【摘要】：先天性心臟病(先心病)是一類嚴(yán)重危害人類健康的疾病,在新生兒中的發(fā)生率為4.05～12.3%。環(huán)境因素與遺傳因素是先天性心臟病發(fā)生的主要原因。其中,孕早期宮內(nèi)病毒感染是最為重要的環(huán)境因素之一。臨床流行病學(xué)調(diào)查表明,有15種病毒的早期宮內(nèi)感染可能會(huì)引起心臟發(fā)育異常,進(jìn)而導(dǎo)致先心病的發(fā)生。近年來(lái)關(guān)于先天性心臟病的發(fā)生的遺傳機(jī)制取得了許多進(jìn)展,但是目前對(duì)病毒引起先天性心臟病的分子發(fā)生機(jī)制還不清楚。 本研究以BALB/c小鼠作為模式動(dòng)物,在小鼠胚胎發(fā)育E7.5天通過(guò)鼻腔滴加的方法對(duì)孕母鼠進(jìn)行H1N1流感病毒感染,構(gòu)建H1N1流感病毒小鼠模型。通過(guò)數(shù)字基因表達(dá)譜技術(shù)篩選,發(fā)現(xiàn)在感染流感病毒后胚胎心臟的表觀遺傳相關(guān)基因表達(dá)出現(xiàn)差異。選取BAF染色質(zhì)重塑復(fù)合物及其信號(hào)途徑為研究對(duì)象,從RNA以及蛋白水平驗(yàn)證分析BAF染色質(zhì)重塑相關(guān)基因及其下游Wnt信號(hào)和部分心臟發(fā)育關(guān)鍵基因的表達(dá)變化。 本文得到了以下研究結(jié)果： 1、利用數(shù)字化表達(dá)譜技術(shù)分析病毒組和正常組中表觀遺傳相關(guān)基因的表達(dá)情況,并進(jìn)行篩選,經(jīng)RT-PCR半定量分析發(fā)現(xiàn)25個(gè)表觀遺傳相關(guān)基因的結(jié)果與表達(dá)譜的結(jié)果一致,其中11個(gè)基因表達(dá)上調(diào),14個(gè)基因表達(dá)下調(diào)。 2、表觀遺傳相關(guān)基因中BAF染色質(zhì)重塑復(fù)合物出現(xiàn)顯著性差異表達(dá),并篩選出BAF染色質(zhì)重塑基因及其下游Wnt信號(hào)和部分心臟發(fā)育關(guān)鍵基因進(jìn)行進(jìn)一步研究。 3、對(duì)BAF染色質(zhì)重塑相關(guān)基因進(jìn)行RT-PCR半定量分析,以E11.5天的胚胎心臟為材料,得到一致的RT-PCR結(jié)果：11組基因結(jié)果與表達(dá)譜的結(jié)果一致,并且都是下調(diào)的。 4、對(duì)Wnt信號(hào)下游基因進(jìn)行蛋白表達(dá)水平分析,發(fā)現(xiàn)在病毒組中Sfrp2和Gata4的表達(dá)與數(shù)字表達(dá)譜的結(jié)果一致,均出現(xiàn)了下調(diào)。 本論文利用數(shù)字化表達(dá)譜技術(shù)篩選到在H1N1流感病毒感染孕鼠的胚胎心臟中表觀遺傳相關(guān)基因出現(xiàn)了顯著差異表達(dá)。并且BAF染色質(zhì)重塑復(fù)合物及其下游Wnt信號(hào)和部分心臟發(fā)育關(guān)鍵基因的表達(dá)也出現(xiàn)下調(diào)。提示表觀遺傳染色質(zhì)重塑基因及其信號(hào)途徑可能是流感病毒感染導(dǎo)致先天性心臟病發(fā)生的一個(gè)重要分子調(diào)控途徑,本論文為進(jìn)一步深入研究環(huán)境因素致先心病發(fā)生的分子機(jī)制研究提供了一定的前期理論基礎(chǔ)。
[Abstract]:Congenital heart disease (CHD) is a kind of serious disease which is harmful to human health. The incidence of congenital heart disease in newborn is 4.05 鹵12.3. Environmental and genetic factors are the main causes of congenital heart disease. Intrauterine virus infection in early pregnancy is one of the most important environmental factors. Clinical epidemiological investigation showed that early intrauterine infection of 15 viruses may lead to cardiac dysplasia, which may lead to congenital heart disease. In recent years, many advances have been made in the genetic mechanism of congenital heart disease, but the molecular mechanism of virus induced congenital heart disease is still unclear. In this study, BALB/c mice were used as model animals. The pregnant female mice were infected with H1N1 influenza virus by nasal drip on day 7.5 of embryonic development of mice. The mouse model of H1N1 influenza virus was established. The expression of epigenetic genes in the embryonic heart after influenza virus infection was found to be different by digital gene expression profiling. BAF chromatin remodeling complex and its signaling pathway were selected to analyze the expression changes of BAF chromatin remodeling related genes, their downstream Wnt signals and some key cardiac development genes from RNA and protein levels. The results are as follows: 1. The expression of epigenetic genes in virus group and normal group was analyzed and screened by digital expression profiling. By semi-quantitative analysis of RT-PCR, the results of 25 epigenetic related genes were consistent with those of expression profile. Among them, 11 genes were up-regulated and 14 genes were down-regulated. 2. There was significant difference in expression of BAF chromatin remodeling complex in epigenetic related genes. The BAF chromatin remodeling gene, its downstream Wnt signal and some key genes of cardiac development were screened for further study. 3. The BAF chromatin remodeling related genes were semi-quantitatively analyzed by RT-PCR. The embryonic heart of E11.5 day was used as the material. The results of RT-PCR: 11 groups of genes were consistent with the results of the expression profile, and they were all down-regulated. The protein expression level of downstream genes of Wnt signal was analyzed. It was found that the expression of Sfrp2 and Gata4 in the virus group was consistent with the results of the digital expression profile, and both of them were down-regulated. In this study, the differential expression of epigenetic genes in fetal heart of pregnant mice infected with H1N1 influenza virus was screened by digital expression profiling. The expression of BAF chromatin remodeling complex and its downstream Wnt signal and some heart development key genes were also down-regulated. The results suggest that epigenetic chromatin remodeling gene and its signaling pathway may be an important molecular regulatory pathway of congenital heart disease caused by influenza virus infection. This paper provides a theoretical basis for the further study of the molecular mechanism of environmental factors causing congenital heart disease.
【學(xué)位授予單位】：湖南師范大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類號(hào)】：R511.7;R541.1

【參考文獻(xiàn)】

相關(guān)期刊論文 前5條

1 崔君兆;我國(guó)孕婦弓形體感染致胎嬰先天畸形缺陷的監(jiān)測(cè)[J];疾病監(jiān)測(cè);1996年08期

2 林兆宇;高翔;;小鼠的遺傳學(xué)研究[J];生命科學(xué);2006年05期

3 王凱;;生命科學(xué)研究中常用模式生物[J];生命科學(xué)研究;2010年02期

4 朱軍;出生缺陷及其監(jiān)測(cè)[J];中國(guó)實(shí)用婦科與產(chǎn)科雜志;2002年09期

5 殷勝利,張希,孫培吾,王治平,羅紅鶴;先天性心臟病的染色體研究[J];中華實(shí)驗(yàn)外科雜志;2001年06期

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