本文關(guān)鍵詞：Aquaproin-8在卵泡發(fā)育中的作用與機(jī)制和阿爾茲海默癥小鼠模型血液生物標(biāo)志物研究　出處：《吉林大學(xué)》2017年碩士論文　論文類(lèi)型：學(xué)位論文

  更多相關(guān)文章： AQP8 增殖 遷移 3D 卵泡培養(yǎng) 阿爾茲海默癥 水迷宮 Aβ蛋白α-2M CFH CLUSTERIN

【摘要】：第一部分:Aquaporin-8在卵泡發(fā)育中的作用和機(jī)制研究水通道蛋白家族(aquaporins,AQPs)是一類(lèi)在細(xì)胞膜上轉(zhuǎn)運(yùn)水分子的通道蛋白,在機(jī)體液體代謝中具有重要的功能。AQP8作為家族中的一員,已被證明在消化生理、肝膽生理和生殖生理中等都發(fā)揮著作用。近年來(lái),AQP8在哺乳動(dòng)物雌性生殖系統(tǒng)中的重要性正在被逐步揭示。本課題組曾揭示AQP8在卵巢中專(zhuān)一性地表達(dá)在顆粒細(xì)胞上,AQP8的缺失導(dǎo)致小鼠顆粒細(xì)胞水通透性降低,由此引發(fā)的生理效應(yīng)是排卵數(shù)目增多和產(chǎn)仔數(shù)量增加,以及還出現(xiàn)了多卵卵泡現(xiàn)象。然而AQP8在卵泡發(fā)育中的作用和機(jī)制很大程度上還沒(méi)被人們理解。在當(dāng)前的研究里,利用AQP8基因敲除小鼠,我們分別從卵巢、卵泡、顆粒細(xì)胞三個(gè)逐漸深入的水平開(kāi)展實(shí)驗(yàn),發(fā)現(xiàn)了AQP8在卵泡發(fā)育尤其是卵泡腔形成中的新功能并闡明了相關(guān)的機(jī)理。首先我們發(fā)現(xiàn)AQP8敲除鼠相比于野生鼠有明顯增多的有腔卵泡現(xiàn)象,并且通過(guò)卵泡3D體外培養(yǎng)的方法,我們發(fā)AQP8敲除鼠的卵泡成活率和成腔率都要明顯高于野生鼠。對(duì)這一現(xiàn)象進(jìn)行進(jìn)一步分析,將野生鼠和AQP8基因敲除鼠的卵泡顆粒細(xì)胞分離、培養(yǎng)。在增殖實(shí)驗(yàn)中,FSH存在下的AQP8敲除型顆粒細(xì)胞的增殖能力相比野生型顯著減弱。在橫向與縱向遷移實(shí)驗(yàn)中,我們發(fā)現(xiàn)無(wú)論是否FSH的存在,AQP8敲除型卵泡顆粒細(xì)胞的遷移能力都要明顯弱于野生鼠的顆粒細(xì)胞。接著我們考察了顆粒細(xì)胞的雌激素和孕激素合成能力,檢測(cè)合成這兩個(gè)激素的三個(gè)限速酶CYP19A1,CYP11A1和St AR的m RNA表達(dá)水平和蛋白質(zhì)水平,發(fā)現(xiàn)兩種基因型顆粒細(xì)胞激素合成能力無(wú)區(qū)別�；谶@些實(shí)驗(yàn)結(jié)果,我們對(duì)AQP8敲除鼠有腔卵泡增多表型的機(jī)制解釋是,顆粒細(xì)胞增殖和遷移能力的減弱使卵泡在腔前期更容易形成間隙和小的液體點(diǎn),這些小的液體點(diǎn)經(jīng)過(guò)聚集形成大的卵泡腔,導(dǎo)致AQP8敲除鼠成腔率增加。本研究為卵泡發(fā)育及卵泡腔的形成理論提供了新的視點(diǎn)。第二部分:阿爾茲海默癥小鼠模型中血液生物標(biāo)志物的研究阿爾茨海默氏病(AD)作為一種漸進(jìn)性與年齡有關(guān)的神經(jīng)退行性疾病,是一個(gè)全球性的健康問(wèn)題。腦中Aβ42含量升高,形成Aβ纖維沉積和tau磷酸化等是導(dǎo)致阿爾茲海默病癥的主要原因。盡管心理測(cè)試,神經(jīng)影像學(xué)、基因檢測(cè)和腦脊液生化檢測(cè)這些手段可行,但是仍然缺乏方便、準(zhǔn)確的預(yù)測(cè)和診斷AD的血漿生物標(biāo)志物。炎性反應(yīng)已經(jīng)被證實(shí)是AD發(fā)病過(guò)程中一個(gè)很重要的過(guò)程,而很多文獻(xiàn)報(bào)道炎性因子可以作為AD發(fā)病過(guò)程中的指示因子,α-2M是一種在血漿中含量豐富的血漿蛋白,在血漿中,α巨球蛋白主要作為對(duì)抗外源多肽的防御屏障。在腦中,α-2M作用是消除淀粉樣斑塊,通過(guò)結(jié)合Aβ,防止纖維形成和增強(qiáng)Aβ的清除。CFH是一種可溶性調(diào)節(jié)因子,其首要任務(wù)是補(bǔ)體3(C3)的調(diào)節(jié),補(bǔ)體3在促炎癥反應(yīng)的刺激過(guò)程中起著核心作用。Clusterin(簇集蛋白)已被定位存在于淀粉樣斑塊中,起到抑制補(bǔ)體形成,結(jié)合Aβ,防止纖維形成,清除和運(yùn)輸Aβ跨越血腦屏障的功能。本研究旨在評(píng)價(jià)β-淀粉樣蛋白,α-2巨球蛋白(α-2M),補(bǔ)體因子H(CFH)和成簇蛋白作為血漿生物標(biāo)志物的可行性。分別用3,6,9,12月齡的APP/PS1轉(zhuǎn)基因和野生型(WT)小鼠,通過(guò)Morris水迷宮實(shí)驗(yàn)和免疫組織化學(xué)方法對(duì)小鼠認(rèn)知障礙和腦淀粉樣斑塊數(shù)目進(jìn)行評(píng)價(jià),結(jié)果顯示APP/PS1轉(zhuǎn)基因的小鼠分別在6、9和12個(gè)月呈現(xiàn)AD特征。然后我們用ELISA方法對(duì)不同AD進(jìn)展?fàn)顩r的小鼠血清中Aβ40,Aβ42,α-2M,CFH和clusterin的濃度進(jìn)行測(cè)定。結(jié)果顯示3和6月齡轉(zhuǎn)基因小鼠與對(duì)照組相比,轉(zhuǎn)基因小鼠血清Aβ42水平和Aβ42/Aβ40比值顯著增加。3和6月齡轉(zhuǎn)基因小鼠與對(duì)照組相比,血清CFH水平顯著下降。同時(shí),12個(gè)月大的轉(zhuǎn)基因小鼠與對(duì)照組相比,血清成簇蛋白水平明顯升高。在轉(zhuǎn)基因和野生型小鼠之間α-2M水平?jīng)]有明顯差異。
[Abstract]:The first part: the Aquaporin-8 study on follicular development effect and mechanism of water channel protein family (aquaporins, AQPs) is a kind of transport in the cell membrane water channel protein, the body liquid metabolism has important function of.AQP8 as a member of the family, has been shown in digestive physiology, physiology and hepatobiliary the reproductive physiology of medium played a role. In recent years, the importance of AQP8 in the female reproductive system in mammals is gradually revealed. The research group has revealed that AQP8 was specifically expressed in ovarian granulosa cells in AQP8, because of the lack of water permeability of mouse granulosa cells decreased, physiological effects which caused the increase of the number of the number of ovulation and litter increased, and also appeared in a number of egg follicles. However, phenomenon and mechanism of AQP8 on follicular development largely not understood. In the present study, the use of AQP 8 gene knockout mice, we were from the ovarian follicles, granulosa cells, three levels gradually carry out the experiment, we found AQP8 in follicular development especially the new function in the formation of follicular cavity and clarify the related mechanism. First we found that AQP8 knockout mice compared to wild-type mice have significantly increased antral follicles the phenomenon, and through the method of follicle culture 3D in vitro, we AQP8 knockout mouse follicle survival rate and cavity rate are significantly higher than the wild rats. Further analysis on this phenomenon, the wild-type mice and AQP8 knockout mouse granulosa cells cultured in isolation, proliferation. In the experiment, FSH the presence of AQP8 knockout granulosa cell proliferation was significantly reduced compared to the wild type. In the horizontal and vertical migration experiment, we found that regardless of whether the presence of FSH, AQP8 knockout migration type granulosa cells are significantly weaker than Granulosa cells in wild mice. Then we investigated the ability of granulosa cells to estrogen and progesterone synthesis, three CYP19A1 synthesis rate limiting enzyme detection of these two hormones, m RNA CYP11A1 and St AR expression and protein level, found two genotypes of granulosa cell hormone synthesis ability difference. Based on the experimental results, we the AQP8 knockout mice have the mechanism to explain antral follicle phenotype is increased, weakening of granulosa cell proliferation and migration ability of the follicles are more likely to form a small gap and the liquid in the cavity period, these small liquid after together to form large follicular cavity, resulting in AQP8 knockout mouse cavity rate increased. The research the theory provides a new perspective of follicular development and follicle cavity formation. The second part: Alzheimer's disease in a mouse model of blood biomarkers of Alzheimer's disease (AD) is a progressive and years Age related neurodegenerative disease, is a global health problem. Increased brain A beta 42 content, the formation of A beta fiber deposition and tau phosphorylation is a major cause of Alzheimer's disease. Despite the psychological test, neuroimaging, gene detection and detection of cerebrospinal fluid biochemical these means is feasible, but still the lack of convenient, plasma biomarkers predictive and accurate diagnosis of AD. Inflammatory reaction has been proved to be a very important process in the pathogenesis of AD, and many reported inflammatory factors can be used as indicator in the pathogenesis of AD, a -2M is a rich content of plasma protein in the plasma. In plasma, alpha macroglobulin as the main defense barrier against exogenous peptide. In the brain, a -2M role is to eliminate amyloid plaques, through a combination of A beta, preventing fiber formation and enhancement of A beta.CFH is a soluble regulatory clearance Its primary task is to complement factor 3 (C3) regulation, complement 3 in stimulating proinflammatory process plays a central role in.Clusterin (clusterin) has been positioned in amyloid plaques, inhibit the formation of complement, combined with the A beta, prevent fiber formation, removal and transportation across the A beta the function of the blood brain barrier. The purpose of this study was to evaluate the beta amyloid protein, alpha -2 macroglobulin (alpha -2M), complement factor H (CFH) and clusters of proteins as a plasma biomarker. The feasibility of respectively 3,6,9,12 month old APP/PS1 transgenic and wild-type (WT) mice by Morris water maze test the methods of immunohistochemistry and the number of mice with cognitive impairment and cerebral amyloid plaques were evaluated. The results showed that APP/PS1 transgenic mice showed features of AD in 6,9 and 12 months respectively. Then we use the ELISA method in serum of different AD status in A 40 beta, alpha A beta 42, -2M, C The concentration of FH and clusterin were measured. The results showed that 3 and June age transgenic mice compared with the control group, the serum of transgenic mice A beta 42 beta 40 beta 42/A level and A ratio were significantly increased in.3 and June age transgenic mice compared with the control group, the serum CFH level was significantly decreased. At the same time, the 12 month old transgenic mice compared with the control group, the serum protein levels were significantly increased. Clustering between transgenic and wild-type mice did not differ significantly between alpha -2M levels.

【學(xué)位授予單位】：吉林大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類(lèi)號(hào)】：R749.16;R-332

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6 張文捷;鵝等級(jí)前卵泡發(fā)育中的基因表達(dá)差異富集及AKR1D1基因功能初探[D];四川農(nóng)業(yè)大學(xué);2016年

7 羅霏菲;黃體期高低飼喂水平影響湖羊卵泡發(fā)育的表達(dá)譜分析[D];南京農(nóng)業(yè)大學(xué);2014年

8 王德江;Aquaproin-8在卵泡發(fā)育中的作用與機(jī)制和阿爾茲海默癥小鼠模型血液生物標(biāo)志物研究[D];吉林大學(xué);2017年

9 馬蔚蓉;益腎填精法對(duì)排卵障礙性不孕癥卵泡發(fā)育潛能的臨床研究[D];南京中醫(yī)藥大學(xué);2012年

10 湯曉芳;小鼠卵巢Ski表達(dá)水平與卵泡發(fā)育的相關(guān)性研究[D];山東師范大學(xué);2011年

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