本文選題：GABA信號(hào) + 胚胎植入　； 參考：《重慶醫(yī)科大學(xué)》2011年碩士論文

【摘要】：目的胚胎植入和胎盤發(fā)生是胎生哺乳動(dòng)物及人類生殖的重要環(huán)節(jié)。胚胎植入的成功與否是決定胚胎能否進(jìn)一步發(fā)育的門檻,涉及母體全身的激素調(diào)控以及局部的胚胎與子宮間復(fù)雜的信息交流;植入后,胚胎滋養(yǎng)層細(xì)胞進(jìn)一步侵入子宮內(nèi)膜和血管,對胎盤形成及母胎血液循環(huán)至關(guān)重要,此過程具有嚴(yán)格的時(shí)空限制和生理自限性。比較研究發(fā)現(xiàn),胚胎植入過程、滋養(yǎng)層細(xì)胞的侵襲特性與腫瘤細(xì)胞的轉(zhuǎn)移和侵襲十分相似,其調(diào)節(jié)機(jī)制也近似。近年來發(fā)現(xiàn),γ-氨基丁酸(GABA)及其受體GABAAR、GABABR和合成酶在外周多種內(nèi)分泌組織和器官有特異性表達(dá),參與調(diào)節(jié)腫瘤細(xì)胞的侵襲和轉(zhuǎn)移。已有研究表明人和大鼠子宮組織表達(dá)GABA及其受體,特別是GABAA受體的π亞基在人著床窗口期子宮內(nèi)膜高表達(dá),提示其可能與子宮內(nèi)膜容受性改變和滋養(yǎng)層細(xì)胞侵襲有關(guān)。為探索GABA信號(hào)(GABA/GABAAR/GABABBR)在胚胎植入和胎盤發(fā)生中的作用,本研究采用免疫組織化學(xué)、胚胎體外培養(yǎng)、細(xì)胞侵襲實(shí)驗(yàn)等方法檢測GABA信號(hào)在小鼠早期胚胎植入和中期胎盤發(fā)生過程中的表達(dá)情況,并初步探討其在胚胎發(fā)育和滋養(yǎng)層細(xì)胞侵襲過程中的作用,為進(jìn)一步研究GABA信號(hào)與胚胎植入和胎盤發(fā)生提供線索。 方法1.免疫組織化學(xué)方法檢測GABA、GABRP、GB1蛋白在妊娠小鼠D1-D5子宮的表達(dá)情況;2.將小鼠囊胚培養(yǎng)于含不同濃度GABA抗體的培養(yǎng)液中,觀察囊胚脫帶、粘附和外延生長的情況;3.免疫組織化學(xué)方法檢測GABA、GABRP、GB1蛋白在妊娠小鼠D8-D12子宮和胚胎的表達(dá)及在絨毛膜滋養(yǎng)細(xì)胞癌來源的JAR細(xì)胞的表達(dá)情況;4.Matrigel細(xì)胞侵襲模型檢測GABA受體激動(dòng)劑和拮抗劑對JAR細(xì)胞侵襲遷移能力的影響。 結(jié)果1. GABA、GABRP、GB1蛋白在妊娠小鼠D1-D5子宮組織均有不同程度的表達(dá),且均于D4表達(dá)很強(qiáng),于D5特異性表達(dá)在胚胎植入位點(diǎn)的基質(zhì)細(xì)胞。2.一定濃度的GABA抗體能明顯抑制囊胚的脫帶率,提高已脫帶胚胎的外延生長面積,且呈劑量依賴效應(yīng),抗體濃度越大,作用越明顯;但對已脫帶囊胚的黏附率和外延生長率無明顯影響。3. GABA、GABRP、GB1蛋白在妊娠小鼠D8-D12的胚胎、胎盤錐、滋養(yǎng)層細(xì)胞以及子宮蛻膜組織和血管均有不同程度的表達(dá),并隨妊娠的進(jìn)程而變化。4. JAR細(xì)胞表達(dá)GABA、GABRP、GB1蛋白。5. GABAAR激動(dòng)劑muscimol和GABABR拮抗劑2-Hydroxysaclofen均顯著促進(jìn)JAR細(xì)胞對matrigel的侵襲;GABAAR拮抗劑SR-95531和GABABR激動(dòng)劑baclofen均顯著抑制JAR細(xì)胞對matrigel的侵襲。 結(jié)論1植入早期小鼠子宮表達(dá)GABA信號(hào),GABA抗體能影響囊胚的脫帶和外延生長,提示GABA信號(hào)可能參與植入過程中子宮與胚胎的“對話”,在小鼠胚胎著床中發(fā)揮作用。2妊娠中期小鼠子宮、胚胎以及母胎界面表達(dá)GABA信號(hào),激活GABAAR或阻斷GABABR能顯著提高滋養(yǎng)層細(xì)胞的侵襲能力,激活GABABR或阻斷GABAAR顯著降低滋養(yǎng)層細(xì)胞的侵襲能力,提示滋養(yǎng)層細(xì)胞可能通過自分泌、旁分泌和血液運(yùn)輸GABA,激活相應(yīng)的受體,協(xié)同調(diào)節(jié)滋養(yǎng)層細(xì)胞的侵襲,影響胎盤的發(fā)生。
[Abstract]:The purpose of embryo implantation and placentation is an important link of placental mammals and human reproduction. Embryo implantation success is decided whether to further the development of embryonic threshold, hormone regulation relates to maternal systemic and the embryo and the uterus local complex information exchange; implanted embryonic trophoblast cells, further invasion of the uterus and blood vessels of the formation of placenta and fetal blood circulation is crucial, this process has a strict time limit and physiological selflimited. Comparative study found that the process of embryo implantation, invasion and metastasis of tumor cells and invasion of trophoblast cells is very similar, the adjustment mechanism is similar. In recent years, gamma aminobutyric acid (GABA) and its receptor GABAAR, GABABR and synthetase specific expression in a variety of peripheral endocrine tissues and organs involved in the regulation of tumor cell invasion and metastasis. There have been studies The expression of GABA and its receptor in the rat uterus and the Ming Dynasty, especially the PI subunit of the GABAA receptor expression was high in endometrium, suggesting that it may be related to the invasion of endometrial receptivity and trophoblast cells. To explore the change of GABA signal (GABA/GABAAR/GABABBR) in the event of embryo implantation and placenta in the role of the immunohistochemistry, embryo culture, cell invasion assay method to detect the GABA signal in the early mouse embryo implantation and placental expression of medium in the process, and discuss the support layer of cells in the embryonic development and nourishing the role in the invasion process, to provide clues for further study of GABA signal and embryo implantation and placenta.
Methods: 1. immunohistochemical detection of GABA, GABRP, GB1 protein expression in D1-D5 mice uterine pregnancy; 2. cultured mouse blastocysts cultured in different concentration of GABA antibody, observe the blastocyst adhesion and removal, epitaxial growth of 3.; immunohistochemical detection of GABA, GABRP, GB1 protein expression in the expression of D8-D12 in mice uterus and embryo pregnancy and JAR cells in chorionic trophoblast cell carcinoma derived 4.Matrigel cells invasion model; effect of detection of GABA receptor agonists and antagonists on the invasion and migration of JAR cells.
Results 1. GABA, GABRP, GB1 protein expression in uterine tissues of pregnant D1-D5 mice at different levels, and on the expression of D4 is very strong, in the D5 specific expression of GABA antibody in the stromal cells of.2. certain concentration of embryo implantation sites can significantly inhibit the removal rate of blastocysts with, improve embryo growth area with the extension off and, in a dose-dependent manner, antibody concentration increases, the effect is more obvious; but to have off with the blastocyst adhesion rate and epitaxial growth rate has no obvious effect on.3. GABA, GABRP, GB1 protein in placenta cone of pregnant mouse D8-D12 embryos, trophoblast cells and decidual tissue and blood vessels have different levels of expression, and as the pregnancy process and the changes of.4. JAR cells expression of GABA, GABRP, GB1 protein.5. GABAAR agonist muscimol and GABABR antagonist 2-Hydroxysaclofen significantly promotes the invasion of JAR cells to Matrigel; GABAAR antagonists SR-95531 and GABABR The agonist baclofen significantly inhibited the invasion of JAR cells to Matrigel.
Conclusion implantation of mouse uterus 1 early expression of GABA signal, GABA antibody can affect the removal and epitaxial growth of blastocysts, suggesting that GABA signaling may be involved in the uterus and embryo implantation in the process of "dialogue", play the role of.2 in mouse uterine pregnancy mouse embryo implantation, embryo and maternal fetal interface expression of GABA signal, GABAAR activation or blockade of GABABR can significantly improve the trophoblast cell invasion ability, activation of GABABR or blocking GABAAR significantly reduced the invasive ability of trophoblast cells, suggesting that trophoblast cells via autocrine, paracrine and blood transport GABA, activate the corresponding receptor, CO regulation of trophoblast invasion, the effect of placenta.

【學(xué)位授予單位】：重慶醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2011
【分類號(hào)】：R321

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相關(guān)期刊論文 前2條

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本文編號(hào)：1770964