邻苯二甲酸酯暴露与稽留流产相关性及机制研究

发布时间：2018-03-02 07:39

本文关键词： 稽留流产邻苯二甲酸酯凋亡 Bax Bcl-2　出处：《复旦大学》2014年硕士论文　论文类型：学位论文

【摘要】：[目的]通过临床资料统计与实验研究相结合的方法,揭示邻苯二甲酸酯(PAEs)与稽留流产的关系,并初步探讨其发病机制。[方法](1)通过病例-对照回顾性研究,收集1993年-2012年我院稽留流产病例,共计1945例,对比1874例妊娠20周内的单胎活孕妊娠者的病例资料。通过卡方检验和logistic回归分析,分析与稽留流产发病相关的高危因素,探讨环境等因素在稽留流产中可能的影响。(2)研究稽留流产病人中邻苯二甲酸酯(PAEs)的暴露情况：留取原因不明的稽留流产病人及正常妊娠者各150份同期尿液标本。采用超高效液相色谱-质谱法分别检测尿液中13种邻苯二甲酸酯代谢产物水平,研究稽留流产病人的邻苯二甲酸酯(PAEs)暴露情况。(3)体外试验：观察人滋养层细胞系(HTR-8)在不同浓度DEHP (PAEs最常见的一种)作用下细胞活力、凋亡及细胞周期变化情况,初步探讨其凋亡相关机制。[结果](1)本案logistic回归多因素分析筛选出5个独立危险因素,将其按OR值由大至小进行排列如下：乙型肝炎病毒携带者(OR 8.126,CI4.688-14.084),贫血(OR 5.498,CI 2.243-13.473),初产妇(OR 2.305,CI 2.013-2.638),自然流产史(OR 2.359,CI 1.703-3.268),室外职业者(OR1.777,CI 1.538-2.052)。研究近年来稽留流产独立因素构成变化显示：室外职业及环境污染构成比升高(P0.05)与稽留流产发病数增加有关；而流产≥3次、自然流产病史、贫血和乙肝病毒携带者的构成比变化与稽留流产无明显相关(P0.05)；同样,初产妇这一稽留流产危险因素,占比呈明显下降与稽留流产发病数升高不符合。(2)超高效液相色谱-质谱法检测结果：DEHP的三种代谢产物(MEHP、MEHHP、MEOHP)及DMP代谢产物MMP在病例组尿液中检出水平显著高于对照组(P0.05)。而DEP的代谢产物MEP在病例组尿液中检出水平却显著低于对照组(P0.05)。在相关性分析中,MEHP与雌激素中度相关,而MMP与孕激素中度相关。(3)体外研究发现：①DEHP≤20ng/ml作用于HTR-8细胞48h未发现明显毒性作用；DEHP 15ng/ml作用24小时能抑制HTR-8细胞的凋亡,同时明显延长细胞S期,减少GO/Gl期阻滞。DEHP 15ng/ml作用HTR-8细胞系6小时,发现Bax表达降低而Bcl-2表达上升,可能是小剂量DEHP抑制细胞凋亡的机制。②DEHP≥30ng/ml作用HTR-8细胞24到48小时,HTR-8细胞活性明显受抑制(P0.05); 30ng/ml的DEHP能促进HTR-8细胞凋亡,同时显著性缩短细胞S期,增加GO/G1期阻滞。[结论](1)室外职业者、初产妇、自然流产病史、贫血和乙肝病毒携带者可能是稽留流产发生的独立危险因素,其中仅有室外职业者及未知因素的构成比随稽留流产发病数增加而升高。初步推论生育史、合并症不能解释稽留流产发病数的升高,而环境污染等不明原因可能是稽留流产病例数升高的重要原因之一。(2)稽留流产病人部分邻苯二甲酸酯暴露情况明显高于对照组,表明环境污染物-邻苯二甲酸酯与近年来稽留流产高发病数有关。(3) DEHP 15ng/ml对人早孕滋养细胞毒性不明显,并能抑制其凋亡,促进生长。其原因可能与小剂量的DEHP能下调Bax表达,上调Bcl-2的表达有关,是稽留流产发生的机制之一。
[Abstract]:[Objective] by the combination of statistical methods and experimental study of the clinical data, reveals the adjacent benzene two formic acid ester (PAEs) and the relationship between missed abortion, and to explore its pathogenesis. Methods: (1) through retrospective case-control study, collected in our hospital in 1993 -2012 missed abortion patients, a total of 1945 cases and compared with 1874 cases of singleton pregnancy within 20 weeks of pregnancy who live cases. Through chi square test and logistic regression analysis, analysis of risk factors associated with the incidence of missed abortion, to explore the impact of environmental factors may in missed abortion. (2) to study the adjacent benzene two formic acid ester with abortion patients (PAEs) exposure: take2ml of unexplained abortion patients and normal pregnant women during the same period. 150 copies of urine samples in the urine of 13 kinds of phthalate ester two formic acid metabolites were detected by ultra performance liquid chromatography mass spectrometry. The research of missed abortion patients Two people of phthalic acid ester (PAEs) exposure. (3) in vitro: Observation of human trophoblast cell line (HTR-8) in different concentrations of DEHP (one of the most common PAEs) cell viability under the effect of changes in cell cycle and apoptosis, to explore the mechanism of apoptosis related results (1)]. In the case of multiple logistic regression analysis identified 5 independent risk factors, according to the OR value from large to small order as follows: hepatitis B virus (OR 8.126, CI4.688-14.084), anemia (OR 5.498, CI 2.243-13.473), primipara (OR 2.305, CI 2.013-2.638), the history of spontaneous abortion (OR 2.359 CI, 1.703-3.268), outdoor occupation (OR1.777, CI 1.538-2.052). The independent risk factors of missed abortion in recent years showed that: the proportion of changes in environmental pollution and increase of outdoor occupation (P0.05) is associated with increased incidence of missed abortion and abortion; more than 3 times, abortion history, and anemia A hepatitis B virus carriers than changes and missed abortion and no obvious correlation (P0.05); similarly, the early maternal risk factors of missed abortion, the proportion decreased significantly with increased incidence of missed abortion is not met. (2) the detection results of ultra high performance liquid chromatography mass spectrometry: three metabolites DEHP (MEHP MEHHP, MEOHP, DMP) and metabolites of MMP detected in the case group in the urine was significantly higher than the control group (P0.05) and DEP. The metabolites of MEP in case group in the urine detection level was significantly lower than that of control group (P0.05). In correlation analysis, MEHP and estrogen and progesterone and MMP are moderate, moderate correlation. (3) in vitro was as follows: DEHP = 20ng/ml 48h in HTR-8 cells without obvious toxicity; 15ng/ml can inhibit the apoptosis of DEHP HTR-8 cells for 24 hours, at the same time prolong cell S phase, GO/Gl phase arrest of.DEHP 15ng/ml reduction Using HTR-8 cell lines for 6 hours showed that the expression of Bax decreased and Bcl-2 expression increased, is a possible mechanism of low dose of DEHP inhibits apoptosis. The DEHP 30ng/ml or HTR-8 cells 24 to 48 hours, the activity of HTR-8 cells was significantly inhibited (P0.05); 30ng/ml DEHP can promote the apoptosis of HTR-8 cells, and significantly reduce cell S phase, GO/G1 phase arrest increased. Conclusion: (1) outdoor occupation, primipara, spontaneous abortion history, anemia and hepatitis B virus carriers may be independent risk factors of missed abortion, which constitute the only outdoor occupation and unknown factors than with missed abortion incidence increased. We infer that reproductive history. Complications can not explain the increased incidence of missed abortion, and environmental pollution and other unknown reasons may be one of the important reasons of missed abortion were increased. The number of missed abortion patients (2) part of the adjacent benzene two phthalate exposure Significantly higher than the control group, indicating that the environmental pollutants - phthalate two formic acid ester and in recent years, the high incidence of missed abortion number. (3) DEHP 15ng/ml on trophoblast cell toxicity was not obvious, and can inhibit apoptosis and promote growth. The reason may be related to small doses of DEHP can downregulate Bax expression and up-regulated expression Bcl-2, is one of the mechanisms of missed abortion.

【学位授予单位】：复旦大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R714.21

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