增塑剂DEHP对胚胎着床的影响及其分子机制初探

发布时间：2018-06-04 19:47

本文选题：DEHP + 胚胎着床　；参考：《重庆医科大学》2012年硕士论文

【摘要】：DEHP作为增塑剂及化妆品添加剂在生活中被广泛接触，目前已在全球形成严重的环境污染和人类暴露。大量研究结果显示DEHP严重危害机体健康，特别是产生生殖毒性，如导致雄性隐睾和精子发生障碍。事实上，女性由于长期接触化妆品及洗涤用品比男性暴露于DEHP的几率更高，已有数据显示雌性DEHP暴露与不良妊娠密切相关，但是DEHP导致不良妊娠的原因尚不明确。胚胎植入作为整个妊娠过程中关键环节，其成功与否直接影响妊娠结局。成功的胚胎植入依赖受到子宫内膜容受性、胚胎活力、绒毛外滋养层细胞侵袭能力等多因素影响。那么DEHP暴露是否干扰了胚胎植入过程中的这些因素呢？为了探明DEHP对胚胎着床的影响及其相关干扰途径，，本研究主要进行了以下实验。首先利用孕鼠作为研究对象，探讨DEHP对胚胎着床的影响及其分子机制，主要实验内容包括：给予交配后的雌鼠不同剂量DEHP处理，收集材料，计数胚胎着床数目；利用HE染色与扫描电镜观察子宫组织形态学，检测DEHP对子宫内膜容受性主要形态标识的影响；采用real-timePCR,westernblot和免疫组化分别检测DEHP对子宫内膜容受性相关分子ERα,PR,E-cadherin,HoxA10,MMP-2及其调控因子表达的影响。其次，以绒毛外滋养层细胞HTR-8/Svneo为模型，探讨DEHP对其侵袭和迁移能力的影响，主要实验包括：用MTT法和流式细胞术确定DEHP的作用浓度；Transwell小室检测DEHP对细胞侵袭和迁移能力的影响，Real-timePCR和Westernblot检测侵袭与迁移相关因子表达的变化。上述研究发现： 1.DEHP暴露可损伤子宫内膜容受性建立并降低胚胎着床率，进而引发不良妊娠结局。 2.DEHP暴露可通过上调ERα,PR及E-cadherin等容受性相关因子分子表达破坏子宫内膜容受态的平衡，从而影响胚胎着床。 3.DEHP对小鼠子宫内膜组织中E-cadherin的表达调控受MAPK与NF-κB信号通路影响。 4.DEHP可通过上调MMP-2与MMP-9的表达显著促进绒毛外滋养层细胞HTR-8/Svneo的侵袭和迁移，干扰绒毛外滋养层细胞对子宫内膜组织的适度侵袭，导致其过度或失控侵袭，并有可能引发妊娠相关的滋养细胞肿瘤发生 5.DEHP上调MMP-2与MMP-9的表达受p38、JNK的磷酸化的调控。综上可知，DEHP可通过损伤小鼠子宫内膜容受性并干扰绒毛外滋养层细胞的适度侵袭导致胚胎着床异常，进而造成不良妊娠结局。
[Abstract]:DEHP, as a plasticizer and cosmetic additive, has been widely contacted in daily life, and has been seriously polluted and exposed to human beings all over the world. A large number of studies have shown that DEHP is a serious health hazard, especially reproductive toxicity, such as male cryptorchidism and spermatogenesis disorders. In fact, women are more likely to be exposed to DEHP than men because of long-term exposure to cosmetics and detergents. Data have shown that female DEHP exposure is closely related to poor pregnancy, but the cause of DEHP is unclear. Embryo implantation is a key step in the whole process of pregnancy, and its success directly affects the outcome of pregnancy. Successful embryo implantation dependence was influenced by endometrial receptivity, embryo viability and invasion ability of extravillous trophoblast cells. So does DEHP exposure interfere with these factors in the implantation process? In order to investigate the effect of DEHP on embryo implantation and its related interference pathway, the following experiments were carried out in this study. Firstly, the effects of DEHP on embryo implantation and its molecular mechanism were studied by using pregnant mice as the research object. The main contents of the experiments were as follows: the female mice after mating were treated with different doses of DEHP, collected materials and counted the number of embryo implantation; The morphology of uterus was observed by HE staining and scanning electron microscope (SEM). The effect of DEHP on the main morphological markers of endometrial receptivity was detected. The effects of DEHP on the expression of MMP-2 and its regulatory factors were detected by real-time PCR Western blot and immunohistochemistry, respectively. Secondly, the effects of DEHP on the invasion and migration of trophoblastic cells (HTR-8/Svneo) were studied. The main results were as follows: MTT assay and flow cytometry were used to determine the concentration of DEHP. Transwell chamber was used to detect the effect of DEHP on the ability of cell invasion and migration. Real-time PCR and Westernblot were used to detect the expression of invasion and migration related factors. The study found that: 1.DEHP exposure can damage endometrial receptivity and reduce embryo implantation rate, leading to adverse pregnancy outcomes. 2.DEHP exposure may affect embryo implantation by up-regulating the expression of reception-related factors such as ER 伪 PR and E-cadherin. The regulation of E-cadherin expression in mouse endometrium by 3.DEHP is affected by MAPK and NF- 魏 B signaling pathway. By upregulating the expression of MMP-2 and MMP-9, 4.DEHP could significantly promote the invasion and migration of extracellular trophoblast cells (HTR-8/Svneo), and interfere with the moderate invasion of extravillous trophoblast cells to endometrium, leading to excessive or uncontrolled invasion. And may lead to gestational gestation-related trophoblastic neoplasms. 5.DEHP upregulated the expression of MMP-2 and MMP-9 by the phosphorylation of p38 JNK. It can be concluded that DEHP can result in abnormal implantation of embryo by damaging the endometrial receptivity of mice and interfering with the moderate invasion of extravillous trophoblastic cells, which leads to the adverse pregnancy outcome.
【学位授予单位】：重庆医科大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R321

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