發(fā)布時(shí)間：2018-06-04 19:47

  本文選題：DEHP + 胚胎著床��； 參考：《重慶醫(yī)科大學(xué)》2012年碩士論文

【摘要】：DEHP作為增塑劑及化妝品添加劑在生活中被廣泛接觸，目前已在全球形成嚴(yán)重的環(huán)境污染和人類暴露。大量研究結(jié)果顯示DEHP嚴(yán)重危害機(jī)體健康，特別是產(chǎn)生生殖毒性，如導(dǎo)致雄性隱睪和精子發(fā)生障礙。事實(shí)上，女性由于長(zhǎng)期接觸化妝品及洗滌用品比男性暴露于DEHP的幾率更高，已有數(shù)據(jù)顯示雌性DEHP暴露與不良妊娠密切相關(guān)，但是DEHP導(dǎo)致不良妊娠的原因尚不明確。胚胎植入作為整個(gè)妊娠過程中關(guān)鍵環(huán)節(jié)，其成功與否直接影響妊娠結(jié)局。成功的胚胎植入依賴受到子宮內(nèi)膜容受性、胚胎活力、絨毛外滋養(yǎng)層細(xì)胞侵襲能力等多因素影響。那么DEHP暴露是否干擾了胚胎植入過程中的這些因素呢？ 為了探明DEHP對(duì)胚胎著床的影響及其相關(guān)干擾途徑，，本研究主要進(jìn)行了以下實(shí)驗(yàn)。首先利用孕鼠作為研究對(duì)象，探討DEHP對(duì)胚胎著床的影響及其分子機(jī)制，主要實(shí)驗(yàn)內(nèi)容包括：給予交配后的雌鼠不同劑量DEHP處理，收集材料，計(jì)數(shù)胚胎著床數(shù)目；利用HE染色與掃描電鏡觀察子宮組織形態(tài)學(xué)，檢測(cè)DEHP對(duì)子宮內(nèi)膜容受性主要形態(tài)標(biāo)識(shí)的影響；采用real-timePCR,westernblot和免疫組化分別檢測(cè)DEHP對(duì)子宮內(nèi)膜容受性相關(guān)分子ERα,PR,E-cadherin,HoxA10,MMP-2及其調(diào)控因子表達(dá)的影響。其次，以絨毛外滋養(yǎng)層細(xì)胞HTR-8/Svneo為模型，探討DEHP對(duì)其侵襲和遷移能力的影響，主要實(shí)驗(yàn)包括：用MTT法和流式細(xì)胞術(shù)確定DEHP的作用濃度；Transwell小室檢測(cè)DEHP對(duì)細(xì)胞侵襲和遷移能力的影響，Real-timePCR和Westernblot檢測(cè)侵襲與遷移相關(guān)因子表達(dá)的變化。上述研究發(fā)現(xiàn)： 1.DEHP暴露可損傷子宮內(nèi)膜容受性建立并降低胚胎著床率，進(jìn)而引發(fā)不良妊娠結(jié)局。 2.DEHP暴露可通過上調(diào)ERα,PR及E-cadherin等容受性相關(guān)因子分子表達(dá)破壞子宮內(nèi)膜容受態(tài)的平衡，從而影響胚胎著床。 3.DEHP對(duì)小鼠子宮內(nèi)膜組織中E-cadherin的表達(dá)調(diào)控受MAPK與NF-κB信號(hào)通路影響。 4.DEHP可通過上調(diào)MMP-2與MMP-9的表達(dá)顯著促進(jìn)絨毛外滋養(yǎng)層細(xì)胞HTR-8/Svneo的侵襲和遷移，干擾絨毛外滋養(yǎng)層細(xì)胞對(duì)子宮內(nèi)膜組織的適度侵襲，導(dǎo)致其過度或失控侵襲，并有可能引發(fā)妊娠相關(guān)的滋養(yǎng)細(xì)胞腫瘤發(fā)生 5.DEHP上調(diào)MMP-2與MMP-9的表達(dá)受p38、JNK的磷酸化的調(diào)控。 綜上可知，DEHP可通過損傷小鼠子宮內(nèi)膜容受性并干擾絨毛外滋養(yǎng)層細(xì)胞的適度侵襲導(dǎo)致胚胎著床異常，進(jìn)而造成不良妊娠結(jié)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 偽 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.
【學(xué)位授予單位】：重慶醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2012
【分類號(hào)】：R321

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