五指山小型猪心肌梗塞相关非编码RNA的鉴定及其调控机制研究

Myocardial infarction is a common acute cardiovascular diseases and seriously harmful to human health. Non-coding RNAs(ncRNAs) which have become a hot topic of researches have very important regulatory function on the disease and recovery. Meanwhile minipig is one of the ideal laboratory animals for human cardiovascular diseases researches. The minipig myocardial infarction model is very important for the researches on the pathogenesis and treatment. On basis of the former researches, the new project will complete several new researches. At first we plan to make a high success ratio Wuzhishan Minipigs (WZSP) myocardial infarction model by surgical operation. The model will performe the proceeding and characteristics of the disease. After the animal model accomplished, we will screen and identify systematically the relationships between myocardial infarction disease & recovery and relative ncRNAs by means of transcriptome sequencing, mass spectrometry-based differential proteomics, bioinformatic analysis, Overexpression and inhibition of expression. We will also study the regulatory function of target genes on the occurrence and prognosis of the disease. All the researches will try to establish a foundation to illustrate the molecular regulatory mechanism about myocardial infarction disease on minipig model, futher more to provide new theories for pathogenesis and prevention and treatment of human myocardial infarction disease.

心肌梗塞是常见的急性心血管内科疾病,严重危害人类的健康。而非编码RNA在心肌梗塞与重塑中具有十分重要的调控功能，已成当今研究热点之一。其中，小型猪动物模型对掌握人类心梗发病机理与治疗研究具有重要意义，是理想的人类心血管研究实验动物。本项目在前期工作的基础上，通过手术方法模拟人类心梗发病特点，建立起一个能体现疾病发展进程和特点的、成功率高的心梗动物模型；并利用转录组和差异蛋白质组测序、生物信息学分析、过表达和表达抑制等技术与手段，在已建成的动物模型的基础上，通过对心肌梗塞相关非编码RNA（miRNAs）的系统筛选与鉴定，及其与靶基因在疾病发生与转归过程中的调控功能的研究，为阐明小型猪心肌梗塞的分子调控机理奠定基础，也为进一步研究人类心梗发病机制及其预防治疗提供新依据。

心肌梗塞是常见的急性心血管内科疾病，常危及生命。研究表明，miRNAs在心肌梗塞中具有十分重要的调控功能，相关基因和miRNAs影响心脏功能，进而影响心肌梗塞后血管再生与纤维化进程。.项目在利用我国特有小型猪资源手术结扎冠状动脉法成功建立小型猪心肌梗塞模型的基础上，通过转录组测序，结合生物信息学方法，对五指山小型猪心肌梗塞相关miRNAs及靶基因进行筛查与鉴定，筛选出7个重要的miRNAs及部分靶基因。利用人脐静脉内皮细胞（HUVEC）模型和人心脏成纤维细胞（HCF）模型，采用Edu荧光显微镜检测、Transwell assay、qRT-PCR、Western blot、双荧光素酶报告基因验证等分子生物学手段，研究了筛选出的7个重要miRNAs对心肌梗塞后血管再生与心肌纤维化的作用。.miR-21-3p抑制HUVEC增殖，miR-27b-3p促进HUVEC增殖，miR-144-3p、miR-21-3p、miR-142-5p、miR-27b-3p均抑制HUVEC迁移及血管腔形成。4个miRNAs通过影响HUVEC的增殖、迁移及血管腔形成参与心肌梗塞后血管再生过程。.miR-144-3p靶向作用于PTEN，诱导HCF增殖、迁移并转化为肌成纤维细胞，促进心肌Ⅰ、Ⅲ型胶原蓄积；miR-590-3p靶向作用于ZEB1，抑制HCF增殖、迁移和向肌成纤维细胞转化，抑制心肌Ⅰ、Ⅲ型胶原蓄积。2个miRNAs通过影响HCF的增殖、迁移、转化及胶原合成参与心肌梗塞后心肌纤维化过程。.项目从分子水平及细胞水平探究了这些miRNAs的生物学功能及其与靶基因的相互作用关系，为探索心肌梗塞的分子调控机制提供了miRNA层面的理论基础，进而为阐明心肌梗塞的分子调控机制打下基础，也为进一步开展相关研究提供了依据。