發(fā)布時間：2018-02-13 08:46

  本文關(guān)鍵詞： 蛋白質(zhì)組學(xué) 卵巢 蛋白表達(dá) GLO I UCH-L1 HSP27 SAP　出處：《吉林大學(xué)》2014年博士論文　論文類型：學(xué)位論文

【摘要】：卵巢功能提前衰退，嚴(yán)重影響女性生殖健康及生活質(zhì)量。因此關(guān)于卵泡發(fā)育成熟及卵巢功能衰退方面的研究一直是關(guān)注的熱點。本研究取絕經(jīng)期與育齡期女性卵巢組織進(jìn)行比較蛋白質(zhì)組學(xué)分析，也是對卵巢功能衰退及卵巢功能正常的卵巢組織進(jìn)行比較分析，提取兩組卵巢組織蛋白質(zhì)，應(yīng)用熒光差異凝膠電泳(difference gel electrophoresis，DIGE)及MALDI-TOF質(zhì)譜技術(shù)篩選出功能蛋白，通過western blot技術(shù)行蛋白差異表達(dá)分析、免疫組化技術(shù)行蛋白定位表達(dá)分析。 功能衰退與功能正常卵巢組織中存在差異表達(dá)蛋白點31個，其中表達(dá)上調(diào)14個，表達(dá)下調(diào)17個。存在4個差異表達(dá)蛋白質(zhì)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)、熱休克蛋白27(heat shock protein27，HSP27）、血清淀粉樣蛋白P(serum amyloid P，SAP)。將這4種蛋白作為功能蛋白，它們在兩組人群卵巢組織中均有表達(dá)，在功能衰退卵巢組織中表達(dá)升高。且4種蛋白均在功能衰退卵巢閉鎖卵泡中表達(dá)增強(qiáng)；并在不同發(fā)育階段卵泡細(xì)胞中表達(dá)水平強(qiáng)弱不一。SAP在不同發(fā)育階段卵泡細(xì)胞中均有較強(qiáng)表達(dá)；HSP27在原始卵泡、初級卵泡卵母細(xì)胞胞漿及閉鎖卵泡表達(dá)增強(qiáng)；GLO I在成熟卵泡顆粒細(xì)胞及卵泡膜細(xì)胞胞漿中表達(dá)增強(qiáng)，在閉鎖卵泡中表達(dá)最強(qiáng)；UCH-L1在成熟卵泡顆粒細(xì)胞及卵泡膜細(xì)胞胞漿中呈強(qiáng)表達(dá)，閉鎖卵泡中表達(dá)較生長卵泡細(xì)胞中減弱；4種蛋白在卵巢白體中均不表達(dá)。 本實驗首次應(yīng)用比較蛋白質(zhì)組學(xué)技術(shù)分析人類育齡期與絕經(jīng)期女性卵巢組織的蛋白差異表達(dá)。4種差異表達(dá)蛋白可以作為判斷卵巢功能的指標(biāo)，作為調(diào)控卵泡發(fā)育、預(yù)防及診治卵巢早衰及延緩卵巢功能衰退的理想研究靶點。
[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.
【學(xué)位授予單位】：吉林大學(xué)
【學(xué)位級別】：博士
【學(xué)位授予年份】：2014
【分類號】：R711.75

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本文編號：1507766