义29-33
        4.2 不同个体uPAR水平的差异及其意义33-34
        4.3 LN与uPAR的相互关系34-37
        4.4 本研究存在的问题37-38
        4.5 展望38-39
    结论39-40
    附图40-41
    参考文献41-45
综述部分 子宫内膜异位症侵袭性过程相关分子45-58
    参考文献55-58
后记部分58-61
    中英文缩略词对照表58-59
    个人简历59-60
    在学期间发表的学术论文60-61
    致谢61
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