【作者】 刘欣燕；

【导师】 边旭明；

【作者基本信息】 中国协和医科大学 ， 妇产科学， 2007， 博士

【摘要】 研究背景:早期自然流产是一种严重影响妇女身心健康的疾病,目前尚缺乏有效的诊断和治疗的办法。其发病原因非常复杂,涉及妇产科学、遗传学、流行病学、职业医学、免疫学、血液及内分泌学等多个学科。仅仅局限在一个狭窄领域的病因学研究,难以获得对早期自然流产的全方位认识。目前国内外尚缺乏从不同角度来阐述早期自然流产发病原因的报道。随着科技的发展,大气污染、水污染、噪声污染和电磁辐射污染已经成为人类生活环境中的四种主要污染源,但到目前为止,生活环境中的有害因素对妊娠的影响尚未得到足够的重视,没有相关的基本流行病学资料。虽然人们早就认识到胚胎的细胞遗传学异常是早期自然流产的重要原因之一,但引起其异常的原因目前仍然不清楚。现阶段的蛋白质研究多停留在是研究已知的蛋白质与早期自然流产关系的水平上,所能涉及的蛋白质数量非常有限。近年来迅速发展的蛋白质组学技术,高通量、高灵敏度和规模化的双向凝胶电泳结合质谱技术为我们提供了一个能够比较全面认识组织蛋白质成份的平台,可以帮助我们寻找早期自然流产特异表达的蛋白质。如果能将这些特异表达的蛋白质作为临床诊断的生物标记分子或药物开发的分子靶标,早期自然流产就有望成为可以早期诊断并治疗的疾病。研究目的:比较自然流产患者和正常妊娠妇女流行病学特点和蛋白质表达的差异、分析自然流产患者的绒毛染色体核型,从流行病学、蛋白质组学和细胞遗传学三个角度探讨自然流产的发病原因。研究方法:1.流行病学调查。问卷调查200对自然流产患者和正常足月分娩妇女在早孕期的生活环境情况,分析环境因素对妊娠的影响。2.细胞遗传学研究。分析453例早期自然流产患者的绒毛染色体核型,研究早期自然流产的细胞遗传学特点。3.蛋白质组学研究。采用双向凝胶电泳结合MALDI质谱分析技术,比较自然流产患者和正常妊娠妇女的绒毛组织中蛋白质表达的差异,寻找自然流产特异表达的蛋白质。结果:1.单因素分析显示自然流产组平均每周看电视在10小时以上,使用电脑在45小时以上、经常使用复印机、微波炉和手机的比例显著高于对照组(P＜0.05);自然流产组生活环境附近有高压线等电磁辐射装置、有排放污染气体的工厂、有持续10个月以上的噪音、孕期或孕前半年家庭或工作场所采用挥发异味的材料进行过装修的比例也显著高于对照组(P＜0.05),且随着装修异味时间的延长,发生自然流产的危险度增加(P＜0.001)。自然流产组孕期情绪紧张的比例显著高于对照组(P＜0.001),有自然流产史的比例也显著高于对照组(P＜0.001)。多因素分析显示早孕期经常使用微波炉、手机、接触装修材料散发的异味的时间超过3个月、情绪紧张以及有自然流产史等5种因素显著增加自然流产的相对危险性(P＜0.05)。2.自然流产患者绒毛染色体核型异常率为58.04%;多次流产患者的绒毛染色体核型异常率明显高于初次流产患者(P＜0.05);绒毛染色体核型的异常绝大多数为染色体数目异常,其中16—三体最多见;早孕期生活环境中的高危因素(如:早孕期经常使用微波炉、手机、接触装修材料异味超过3个月、情绪紧张)并不增加绒毛染色体核型异常的比率。3.在自然流产患者绒毛组织及正常妊娠的绒毛组织中共鉴定出14种差异表达的蛋白质,其中8种蛋白质在自然流产患者绒毛组织中表达下调,6种蛋白质在自然流产患者绒毛组织中表达上调。谷胱甘肽S-转移酶、氯离子细胞内通道3、雌激素17β-羟基类固醇脱氢酶、膜联蛋白Ⅰ、羟基前列腺素脱氢酶15(NAD)、组织蛋白酶和核糖体这7种蛋白质在以往的报告中与胚胎的发育有关。谷胱甘肽S-转移酶、氯离子细胞内通道3在不同组间重复出现,推测可能与自然流产有关。结论:1.早孕期经常使用微波炉、手机、接触装修材料散发的异味的时间超过3个月、情绪紧张和有自然流产史是早期自然流产的高危因素。2.染色体核型异常特别是染色体数目异常是早期自然流产的重要原因,在复发性流产中更重要。早孕期生活环境中的高危因素对绒毛染色体核型没有明显影响。3.双向凝胶电泳结合MALDI质谱分析这两项蛋白质组学技术可以用于分析早孕绒毛标本中的差异表达蛋白质。对这些标记蛋白质的进一步研究,可以为早期自然流产的发病机制、早期诊断和治疗提供重要的信息。更多还原

【Abstract】 BackgroundEarly spontaneous abortion is a devastating reproductive problem affecting mental and physical health of women. Many researches have sought to determine its causation and just a handful factors have been found to be associated with this disorder. The pathogenesis of spontaneous abortion is complicated, and the etiology of spontaneous abortion involves in several subjects, such as Obstetrics & Gynecology, Genetics, Epidemiology, Immunology, Hematology and Endocrinology and so on. A study limited in a narrow area will be very difficult to gain valuable information about this disease.ObjectiveTo investigate the etiological factors of early spontaneous abortion form the view of Epidemiology, Genetics and Proteomics, we compared the epidemiological characteristics and the differences of the express of the proteins between women undergone spontaneous abortion and who didn’t undergone spontaneous abortion ; and analysis the embryonic chorionic karyotypes of abortion.Research Method1. Epidemiologic survey. To conduct analysis based on the survey of 200 early spontaneous abortion cases and the matched control group about the state of living environment by using multifactor Logistic regression analysis.2. Study of Cytogenetics. Examine 453 chorionic karyotypes in early spontaneous abortion, to research the cytogenetical characteristics of this disorder.3. Study of Proteomics. Using 2-DE combined with MALDI-TOF mass spectrum to identify the differential expressed proteins: 6 cases of early spontaneous abortion and the matched normal control were analysis by this approach. Results1. Five factories in living environment of first trimester were associated with risk of early spontaneous abortion significantly by multifactor analysis. They are using microwave oven and mobile phone usually, the duration of contacting abnormal smell of fitment material was more than 3 months, having emotional stress during the first trimester of pregnancy and having early spontaneous abortion history (P<0.05) .2. There hundred thirty nine abortuses were successfully karyityped, 58.04% had chromosome aberrations. The abnormal rate of chorionic karyotype in cases of repeat early spontaneous abortion was significantly higher than that in first early spontaneous abortion cases (P<0.05) . Most of abnormal karyotypes were numerical abnormality. Some risk factories (such as using microwave oven and mobile phone usually, the duration of contacting abnormal smell of fitment material was more than 3 months, having emotional stress during the first trimester of pregnancy) in living environment during the first trimester weren’t associated with chorionic karyotypes.3. Fourteen differential expressed proteins were identified by 2-DE combined MALDI-TOF mass spectrum. Eight of these proteins showed decreased expression in chorionic tissue of early spontaneous abortion and six of these proteins showed increased expression in chorionic tissue of early spontaneous abortion. Of these differentially expressed proteins, seven were previously reported to be associated with embryo development.Conclusion1. Usually use microwave oven and mobile phone , contacting abnormal smell of fitment material more than 3 months, having emotional stress during the first trimester of pregnancy and having early spontaneous abortion history were risk factories for early spontaneous abortion .2. Abnormality of the chromosomal karyotype especially the abnormality of the number of chromosome is the most important reasons in early spontaneous abortion, especially in early repeat spontaneous abortion. Chorionic karyotype wasn’t influenced by some risk factories in living environment during the first trimester of pregnancy.3. 2-DE combined with MALDI-TOF mass spectrum was improved to be able to identify the differential expression proteins in early spontaneous abortion sample. These proteins may be serving as an additional parameter for a better insight into the study of the diagnosis and treatment of early spontaneous abortion.更多还原