本文選題：子宮內(nèi)膜容受性 + 甲基化　； 參考：《重慶醫(yī)科大學(xué)》2012年博士論文

【摘要】：目的：葉酸是哺乳類動(dòng)物生長(zhǎng)發(fā)育過(guò)程中擔(dān)負(fù)一碳單位傳遞的B族維生素，，是機(jī)體各種甲基化反應(yīng)的甲基供體。葉酸攝入不足或葉酸代謝障礙時(shí)，可引起基因組DNA固有甲基化模式的改變而促進(jìn)基因表達(dá)或抑制基因表達(dá)，誘發(fā)疾病狀態(tài)，通過(guò)表觀遺傳修飾這一途徑參與多種生命活動(dòng)過(guò)程中的基因表達(dá)調(diào)控。但迄今為止，對(duì)于在妊娠早期子宮內(nèi)膜容受性建立過(guò)程中，低葉酸水平是否會(huì)通過(guò)表觀遺傳修飾（DNA甲基化）來(lái)調(diào)控子宮內(nèi)膜容受性相關(guān)基因的表達(dá)，進(jìn)而影響胚胎著床，目前未見任何相關(guān)報(bào)道。因此，本研究旨在探索孕期低葉酸水平是否會(huì)影響子宮內(nèi)膜容受性相關(guān)基因（Cdh1、Pgr、Esr1）DNA甲基化模式及其基因表達(dá)的改變，從而對(duì)胚胎著床產(chǎn)生影響。 方法：建立葉酸缺乏孕鼠和正常孕鼠胚胎著床模型。采用甲基化特異性PCR和亞硫酸氫鈉測(cè)序方法對(duì)Cdh1、Pgr、Esr1基因啟動(dòng)子區(qū)的甲基化模式進(jìn)行分析。采用Real-time PCR、Western-blot、免疫組化對(duì)Cdh1、Pgr、Esr1基因的mRNA和蛋白表達(dá)進(jìn)行分析。采用掃描電鏡方法對(duì)子宮內(nèi)膜容受性建立的形態(tài)學(xué)標(biāo)志物—胞飲突的形態(tài)和數(shù)量進(jìn)行檢測(cè)。孕鼠尾靜脈注射臺(tái)盼藍(lán)后計(jì)數(shù)孕鼠妊娠第5天的胚胎著床點(diǎn)數(shù)量。 結(jié)果:甲基化分析結(jié)果顯示葉酸缺乏情況下Esr1基因啟動(dòng)子區(qū)的甲基化率顯著降低，而Cdh1、Pgr基因啟動(dòng)子的甲基化率沒(méi)有發(fā)生顯著改變。基因表達(dá)分析結(jié)果顯示葉酸缺乏孕鼠組Esr1基因表達(dá)顯著降低，而Cdh1、Pgr基因表達(dá)略高于正常對(duì)照組，但不具有統(tǒng)計(jì)學(xué)意義。掃描電鏡結(jié)果顯示葉酸缺乏情況下胞飲突的數(shù)量和形態(tài)均未發(fā)生明顯改變。與正常對(duì)照組相比較，葉酸缺乏孕鼠組的妊娠率和妊娠第5天孕鼠的胚胎著床點(diǎn)數(shù)量均沒(méi)有發(fā)生顯著改變，胚胎正常著床。 結(jié)論：在子宮內(nèi)膜容受性建立這一正常生理過(guò)程中，DNA甲基化對(duì)容受性相關(guān)基因Cdh1、Pgr的表達(dá)調(diào)控作用并不明顯，而Esr1的表達(dá)可能會(huì)受到DNA甲基化調(diào)控，但并不影響胚胎正常著床。葉酸缺乏下胚胎能夠正常著床提示葉酸缺乏導(dǎo)致胚胎發(fā)育異�？赡苁侵惨院蟮呐咛グl(fā)育發(fā)生了異常。甲基化調(diào)控在腫瘤發(fā)生發(fā)展過(guò)程中與子宮內(nèi)膜容受性建立這一正常生理過(guò)程中的不同作用，可能會(huì)為闡明腫瘤侵襲的“無(wú)控性”和胚胎著床的“有控性”機(jī)制提供新的線索。 目的：胚胎著床以后子宮內(nèi)膜蛻膜功能的正常表達(dá)是胎盤形成的重要基礎(chǔ)，并在胎盤形成以前擔(dān)負(fù)著保護(hù)和營(yíng)養(yǎng)胚胎的重要作用，對(duì)妊娠的建立和維持至關(guān)重要。研究發(fā)現(xiàn)葉酸對(duì)胎盤的發(fā)育起著重要作用。但葉酸對(duì)著床后子宮內(nèi)膜蛻膜化進(jìn)程的影響以及葉酸缺乏是否會(huì)引起子宮內(nèi)膜蛻膜化發(fā)生改變，從而造成胚胎發(fā)育異常，目前并不清楚，這值得我們?nèi)ヌ剿�。因此，本研究旨在探索著床后子宮內(nèi)膜蛻膜化是否會(huì)受到孕期低葉酸水平的影響。 方法：建立葉酸缺乏孕鼠和正常孕鼠胚胎著床模型，對(duì)孕鼠胎兒的出生情況進(jìn)行統(tǒng)計(jì)。采用HE方法檢測(cè)孕鼠妊娠d7子宮內(nèi)膜形態(tài)。采用Real-time PCR、Western-blot、免疫組化對(duì)孕鼠妊娠d7、d8子宮內(nèi)膜Hoxa10、MMP2基因表達(dá)進(jìn)行分析。采用流式細(xì)胞術(shù)檢測(cè)妊娠d7子宮內(nèi)膜的凋亡情況，并采用Western-blot對(duì)凋亡相關(guān)因子Bax、Bcl2的表達(dá)進(jìn)行了分析。 結(jié)果:葉酸缺乏孕鼠組胎兒出生率為（0，0/20）顯著低于正常對(duì)照組（90%，18/20）。觀察妊娠d7孕鼠子宮，葉酸缺乏孕鼠組（50%，4/8）胚胎呈現(xiàn)不同程度的吸收和發(fā)育阻滯現(xiàn)象，明顯高于正常對(duì)照組（12.5%，1/8）。HE染色觀察發(fā)現(xiàn)妊娠d7葉酸缺乏孕鼠組子宮內(nèi)膜蛻膜形態(tài)發(fā)生改變。進(jìn)一步檢測(cè)發(fā)現(xiàn)葉酸缺乏情況下，在蛻膜化過(guò)程中起著重要作用的Hoxa10和MMP2基因表達(dá)顯著降低。凋亡的正常維持對(duì)蛻膜化具有重要意義。檢測(cè)發(fā)現(xiàn)葉酸缺乏孕鼠組妊娠d7子宮內(nèi)膜的凋亡率明顯低于正常對(duì)照組，且葉酸缺乏孕鼠組促凋亡因子Bax蛋白表達(dá)明顯低于正常組，而抑凋亡因子Bcl2的表達(dá)卻顯著高于正常組。 結(jié)論：研究發(fā)現(xiàn)葉酸缺乏情況下，在子宮內(nèi)膜的蛻膜化中起著重要作用的Hoxa10、MMP2基因以及維持蛻膜化凋亡平衡的相關(guān)基因Bax、Bcl2表達(dá)均發(fā)生改變，子宮內(nèi)膜蛻膜化受到損害，提示葉酸在子宮內(nèi)膜蛻膜化過(guò)程中發(fā)揮著重要作用。
[Abstract]:Objective : To investigate whether folic acid is a group B vitamin which is transmitted by a carbon unit during the growth and development of mammals , which is a methyl donor with various methylation reactions .

Methods : The methylation patterns of Cdh1 , Pgr , Esr1 gene were analyzed by means of methylation specific PCR and sodium bisulfite sequencing . The expression of mRNA and protein of Cdh1 , Pgr , Esr1 gene was analyzed by means of real - time PCR , Western - blot and immunohistochemistry . The morphology and quantity of the cell - drinking process were detected by scanning electron microscopy .

Results : methylation analysis showed that the methylation rate of Esr1 gene promoter region decreased significantly in the absence of folic acid , but the methylation rate of Cdh1 and Pgr gene promoter did not change significantly . The expression of Cdh1 and Pgr gene was slightly higher than that of the normal control group , but the expression of Cdh1 and Pgr gene was slightly higher than that in the normal control group .

Conclusion : In this normal physiological process , the expression of DNA methylation may be regulated by DNA methylation , but the expression of Esr1 may be regulated by DNA methylation , but it does not affect the normal implantation of embryo .

Objective : To investigate whether folic acid plays an important role in the establishment and maintenance of placenta after implantation of embryo . It is very important for the establishment and maintenance of the placenta before the formation of placenta .

Methods : The embryo implantation model of pregnant rats and normal pregnant rats was established , and the birth of pregnant rats was analyzed by HE method . The expression of Hox10 and MMP2 in pregnant rats was analyzed by using HE method . The expression of apoptosis related factors Bax and Bcl 2 was analyzed by flow cytometry .

Results : The fetal birth rate of folic acid ( 0 , 0 / 20 ) was significantly lower than that of the control group ( 90 % , 18 / 20 ) .

Conclusion : In the absence of folic acid , the expression of Hox10 , MMP2 gene and the related genes Bax and Bcl 2 in decidualization of endometrium have been changed , and the expression of Bcl 2 and Bcl 2 in the endometrium have been altered . It is suggested that folic acid plays an important role in the process of decidualization of endometrium .

【學(xué)位授予單位】：重慶醫(yī)科大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2012
【分類號(hào)】：R321

【參考文獻(xiàn)】

相關(guān)期刊論文 前2條

1 倪錦;江艷芳;古妙寧;;葉酸和同型半胱氨酸在神經(jīng)系統(tǒng)發(fā)育中的作用研究進(jìn)展[J];中國(guó)兒童保健雜志;2010年11期

2 楊慧敏,廖國(guó)義,周冬,余靈,黃云祖,周天鴻;葉酸缺乏孕鼠子代腦組織DNA甲基化水平的變化[J];中國(guó)小兒血液;2003年02期

本文編號(hào)：1855907