小鼠酒精性心肌病相关非编码RNAs表达谱特征分析

发布时间：2018-05-06 10:12

本文选题：酒精 + 心肌病　；参考：《昆明医科大学》2015年硕士论文

【摘要】：目的：长期过量饮酒可引起机体多个重要器官发生细胞学、组织学及其结构的改变,导致胃溃疡,胰腺炎,酒精性脂肪肝,酒精性心肌病等疾病的发生。本论文以酒精性心肌病为研究对象,利用高通量基因芯片检测长期过量饮酒小鼠心肌组织中差异表达的microRNAs, long non-coding RNAs (lnc RNAs)和circular RNAs等非编码RNAs的表达谱,从分子生物学角度探索酒精性心肌病的发生机制。方法：100只昆明种小鼠(12～14周龄),随机分为饮酒组(n=60)和对照组(n=40),4%的乙醇水溶液作为饮酒组小鼠唯一的饮用水源,采用自由饮用的方式饲喂24周制作酒精性心肌病模型。持续建模24周后,随机摘取饮酒组和对照组小鼠心脏各5颗制作石蜡切片和超薄切片,分别在光学显微镜和透射电子显微镜下观察心脏的病理学改变和心肌细胞超微结构改变；确认小鼠酒精性心肌病模型建成后,利用基因芯片检测实验组和对照组各3例心脏样本中的microRNAs, lncRNAs ,circular RNAs和mRNAs表达谱。通过对原始数据进行均一化预处理后,确定酒精性心肌病心肌组织中具有明显差异表达的microRNAs, lnc RNAs, circular RNAs和mRNAs o然后利用生物信息学软件分析差异表达的mRNAs的基因功能和其参与的生物学信号通路,结合文献找出与酒精性心肌病发病机制关系最为密切的蛋白质编码基因mRNAs及其参与的信号通路。结果：1.光学显微镜下,饮酒组和对照组小鼠相同心脏横切面HE染色石蜡切片图像显示：饮酒组小鼠心脏室间隔明显增厚；透射电子显微镜下观察到饮酒组小鼠心肌细胞内肌原纤维溶解,肌节连续性中断,线粒体增多变形、内嵴退化,内质网肿胀等表现。2.通过对基因芯片原始数据进行预处理达到均一化后,与对照组相比,在饮酒组小鼠心脏组织中有19个microRNAs,494个lncRNAs,265个circular RNAs和404个mRNAs出现明显差异性表达(p0.05)。3.利用生物信息学分析工具对差异表达的mRNAs的基因功能及其参与的信号通路预测分析发现,在饮酒组小鼠心脏组织中差异表达的mRNAs主要参与碳水化合物代谢过程。结论：1.长期过量饮酒可以导致小鼠心脏重构,心肌细胞内肌原纤维,线粒体,内质网等超微结构改变；2.长期过量饮酒能引起小鼠心脏组织内大量非编码RNAs和mRNAs表达失调。
[Abstract]:Objective: excessive drinking for a long time can cause many important organogenesis cytology, histology and structural changes, leading to gastric ulcer, pancreatitis, alcoholic fatty liver, alcoholic cardiomyopathy and other diseases. In this study, alcoholic cardiomyopathy was used to detect the differential expression profiles of microRNas, long non-coding RNAs lnc RNAsand circular RNAs noncoding RNAs in myocardial tissues of long-drinking mice with high throughput gene microarray. To explore the mechanism of alcoholic cardiomyopathy from the point of molecular biology. Methods A total of 100 Kunming mice were randomly divided into drinking group (n = 60) and control group (n = 40%) as the only drinking water source. The model of alcoholic cardiomyopathy was established by free drinking for 24 weeks. After 24 weeks of continuous modeling, 5 hearts of drinking group and control group were randomly selected to make paraffin sections and ultrathin sections. The pathological changes and ultrastructure of cardiac myocytes were observed under optical microscope and transmission electron microscope respectively. After the establishment of mice model of alcoholic cardiomyopathy, microRNA RNAs, lncRNAs circular RNAs and mRNAs were detected by gene microarray in 3 heart samples from experimental group and 3 control group. After homogenizing the raw data, To identify microRNAs, lnc RNAs, circular RNAs and mRNAs o, which have obvious differential expression in myocardial tissue of alcoholic cardiomyopathy, and then to analyze the gene function and biological signal pathway of differentially expressed mRNAs using bioinformatics software. In combination with the literature, the protein encoding gene mRNAs, which is most closely related to the pathogenesis of alcoholic cardiomyopathy, and the signal pathway involved in it are found. The result is 1: 1. Under optical microscope, the paraffin sections of the same cross section of heart in drinking group and control group showed that the ventricular septum of drinking group was thicker than that of control group. Transmission electron microscope showed that the myofibrils dissolved, the continuity of sarcomere interrupted, the mitochondria increased and deformed, the inner ridge degenerated and the endoplasmic reticulum swelled in drinking group. Compared with the control group, there were 19 microRNASs 494 lncRNAss, 265 circular RNAs and 404 mRNAs in the drinking group, and the significant difference was found between the drinking group and the control group after homogenization of the original gene chip data. Using bioinformatics tools to predict the gene function of differentially expressed mRNAs and its involvement in signal pathways, it was found that the differentially expressed mRNAs in the heart of drinking mice was mainly involved in carbohydrate metabolism. Conclusion 1. Long-term excessive drinking could result in cardiac remodeling, ultrastructural changes of myofibrils, mitochondria and endoplasmic reticulum in cardiac myocytes. Excessive drinking for a long time caused a large number of non-coding RNAs and mRNAs expression disorders in the heart of mice.
【学位授予单位】：昆明医科大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R542.2

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