卵巢功能衰退的比较蛋白质组学及相关蛋白差异表达的研究

发布时间：2018-02-13 08:46

本文关键词： 蛋白质组学卵巢蛋白表达 GLO I UCH-L1 HSP27 SAP　出处：《吉林大学》2014年博士论文　论文类型：学位论文

【摘要】：卵巢功能提前衰退，严重影响女性生殖健康及生活质量。因此关于卵泡发育成熟及卵巢功能衰退方面的研究一直是关注的热点。本研究取绝经期与育龄期女性卵巢组织进行比较蛋白质组学分析，也是对卵巢功能衰退及卵巢功能正常的卵巢组织进行比较分析，提取两组卵巢组织蛋白质，应用荧光差异凝胶电泳(difference gel electrophoresis，DIGE)及MALDI-TOF质谱技术筛选出功能蛋白，通过western blot技术行蛋白差异表达分析、免疫组化技术行蛋白定位表达分析。功能衰退与功能正常卵巢组织中存在差异表达蛋白点31个，其中表达上调14个，表达下调17个。存在4个差异表达蛋白质乙二醛酶I(Glyoxalase I,GLO I)、泛素羧基末端水解酶L1(Ubiquitin carboxy-terminal hydrolase L1，，UCH-L1)、热休克蛋白27(heat shock protein27，HSP27）、血清淀粉样蛋白P(serum amyloid P，SAP)。将这4种蛋白作为功能蛋白，它们在两组人群卵巢组织中均有表达，在功能衰退卵巢组织中表达升高。且4种蛋白均在功能衰退卵巢闭锁卵泡中表达增强；并在不同发育阶段卵泡细胞中表达水平强弱不一。SAP在不同发育阶段卵泡细胞中均有较强表达；HSP27在原始卵泡、初级卵泡卵母细胞胞浆及闭锁卵泡表达增强；GLO I在成熟卵泡颗粒细胞及卵泡膜细胞胞浆中表达增强，在闭锁卵泡中表达最强；UCH-L1在成熟卵泡颗粒细胞及卵泡膜细胞胞浆中呈强表达，闭锁卵泡中表达较生长卵泡细胞中减弱；4种蛋白在卵巢白体中均不表达。本实验首次应用比较蛋白质组学技术分析人类育龄期与绝经期女性卵巢组织的蛋白差异表达。4种差异表达蛋白可以作为判断卵巢功能的指标，作为调控卵泡发育、预防及诊治卵巢早衰及延缓卵巢功能衰退的理想研究靶点。
[Abstract]:Premature decline in ovarian function, Therefore, the research on follicular maturation and ovarian function decline has been a hot topic. The comparative proteomic analysis of ovarian tissues in menopausal and childbearing age was carried out in this study. The functional proteins of the two groups of ovarian tissues were extracted by fluorescence differential gel electrophoresis (FGE) and differential gel electrophoresistics (DIGE) and MALDI-TOF mass spectrometry. The differential expression of proteins was analyzed by western blot technique, and the localization of proteins was analyzed by immunohistochemical technique. There were 31 differentially expressed proteins in functional decline and normal ovarian tissues, of which 14 were up-regulated. There were 4 differentially expressed proteins Glyoxalase I (Glyoxalase I), Ubiquitin carboxy-terminal carboxy-terminal hydrolase L1 (UCH-L1), heat shock protein (HSP27) and amyloid protein (amyloid) as functional proteins. They were expressed in ovarian tissues of both groups, but increased in functional decline ovarian tissues, and the expression of four proteins was enhanced in atresia follicles. SAP had strong expression of HSP27 in follicular cells at different stages of development, and HSP27 was expressed in primordial follicles. The expression of GLO I in the cytoplasm of primary follicular oocytes and atresia follicles was enhanced in granulosa cells and theca cells of mature follicles. The strongest expression of UCH-L1 in atresia follicles was observed in granulosa cells and theca cells of mature follicles, but the expression of four proteins in atretic follicles was less than that in cultured follicles. For the first time, comparative proteomics was used to analyze the differential expression of 4 proteins in ovarian tissues of women at reproductive and menopausal stages, which can be used as an index to judge ovarian function and to regulate follicle development. Ideal targets for prevention, diagnosis and treatment of premature ovarian failure and delay of ovarian dysfunction.
【学位授予单位】：吉林大学
【学位级别】：博士
【学位授予年份】：2014
【分类号】：R711.75

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