粘附分子VCAM-1和ICAM-1在子宫内膜异位症中的表达及意义

发布时间：2018-05-04 02:39

本文选题：子宫内膜异位症 + VCAM-1　；参考：《浙江大学》2017年硕士论文

【摘要】：研究背景:子宫内膜异位症(简称内异症,endometriosis,EMs)在育龄期妇女中发病率高达10%～15%,近年来发病率有上升趋势。它是指具有生长能力的子宫内膜异位至子宫腔被覆内膜及宫体肌层以外的组织,常常伴有慢性盆腔痛、痛经、不孕等,目前发病机制仍未完全明确。子宫内膜异位症是一种良性疾病,却在侵袭、新生血管生成、增殖、凋亡等诸多方面与恶性肿瘤有极大相似性。现有的研究已经发现了 EMs患者腹腔内环境中的免疫细胞、细胞因子的变化,认为免疫机制异常在EMs的发生、发展过程中起着重要作用。子宫内膜腺上皮及基底膜均有多种细胞粘附分子(cell adhesion molecules,CAMs)的表达,越来越多的研究已证实某些CAMs的异常表达可能参与了 EMs的侵袭、粘附及种植过程。血管细胞粘附分子(vascular cell adhesion molecule 1,VCAM-1)和细胞间粘附分子(intercellular adhesion molecule 1,ICAM-1)是免疫球蛋白超家族家族中的两个重要成员,均为跨膜糖蛋白粘附分子,前者主要分布在激活的内皮细胞、树突状细胞上,只有在体内炎症的病理条件下,才表达于活化的血管内皮上;而后者除了表达在白细胞表面,还可以表达在子宫内膜上皮、间质细胞、血管内皮细胞。因此,我们推测VCAM-1和ICAM-1可能参与了子宫内膜异位症的发生发展过程,对异位内膜的种植、粘附、增殖发挥着重要作用。研究目的:通过检测VCAM-1和ICAM-1在子官内膜异位症(EMs)患者异位内膜、在位内膜、非EMs患者子宫内膜组织和血清中的表达,探讨VCAM-1和ICAM-1在子宫内膜异位发病过程中的作用及临床意义。材料与方法:收集我院盆腔子宫内膜异位症Ⅲ、Ⅳ期(根据r-AFs分期)患者的卵巢异位囊肿壁上的异位子宫内膜作为异位内膜组,其中行宫腔镜检查的子宫内膜作为对应的在位内膜组,选择同期因原发不孕、继发不孕症(输卵管因素,排除内异症)行宫-腹腔镜检查的患者,取其子宫内膜作为对照正常子宫内膜组。此外,选择腹腔镜手术并确诊的盆腔子宫内膜异位症患者术前血清作为血清病例组,选择同期因不孕症(输卵管因素,排除内异症)患者行腹腔镜手术的术前血清作为血清对照组。通过Real-time PCR检测VCAM-1mRNA及ICAM-1 mRNA在不同子宫内膜的表达;通过Western blot方法检测对VCAM-1及ICAM-1蛋白在不同子宫内膜中的表达并进行半定量分析;通过ELISA技术检测内异症患者及对照组血清中sVCAM-1及sICAM-1吸光度(OD值)并进行浓度计算。结果:1.Real-time PCR结果:VCAM-1mRNA在异位子宫内膜组与在位内膜组、对照组之间比较均有显著性差异,具有统计学意义(P0.01,P0.01)。而在位内膜组与对照组比较无统计学意义(P0.05)。ICAM-1mRNA在异位子宫内膜组分别与在位内膜组、对照组比较有显著性差异,具有统计学意义(P0.01,P0.01),而在位子宫内膜组与对照组比较无统计学意义(P0.05)。2.Western blot结果:VCAM-1在异位子宫内膜组、在位内膜组及对照组之间两两比较均有显著性差异,具有统计学意义(P0.01),表达的强弱顺序为:异位内膜组在位内膜组对照组。ICAM-1在异位子宫内膜组分别与在位内膜组、对照组比较有显著性差异,具有统计学意义(P0.01,P0.01),而在位子宫内膜组与对照组比较ICAM-1的相对表达量无统计学意义(P0.05)。3.ELISA结果:根据OD值和标准曲线计算EMs患者血清中的sVCAM-1及sICAM-1平均浓度与对照组比较均明显升高,差异具有统计学意义(P0.01,P0.01)。sICAM-1浓度在Ⅲ、Ⅳ期组比Ⅰ、Ⅱ期组明显升高,差异具有统计学意义(P0.01)。sVCAM-1在早期Ⅰ、Ⅱ期患者的血清浓度高于对照组(P0.01),差异具有统计学意义。采用ROC曲线分析,血清中sICAM-1和sVCAM-1在诊断子宫内膜异位症患者中具有一定的准确性。其中,sICAM-1具有较高的特异性,敏感性偏低。结论:1.VCAM-1和ICAM-1通过在子宫内膜异位症患者中的mRNA水平、组织蛋白水平及血清中的高表达可能参与了子宫内膜异位症的发生、发展,在发病机制中具有重要作用。2.VCAM-1和ICAM-1在子宫内膜异位症患者的异位内膜和在位内膜组织蛋白中的表达无明显相关性,提示两者在子宫内膜异位症的致病机制可能是不同的。3.外周血sICAM-1和sVCAM-1在诊断子宫内膜异位症患者中具有一定的准确性。检测血清sICMA-1的浓度可以了解内异症的进展状况,可能成为诊断内异症的新方法。
[Abstract]:Background: the incidence of endometriosis (endometriosis, EMs) in women of childbearing age is up to 10% to 15%. In recent years, the incidence of endometriosis is rising. It refers to the growth of endometriosis with endometriosis to the endometrium and the tissues outside the intrauterine layer, often with chronic pelvic pain, dysmenorrhea, and infertility. The pathogenesis of endometriosis is still not completely clear at present. Endometriosis is a benign disease, but it has great similarity with malignant tumor in many aspects, such as invasion, angiogenesis, proliferation, apoptosis and so on. Existing studies have found the immune cells in the abdominal environment of EMs patients, the changes of cytokines, and think that the immune mechanism is abnormal in EM The occurrence of s plays an important role in the process of development. There are many cell adhesion molecules (cell adhesion molecules, CAMs) expression in the endometrium epithelium and basement membrane. More and more studies have shown that abnormal expression of certain CAMs may be involved in the invasion, adhesion and planting process of EMs. The adhesion molecules of vascular cells (vascular cell adhesi) On molecule 1, VCAM-1) and intercellular adhesion molecules (intercellular adhesion molecule 1, ICAM-1) are two important members of the immunoglobulin superfamily, all of which are transmembrane glycoprotein adhesion molecules. The former is mainly distributed on activated endothelial cells, dendritic cells, only under the pathological conditions of inflammation in the body to be expressed in activation. In addition to the expression on the white cell surface, the latter can also be expressed in the endometrium epithelium, interstitial cells and vascular endothelial cells. Therefore, we speculate that VCAM-1 and ICAM-1 may be involved in the development and development of endometriosis, and play an important role in the planting, adhesion and proliferation of ectopic endometrium. To detect the expression of VCAM-1 and ICAM-1 in endometriosis (EMs) patients with endometriosis (EMs), eutopic endometrium, the expression of endometrium and serum in non EMs patients, and explore the role and clinical significance of VCAM-1 and ICAM-1 in endometriosis process. Materials and methods: collection of pelvic endometriosis in our hospital, stage III, IV (according to r-AFs staging). The ectopic endometrium on the wall of the ectopic cyst of the ovary of the patient is the ectopic endometrium group, and the endometrium of the hysteroscopy is used as the corresponding eutopic endometrium, and the endometrium of the patients with primary infertility, secondary infertility (fallopian tube factors, excluding the endometriosis) and the endometrium as the normal uterus are chosen at the same time. In addition, the preoperative serum of patients with pelvic endometriosis diagnosed by laparoscopy was selected as a serum case group, and the preoperative serum of patients with infertility (fallopian tube factors excluding endometriosis) underwent laparoscopic surgery as a serum control group. Real-time PCR was used to detect VCAM-1mRNA and ICAM-1 mRNA in different uterus. The expression of the endometrium and the expression of VCAM-1 and ICAM-1 protein in different endometrium by Western blot method and semi quantitative analysis; sVCAM-1 and sICAM-1 absorbance (o value) in serum of patients with endometriosis and control group were measured and calculated by ELISA technique. Fruit: 1.Real-time PCR result: VCAM-1mRNA in ectopic uterus Compared with the control group, there were significant differences between the control group and the control group (P0.01, P0.01), but there was no statistical significance (P0.05) between the eutopic and control group (P0.05).ICAM-1mRNA in the ectopic endometrium group and the eutopic endometrium group respectively, the control group had significant difference (P0.01, P0.01), but in the control group, there was a statistically significant difference between the control group and the control group. There was no significant difference (P0.05).2.Western blot results in the endometrium group with the control group (P0.05): VCAM-1 was significantly different in the ectopic endometrium group, in the eutopic endometrium group and the control group, with statistical significance (P0.01). The order of the expression was as follows: the eutopic endometrium group control group.ICAM-1 was in the ectopic uterus. There was significant difference between the control group and the control group, and the control group was statistically significant (P0.01, P0.01), while the relative expression of ICAM-1 in the eutopic endometrium group and the control group was not statistically significant (P0.05).3.ELISA results: the average concentration of sVCAM-1 and sICAM-1 in the serum of EMs patients was calculated according to the OD value and the standard curve, and the control was compared with the control group. The difference was significantly higher, the difference was statistically significant (P0.01, P0.01).SICAM-1 concentration in stage III, stage I and stage II, the difference was significant (P0.01), the difference was statistically significant (P0.01) in the early stage I, the serum concentration of stage II patients was higher than that of the control group (P0.01), the difference was statistically significant. The ROC curve analysis, serum sICAM-1 and s were used. VCAM-1 has a certain accuracy in the diagnosis of endometriosis. Among them, sICAM-1 has high specificity and low sensitivity. Conclusion: 1.VCAM-1 and ICAM-1 can participate in the occurrence and development of endometriosis through the level of mRNA in patients with endometriosis. There is no significant correlation between the expression of.2.VCAM-1 and ICAM-1 in the endometriosis of the patients with endometriosis and the expression of the eutopic tissue protein in endometriosis, suggesting that the pathogenesis of endometriosis may be the different.3. peripheral blood sICAM-1 and sVCAM-1 in the diagnosis of endometriosis patients. There is a certain degree of accuracy. Detection of serum sICMA-1 concentration can help to understand the progress of endometriosis and may become a new diagnostic method for endometriosis.

【学位授予单位】：浙江大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R711.71

【参考文献】