流感病毒致心脏发育异常的表观遗传分子机制研究
发布时间：2018-09-18 19:18
【摘要】：先天性心脏病(先心病)是一类严重危害人类健康的疾病,在新生儿中的发生率为4.05～12.3%。环境因素与遗传因素是先天性心脏病发生的主要原因。其中,孕早期宫内病毒感染是最为重要的环境因素之一。临床流行病学调查表明,有15种病毒的早期宫内感染可能会引起心脏发育异常,进而导致先心病的发生。近年来关于先天性心脏病的发生的遗传机制取得了许多进展,但是目前对病毒引起先天性心脏病的分子发生机制还不清楚。 本研究以BALB/c小鼠作为模式动物,在小鼠胚胎发育E7.5天通过鼻腔滴加的方法对孕母鼠进行H1N1流感病毒感染,构建H1N1流感病毒小鼠模型。通过数字基因表达谱技术筛选,发现在感染流感病毒后胚胎心脏的表观遗传相关基因表达出现差异。选取BAF染色质重塑复合物及其信号途径为研究对象,从RNA以及蛋白水平验证分析BAF染色质重塑相关基因及其下游Wnt信号和部分心脏发育关键基因的表达变化。 本文得到了以下研究结果： 1、利用数字化表达谱技术分析病毒组和正常组中表观遗传相关基因的表达情况,并进行筛选,经RT-PCR半定量分析发现25个表观遗传相关基因的结果与表达谱的结果一致,其中11个基因表达上调,14个基因表达下调。 2、表观遗传相关基因中BAF染色质重塑复合物出现显著性差异表达,并筛选出BAF染色质重塑基因及其下游Wnt信号和部分心脏发育关键基因进行进一步研究。 3、对BAF染色质重塑相关基因进行RT-PCR半定量分析,以E11.5天的胚胎心脏为材料,得到一致的RT-PCR结果：11组基因结果与表达谱的结果一致,并且都是下调的。 4、对Wnt信号下游基因进行蛋白表达水平分析,发现在病毒组中Sfrp2和Gata4的表达与数字表达谱的结果一致,均出现了下调。 本论文利用数字化表达谱技术筛选到在H1N1流感病毒感染孕鼠的胚胎心脏中表观遗传相关基因出现了显著差异表达。并且BAF染色质重塑复合物及其下游Wnt信号和部分心脏发育关键基因的表达也出现下调。提示表观遗传染色质重塑基因及其信号途径可能是流感病毒感染导致先天性心脏病发生的一个重要分子调控途径,本论文为进一步深入研究环境因素致先心病发生的分子机制研究提供了一定的前期理论基础。
[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 卤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.
【学位授予单位】：湖南师范大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R511.7;R541.1
本文编号：2248861
[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 卤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.
【学位授予单位】：湖南师范大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R511.7;R541.1
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