咳嗽变异性哮喘与典型支气管哮喘患者外周血单个核细胞全基因组表达谱的差异研究

发布时间：2018-06-13 02:36

  本文选题：咳嗽变异性哮喘 + 支气管哮喘　； 参考：《川北医学院》2017年硕士论文

【摘要】：目的:咳嗽变异性哮喘(Cough variant asthma,CVA)是一种不典型的支气管哮喘(Bronchial asthma,以下简称哮喘),因其可发展为典型哮喘,也被称为哮喘的前驱阶段,早期正确治疗可部分避免由CVA向典型哮喘的进一步发展。而CVA发展成为哮喘或正常的机制目前尚缺乏相关报道。基于此,本研究对CVA、典型哮喘以及正常人外周血单个核细胞(Peripheral Blood Mononuclear Cells,PBMCs)的差异表达基因进行分析。以期找出CVA异于典型哮喘及正常人PBMCs的基因表达谱,借此进一步阐述哮喘的发病机制及CVA的可能转归机制。方法:收集正常人、CVA患者及典型哮喘患者各5例,每组各例分别采集外周静脉血3ml,分离出PBMCs,并利用Trizol法提取总RNA,再用Agilent 4×44K人类全基因组Oligo芯片进行检测,并对其差异基因行聚类分析及Gene Ontology功能分析,随之挑选10个差异表达显著且功能相关的基因作为候选基因,然后利用实时荧光定量PCR(Real-Time PCR,RT-PCR)技术对候选基因进行验证。结果:基因芯片检测结果发现,哮喘组与CVA组比较,有差异表达基因202个,正常组与CVA组比较,有差异表达基因119个,正常组与哮喘组比较,有差异表达基因779个,差异均在2倍以上。Gene Ontology功能分析提示差异基因主要与表观遗传学及抗原处理和表达等相关。筛选10个候选基因,分别是HDC、EGR1、DEFA4、LTF、G0S2、IL4、TFF3、FCER1A、CAMP、CTSG。Real-Time PCR验证结果与基因芯片结果不完全一致。FCER1A、HDC、IL4基因在CVA组中较正常组以及哮喘组均表达上调(P0.05),其相对表达量分别为:(FCER1A)1.45×10-1±5.26×10-2、7.75×10-2±2.47×10-2、5.29×10-2±2.66×10-2;(HDC)2.86×10-2±1.47×10-2、8.89×10-3±3.4×10-3、7.44×10-3±5.29×10-3;(IL4)1.66×10-3±8.63×10-4、3.75×10-4±3.28×10-4、3.44×10-4±2.21×10-4。EGR1、DEFA4、LTF、G0S2、TFF3、CAMP、CTSG基因的表达在三组间比较差异均无统计学意义(P0.05)。结论:1、本研究人全基因组表达芯片结果显示CVA与典型哮喘及正常人PBMC的基因表达谱均存在明显差异。2、Real-Time PCR验证结果显示,FCER1A、HDC、IL4基因在CVA组中较正常组以及哮喘组均表达上调,EGR1、DEFA4、LTF、G0S2、TFF3、CAMP、CTSG基因在三组间的比较差异均无统计学意义。提示CVA的发病可能与FCER1A、HDC、IL4基因有关,而其余7个基因可能不是影响CVA发病及转归的重要基因。
[Abstract]:Objective: cough variant asthma (CVA) is an atypical bronchial asthma with Bronchial asthma (hereinafter referred to as Asthma), which can develop into typical asthma and is also known as the precursor stage of asthma. Early correct treatment may partly avoid further development from CVA to typical asthma. However, the development of CVA into asthma or normal mechanism is still lack of relevant reports. Based on this, we analyzed the differentially expressed genes of CVA, peripheral blood mononuclear cells (PBMCs) and peripheral blood mononuclear cells (PBMCs) of patients with typical asthma and normal subjects. The aim of this study was to find out the gene expression profiles of CVA different from those of typical asthmatic and normal PBMCs, so as to elucidate the pathogenesis of asthma and the possible mechanism of CVA. Methods: the peripheral venous blood samples were collected from 5 patients with CVA and 5 patients with typical asthma in each group. PBMCs were isolated and total RNAs were extracted by Trizol method, and then detected by Agilent 4 脳 44K human genome oligo microarray. After cluster analysis and gene Ontology functional analysis, 10 differentially expressed and functionally related genes were selected as candidate genes. Results: compared with CVA group, 202 differentially expressed genes were found in asthmatic group, 119 differentially expressed genes in normal group and 779 differentially expressed genes in normal group compared with CVA group. The functional analysis of Gene Ontology showed that the differentially expressed genes were mainly related to epigenetics, antigen processing and expression. Screening 10 candidate genes, 鍒嗗埆鏄疕DC,EGR1,DEFA4,LTF,G0S2,IL4,TFF3,FCER1A,CAMP,CTSG.Real-Time PCR楠岃瘉缁撴灉涓庡熀鍥犺姱鐗囩粨鏋滀笉瀹屽叏涓，

本文编号：2012289