本文選題：骨形態(tài)發(fā)生蛋白-7(BMP-7) + 子宮內(nèi)膜��； 參考：《重慶醫(yī)科大學》2010年碩士論文

【摘要】： 目的:胚胎著床是胎生哺乳動物及人類生殖生理的重要環(huán)節(jié),是母體子宮與胚胎同步發(fā)育、在特定時期即“著床窗”開放期間精確調(diào)節(jié)、快速完成的生理現(xiàn)象。妊娠需要子宮組織發(fā)生廣泛的重建,子宮內(nèi)膜細胞生長和分化,即發(fā)生蛻膜化。子宮內(nèi)膜的蛻膜化過程對于成功的著床和妊娠的維持是必需的。蛻膜化過程涉及黏附分子、蛋白水解酶、生長因子、細胞因子、血管活性因子以及多種基因的相互作用和調(diào)節(jié)。已有研究表明BMPs家族在小鼠胚胎發(fā)育過程中都有表達,為進一步研究其家族中BMP-7在小鼠子宮中的表達規(guī)律和作用,本實驗采用細胞原代培養(yǎng)、熒光定量PCR、免疫組織化學、免疫細胞化學、Western blotting等方法從mRNA和蛋白水平檢測BMP-7在妊娠小鼠子宮中的表達,探索其在子宮內(nèi)膜蛻膜化過程中的作用。 方法: 1.RT-PCR和免疫組織化學技術分別觀察BMP-7的mRNA和蛋白在妊娠第5-8日小鼠子宮中的表達情況;2.原代培養(yǎng)小鼠子宮基質(zhì)細胞,用孕酮、雌二醇和cAMP誘導蛻膜化,并在誘導蛻膜化的同時分別加入1.0ng/ml、10ng/ml、100ng/ml BMP-7重組腺病毒,于24h、48h、72h、96h檢測催乳素的表達;3.去卵巢小鼠隨機分3組,分別皮下注射芝麻油(0.1ml/只)、孕酮(2.0mg/只)、RU486(2.0mg/只)+孕酮(2.0mg/只),于注射后0h、6h、12h、24h處死小鼠收集子宮,提取子宮RNA和總蛋白,熒光定量PCR、Western blotting檢測BMP-7的表達情況。 結果:1.妊娠第5-8日,BMP-7 mRNA在小鼠子宮著床位點的表達高于著床旁組織(P0.01),且在著床點的表達隨著妊娠天數(shù)的增加逐漸增強(P0.05)。妊娠第5日、第6日,BMP-7蛋白分別表達于植入胚胎周圍的基質(zhì)細胞和初級蛻膜帶;第7日、第8日,主要表達于次級蛻膜帶以及植入位點系膜側基質(zhì)細胞。2.在小鼠子宮基質(zhì)細胞的蛻膜化過程中,BMP-7重組腺病毒組催乳素的mRNA和蛋白表達均顯著增加(P0.01),其中10ng/ml組增加最明顯(P0.01)。3.去卵巢小鼠模型研究顯示:孕酮上調(diào)了小鼠子宮組織BMP-7的表達(P0.01),孕酮受體拮抗劑RU486能有效阻斷孕酮對BMP-7的上調(diào)作用(P0.05) 結論:BMP-7基因在小鼠子宮內(nèi)的表達受孕酮調(diào)控,具有促進小鼠子宮內(nèi)膜蛻膜化的效應。
[Abstract]:Objective: embryo implantation is an important link in reproductive physiology of fetal mammals and human beings. It is a physiological phenomenon that the womb and embryo develop synchronously and regulate accurately and finish quickly during the opening period of implantation window. Pregnancy requires extensive reconstruction of uterine tissue, endometrial cell growth and differentiation, that is, decidualization. Decidualization of the endometrium is essential for successful implantation and pregnancy maintenance. Decidualization involves the interaction and regulation of adhesion molecules, proteolytic enzymes, growth factors, cytokines, vasoactive factors and multiple genes. In order to further study the expression of BMP-7 in mouse uterus, the primary cell culture, fluorescence quantitative PCR and immunohistochemistry were used to study the expression of BMP-7 in mouse uterus. The expression of BMP-7 in the uterus of pregnant mice was detected by immunocytochemistry and Western blotting, and the role of BMP-7 in decidualization of endometrium was explored. Methods: 1. RT-PCR and immunohistochemistry were used to observe the expression of BMP-7 mRNA and protein in the uterus of mice on the 5th and 8th day of pregnancy respectively. Primary cultured mouse uterine stromal cells were induced decidualization by progesterone, estradiol and camp. The recombinant adenovirus BMP-7 was added with 1.0 ng / ml 10ng / ml BMP-7 respectively. Prolactin expression was detected at 24 h, 48 h, 72 h and 96 h, respectively. Ovariectomized mice were randomly divided into three groups: subcutaneously injected with sesame oil (0.1ml/), progesterone (2.0mg/), RU486 (2.0mg/), progesterone (2.0mg/). The mice were killed at 0 h, 6 h, 12 h and 24 h after injection to collect uterus, extract uterine RNA and total protein, and detect the expression of BMP-7 by Western blotting. The result is 1: 1. The expression of BMP-7 mRNA on the 5th to 8th day of pregnancy was higher than that in the para-implantation tissue (P0.01), and the expression of BMP-7 mRNA increased gradually with the increase of pregnancy days (P0.05). On the 5th and 6th day of pregnancy, BMP-7 protein was expressed in stromal cells and primary decidua band around implantation embryos, and mainly in secondary decidua band and Mesangial side stromal cells at implantation site on the 7th and 8th day of gestation. During decidualization of mouse uterine stromal cells, prolactin mRNA and protein expression in BMP-7 recombinant adenovirus group increased significantly (P0.01), especially in 10ng/ml group (P0.01). The study of ovariectomized mouse model showed that progesterone up-regulated the expression of BMP-7 in mouse uterus (P0.01). RU486, a progesterone receptor antagonist, could effectively block the up-regulation of BMP-7 by progesterone (P0.05). The expression of progesterone is regulated by progesterone. It has the effect of promoting decidualization of mouse endometrium.
【學位授予單位】：重慶醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2010
【分類號】：R321

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本文編號：2058518