缺氧诱导因子-1α与冠状动脉缺血性心肌病的关联性研究

发布时间：2018-07-02 22:00

本文选题：缺血性心肌病 + 基因表达谱　；参考：《贵州医科大学》2017年硕士论文

【摘要】：目的:1、利用生物信息学工具对缺血性心肌病的基因芯片进行分析,为后续实验研究筛选出有意义的基因;2、探讨在排除混杂因素和效应修饰因子后,缺氧诱导因子-1α的表达对缺血性心肌病发病的独立作用大小,从而为缺血性心肌病的病因学研究和法医病理学鉴定提供新思路。方法:1.在美国国立生物信息中心(NCBI)公共基因芯片数据库(GEO)中下载缺血性心肌病的相关基因芯片数据,数据进行标准版处理后,采用Limma(R语言软件包)统计检验计算缺血性心肌病组ICM与正常样本组NF的显著性P值,采用BenjaminiHochberg方法进行多重检验纠正计算FDR,根据FDR值小于等于0.05来筛选差异基因。对差异化基因最近邻基因进行GO功能富集分析及KEGG Pathway功能富集分析,最后利用STRING构建蛋白质-蛋白质相互作用网络;2.收集贵州医科大学法医司法鉴定中心于2013-2015年尸检病例,纳入标准为:病理学诊断证实有不同程度冠状动脉狭窄,或生前已确诊为罹患冠心病者;排除标准:合并其他类型心肌病;死亡至尸检时间72小时。3.以是否发生缺血性心肌病作为分组,通过独立样本T检验、曼惠特U检验、卡方检验等统计学方法,对相关临床特征进行描述分析。4.对符合纳入标准的心肌组织进行HE染色,光镜观察心肌病理变化;免疫组化染色观察缺氧诱导因子-1α在心肌细胞核中的表达情况。5.建立多个回归方程模型,对缺氧诱导因子-1α与缺血性心肌病的独立效应值大小进行评估。结果:1.共找到差异基因1358个,参与细胞周期,缝隙链接、P53等生物过程及信号通路;将差异表达明显的基因构建的蛋白互作网络,发现HIF1α基因位于网络的中心;2.在控制混杂因素和效应修饰因子的条件下,HIF-1a对缺血性心肌病的独立作用为:以HIF-1a表达阴性组作为参照,弱阳性样本发生缺血性心肌病的概率病未提高(P均0.05)。而阳性、强阳性样本与阴性样本相比,ICM的发生率提高了3.7及7.5倍(模型1);而模型2则分别提高4.2及8.4倍。结论:1.应用生物信息学软件可在基因芯片的数据筛选出缺血性心肌病中的关键蛋白并进行分析。2.HIF-1a在参与缺血性心肌病发生发展过程的蛋白中起着关键连接的作用,它与其他周围基因产物之间关系都比较密切,进一步说明缺血性心肌病发生发展是多种蛋白相互作用的结果;3.HIF-1α与ICM的发生具有相关性。
[Abstract]:Objective to analyze the gene microarray of ischemic cardiomyopathy by bioinformatics, and to screen out a meaningful gene for further experimental study, and to explore the possibility of eliminating confounding factors and effect modifiers. The expression of hypoxia inducible factor-1 伪 (HIF-1 伪) plays an independent role in the pathogenesis of ischemic cardiomyopathy, which provides a new idea for etiological study and forensic pathological identification of ischemic cardiomyopathy. Method 1: 1. Download the gene-chip data related to ischemic cardiomyopathy in the National Bioinformatics Center (NCBI) Common GeneChip Database (GEO). The significant P value of NF in ICM group and normal sample group was calculated by Limma (R language package) statistical test, and FDR was calculated by Benjamin Hochberg method. The difference gene was screened according to FDR < 0.05. The differential gene nearest neighbor gene was analyzed by go function enrichment analysis and KEGG Pathway function enrichment analysis. Finally, protein-protein interaction network was constructed by STRING. The autopsy cases were collected from the Forensic Forensic Forensic Identification Center of Guizhou Medical University in 2013-2015. The included criteria were as follows: pathological diagnosis confirmed coronary artery stenosis in varying degrees or had been diagnosed as coronary heart disease before death; Exclusion criteria: other types of cardiomyopathy; death to autopsy time 72 hours. 3. With whether ischemic cardiomyopathy as a group, independent sample T test, Manette U test, chi-square test and other statistical methods to describe and analyze the clinical characteristics. 4. He staining was used to observe myocardial pathological changes under light microscope and the expression of hypoxia inducible factor-1 伪 in myocardial nuclei was observed by immunohistochemical staining. Several regression models were established to evaluate the independent effects of hypoxia inducible factor-1 伪 (HIF-1 伪) on ischemic cardiomyopathy. The result is 1: 1. A total of 1358 differentially expressed genes were found to be involved in biological processes and signaling pathways, such as cell cycle, gap link and p53, and the protein interaction network constructed by differentially expressed genes showed that HIF1 伪 gene was located at the center of the network. The independent effects of HIF-1a on ischemic cardiomyopathy under the control of confounding factors and effect modifiers were as follows: with HIF-1a negative expression group as control group, the probability of ischemic cardiomyopathy in weakly positive samples was not increased (P 0.05). The incidence of ICM in positive and strong positive samples was 3.7 and 7.5 times higher than that in negative samples (model 1), while that in model 2 was 4.2 and 8.4 times higher than that in negative samples. Conclusion 1. Using bioinformatics software, the key proteins in ischemic cardiomyopathy can be screened from the data of gene chip and analyzed. 2. HIF-1a plays a key role in the proteins involved in the pathogenesis and development of ischemic cardiomyopathy. The relationship between HIF-1 伪 and other peripheral gene products is very close, which further indicates that the occurrence and development of ischemic cardiomyopathy is the result of multiple protein interactions. 3. HIF-1 伪 is associated with ICM.
【学位授予单位】：贵州医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：D919

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