本文選題：咳嗽變異性哮喘 + 支氣管哮喘�。� 參考：《川北醫(yī)學院》2017年碩士論文

【摘要】：目的:咳嗽變異性哮喘(Cough variant asthma,CVA)是一種不典型的支氣管哮喘(Bronchial asthma,以下簡稱哮喘),因其可發(fā)展為典型哮喘,也被稱為哮喘的前驅(qū)階段,早期正確治療可部分避免由CVA向典型哮喘的進一步發(fā)展。而CVA發(fā)展成為哮喘或正常的機制目前尚缺乏相關(guān)報道�；诖�,本研究對CVA、典型哮喘以及正常人外周血單個核細胞(Peripheral Blood Mononuclear Cells,PBMCs)的差異表達基因進行分析。以期找出CVA異于典型哮喘及正常人PBMCs的基因表達譜,借此進一步闡述哮喘的發(fā)病機制及CVA的可能轉(zhuǎn)歸機制。方法:收集正常人、CVA患者及典型哮喘患者各5例,每組各例分別采集外周靜脈血3ml,分離出PBMCs,并利用Trizol法提取總RNA,再用Agilent 4×44K人類全基因組Oligo芯片進行檢測,并對其差異基因行聚類分析及Gene Ontology功能分析,隨之挑選10個差異表達顯著且功能相關(guān)的基因作為候選基因,然后利用實時熒光定量PCR(Real-Time PCR,RT-PCR)技術(shù)對候選基因進行驗證。結(jié)果:基因芯片檢測結(jié)果發(fā)現(xiàn),哮喘組與CVA組比較,有差異表達基因202個,正常組與CVA組比較,有差異表達基因119個,正常組與哮喘組比較,有差異表達基因779個,差異均在2倍以上。Gene Ontology功能分析提示差異基因主要與表觀遺傳學及抗原處理和表達等相關(guān)。篩選10個候選基因,分別是HDC、EGR1、DEFA4、LTF、G0S2、IL4、TFF3、FCER1A、CAMP、CTSG。Real-Time PCR驗證結(jié)果與基因芯片結(jié)果不完全一致。FCER1A、HDC、IL4基因在CVA組中較正常組以及哮喘組均表達上調(diào)(P0.05),其相對表達量分別為:(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基因的表達在三組間比較差異均無統(tǒng)計學意義(P0.05)。結(jié)論:1、本研究人全基因組表達芯片結(jié)果顯示CVA與典型哮喘及正常人PBMC的基因表達譜均存在明顯差異。2、Real-Time PCR驗證結(jié)果顯示,FCER1A、HDC、IL4基因在CVA組中較正常組以及哮喘組均表達上調(diào),EGR1、DEFA4、LTF、G0S2、TFF3、CAMP、CTSG基因在三組間的比較差異均無統(tǒng)計學意義。提示CVA的發(fā)病可能與FCER1A、HDC、IL4基因有關(guān),而其余7個基因可能不是影響CVA發(fā)病及轉(zhuǎn)歸的重要基因。
[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楠岃瘉緇撴灉涓庡熀鍥犺姱鐗囩粨鏋滀笉瀹屽叏涓，

本文編號：2012289