系统性红斑狼疮患者外周血单个核细胞IL-28RA基因mRNA表达研究

发布时间：2018-05-31 22:26

本文选题：红斑狼疮/系统性 + 外周血单个核细胞　；参考：《安徽医科大学》2015年硕士论文

【摘要】：研究背景系统性红斑狼疮(SLE)是一种多发生于青年女性,可累及多器官、多系统,以自身抗体产生和免疫复合物沉积为病理特征的自身免疫性结缔组织疾病。SLE发病机制到现在尚未完全明确,大量研究显示,遗传、内分泌、感染、免疫异常和一些环境因素与本病的发生有关,其中,遗传因素在SLE的发病中起到重要作用。过去二十多年,许多研究小组利用遗传连锁和关联研究,发现了大量SLE的易感基因,如HLA,STAT4,IRF5,ITGAM和PTPN22等。随着Human Haplotype Map(Hap Map)的完成以及高效廉价的高通量基因分型实验技术的应用,2008年以来,利用全基因组关联分析(Genome-wide association study,GWAS)在对不同种族人的SLE患者进行研究时发现了40多个易感基因/位点。其中,2013年,李扬等以GWAS数据库(7260个银屑病病例和9842个对照以及2207个SLE病例和9842个对照)为基础,通过GWAS分析,发现了银屑病与SLE共同的易感基因位点(NFKBIA和IL-28RA),其中,IL-28RA(全称为Interleukin-28 receptor,alpha,又名IFNLR1;LCR2;CRF2/12;IL-28R1)基因的单个核苷酸多态性(single nucleotide polymorphism,SNP)rs4649203与SLE易感性显著相关(P=9.90×10-09)。IL-28RA基因编码的蛋白属于II型细胞因子受体家族,此蛋白同IL-10受体β(Interleukin-10receptor,beta,IL-10RB)组成一个受体复合物,此受体复合物与III型IFN的3个细胞因子(IL-28A、IL-28B、IL-29)相互作用,细胞高表达此受体复合体后导致细胞对于IL-28A、IL-29的反应增强,而对于IL-28B的反应减弱,通过刺激JAK-STAT信号通路,从而在细胞的增殖、分化、凋亡以及免疫调节等多种生物学过程发挥作用。IL-28RA基因与银屑病、SLE、慢性丙型肝炎、多发性硬化症、过敏性鼻炎等疾病有关。本研究通过运用实时荧光定量PCR技术,测定IL-28RA基因的m RNA在SLE患者以及正常对照中的表达情况,来探讨IL-28RA基因在SLE发病中的作用及其可能作用机制。目的探讨SLE患者与健康对照的外周血学单个核细胞(PBMCs)中IL-28RA基因m RNA表达情况及差异,了解IL-28RA基因m RNA表达与SLE活动度指数(SLEDAI)及易感位点SNP rs4649203的相关性。方法收集SLE患者62例,正常对照69例,从外周血单个核细胞(PBMCs)中提取出m RNA,然后逆转录为complementary DNA(c DNA),加入相应的荧光染料、引物,利用ABI 7900HT高通量荧光定量PCR技术检测外周血单个核细胞中IL-28RA基因m RNA的表达情况;从研究对象外周血提取DNA,浓度标准化后,经Sequenom Mass Array技术对所有研究对象DNA样本进行SNP rs4649203位点的基因分型;收集整理所有实验数据,用SPSS20.0软件进行数据统计分析。结果SLE患者IL-28RA基因m RNA表达水平高于正常对照组,差异具有统计学意义(P0.001);IL-28RA基因m RNA表达水平与SLEDAI无相关性(r=0.045,P=0.726);IL-28RA基因m RNA表达水平与SNP rs4649203无相关性(P0.05)。结论IL-28RA基因表达差异提示其在SLE发病机制中可能起到某种重要的调节作用。
[Abstract]:Background systemic lupus erythematosus (SLE) is an autoimmune connective tissue disease (.SLE) pathogenesis of multiple organs and multiple systems involving multiple organs and multiple systems. The pathogenesis of autoimmune connective tissue disease is not completely clear. A large number of studies have shown that heredity, endocrinology, infection, immune abnormality, and abnormality. Some environmental factors are related to the occurrence of this disease, among which genetic factors play an important role in the pathogenesis of SLE. In the past more than 20 years, many research teams have discovered a large number of SLE susceptible genes, such as HLA, STAT4, IRF5, ITGAM and PTPN22, using genetic linkage and Association studies. With the completion of Human Haplotype Map (Hap Map) and high validity The application of the high throughput genotyping test technique, since 2008, more than 40 susceptible genes / loci were found by Genome-wide association study (GWAS) in the study of SLE patients of different ethnic groups. In 2013, Li Yang was used as a GWAS database (7260 psoriasis cases and 9842 controls, as well as 9842 controls, and) On the basis of 2207 SLE cases and 9842 controls, the common allelic loci (NFKBIA and IL-28RA) of psoriasis and SLE (NFKBIA and IL-28RA) were found through GWAS analysis, in which the single nucleotide polymorphism of Interleukin-28 receptor, alpha, LCR2; CRF2/12; SLE susceptibility significant correlation (P=9.90 x 10-09).IL-28RA gene encoded protein belongs to the II cell factor receptor family, which is composed of a receptor complex with IL-10 receptor beta (Interleukin-10receptor, beta, IL-10RB). The receptor complex interacts with the 3 cytokines of III IFN, and the cell is highly expressed. After the body complex, the cell response to IL-28A, IL-29 is enhanced, and the response to IL-28B is weakened. The.IL-28RA gene and psoriasis, SLE, chronic hepatitis C, multiple sclerosis, allergic rhinology, and allergic rhinitis are played by stimulating the JAK-STAT signaling pathway by stimulating the proliferation, differentiation, apoptosis and immunoregulation of the cells. The purpose of this study is to determine the role of the IL-28RA gene in the pathogenesis of SLE and its possible mechanism by using real time fluorescence quantitative PCR technique to determine the expression of the m RNA of the IL-28RA gene in SLE patients and normal controls. The purpose of this study is to explore the IL-28RA of SLE patients and the healthy control of peripheral blood mononuclear cells (PBMCs) IL-28RA. The relationship between the expression of M RNA and the correlation between the expression of M RNA of the IL-28RA gene and the SLE activity index (SLEDAI) and the rs4649203 of the susceptible loci. Methods 62 cases of SLE patients and 69 normal controls were collected from the peripheral blood mononuclear cells (PBMCs), and then the reverse transcription was added to the corresponding fluorescence staining. Materials, primers, using ABI 7900HT high throughput fluorescence quantitative PCR technique to detect the expression of IL-28RA gene m RNA in peripheral blood mononuclear cells. After extracting DNA from the peripheral blood of the research object, the concentration of DNA is standardized, and Sequenom Mass Array technique is used to classify all the DNA samples of the subjects by the Sequenom Mass Array technique. All the facts are collected and collected. The data were analyzed by SPSS20.0 software. Results the expression level of IL-28RA gene m RNA in SLE patients was higher than that in the normal control group, the difference was statistically significant (P0.001), and the RNA expression level of IL-28RA gene m was not related to SLEDAI (r=0.045, P=0.726). The difference of 28RA gene expression suggests that it may play an important regulatory role in the pathogenesis of SLE.
【学位授予单位】：安徽医科大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R593.241

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