本文選題：紅斑狼瘡/系統(tǒng)性 + 外周血單個核細(xì)胞��； 參考：《安徽醫(yī)科大學(xué)》2015年碩士論文

【摘要】：研究背景系統(tǒng)性紅斑狼瘡(SLE)是一種多發(fā)生于青年女性,可累及多器官、多系統(tǒng),以自身抗體產(chǎn)生和免疫復(fù)合物沉積為病理特征的自身免疫性結(jié)締組織疾病。SLE發(fā)病機(jī)制到現(xiàn)在尚未完全明確,大量研究顯示,遺傳、內(nèi)分泌、感染、免疫異常和一些環(huán)境因素與本病的發(fā)生有關(guān),其中,遺傳因素在SLE的發(fā)病中起到重要作用。過去二十多年,許多研究小組利用遺傳連鎖和關(guān)聯(lián)研究,發(fā)現(xiàn)了大量SLE的易感基因,如HLA,STAT4,IRF5,ITGAM和PTPN22等。隨著Human Haplotype Map(Hap Map)的完成以及高效廉價的高通量基因分型實驗技術(shù)的應(yīng)用,2008年以來,利用全基因組關(guān)聯(lián)分析(Genome-wide association study,GWAS)在對不同種族人的SLE患者進(jìn)行研究時發(fā)現(xiàn)了40多個易感基因/位點(diǎn)。其中,2013年,李揚(yáng)等以GWAS數(shù)據(jù)庫(7260個銀屑病病例和9842個對照以及2207個SLE病例和9842個對照)為基礎(chǔ),通過GWAS分析,發(fā)現(xiàn)了銀屑病與SLE共同的易感基因位點(diǎn)(NFKBIA和IL-28RA),其中,IL-28RA(全稱為Interleukin-28 receptor,alpha,又名IFNLR1;LCR2;CRF2/12;IL-28R1)基因的單個核苷酸多態(tài)性(single nucleotide polymorphism,SNP)rs4649203與SLE易感性顯著相關(guān)(P=9.90×10-09)。IL-28RA基因編碼的蛋白屬于II型細(xì)胞因子受體家族,此蛋白同IL-10受體β(Interleukin-10receptor,beta,IL-10RB)組成一個受體復(fù)合物,此受體復(fù)合物與III型IFN的3個細(xì)胞因子(IL-28A、IL-28B、IL-29)相互作用,細(xì)胞高表達(dá)此受體復(fù)合體后導(dǎo)致細(xì)胞對于IL-28A、IL-29的反應(yīng)增強(qiáng),而對于IL-28B的反應(yīng)減弱,通過刺激JAK-STAT信號通路,從而在細(xì)胞的增殖、分化、凋亡以及免疫調(diào)節(jié)等多種生物學(xué)過程發(fā)揮作用。IL-28RA基因與銀屑病、SLE、慢性丙型肝炎、多發(fā)性硬化癥、過敏性鼻炎等疾病有關(guān)。本研究通過運(yùn)用實時熒光定量PCR技術(shù),測定IL-28RA基因的m RNA在SLE患者以及正常對照中的表達(dá)情況,來探討IL-28RA基因在SLE發(fā)病中的作用及其可能作用機(jī)制。目的探討SLE患者與健康對照的外周血學(xué)單個核細(xì)胞(PBMCs)中IL-28RA基因m RNA表達(dá)情況及差異,了解IL-28RA基因m RNA表達(dá)與SLE活動度指數(shù)(SLEDAI)及易感位點(diǎn)SNP rs4649203的相關(guān)性。方法收集SLE患者62例,正常對照69例,從外周血單個核細(xì)胞(PBMCs)中提取出m RNA,然后逆轉(zhuǎn)錄為complementary DNA(c DNA),加入相應(yīng)的熒光染料、引物,利用ABI 7900HT高通量熒光定量PCR技術(shù)檢測外周血單個核細(xì)胞中IL-28RA基因m RNA的表達(dá)情況;從研究對象外周血提取DNA,濃度標(biāo)準(zhǔn)化后,經(jīng)Sequenom Mass Array技術(shù)對所有研究對象DNA樣本進(jìn)行SNP rs4649203位點(diǎn)的基因分型;收集整理所有實驗數(shù)據(jù),用SPSS20.0軟件進(jìn)行數(shù)據(jù)統(tǒng)計分析。結(jié)果SLE患者IL-28RA基因m RNA表達(dá)水平高于正常對照組,差異具有統(tǒng)計學(xué)意義(P0.001);IL-28RA基因m RNA表達(dá)水平與SLEDAI無相關(guān)性(r=0.045,P=0.726);IL-28RA基因m RNA表達(dá)水平與SNP rs4649203無相關(guān)性(P0.05)。結(jié)論IL-28RA基因表達(dá)差異提示其在SLE發(fā)病機(jī)制中可能起到某種重要的調(diào)節(jié)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.
【學(xué)位授予單位】：安徽醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2015
【分類號】：R593.241

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