發(fā)布時(shí)間：2019-04-04 20:28

【摘要】：目的:本研究通過對(duì)Toll樣受體4(TLR 4)基因上16個(gè)SNP位點(diǎn)進(jìn)行基因型檢測(cè)和聯(lián)合分析,探討TLR 4基因單核苷酸多態(tài)性(SNP)與類風(fēng)濕性關(guān)節(jié)炎(RA)發(fā)病的相關(guān)性。方法:1、收集福建泉州地區(qū)RA患者135例和正常對(duì)照160例外周血標(biāo)本作為研究對(duì)象,采用全血等位基因特異性擴(kuò)增(AS-PCR)技術(shù)檢測(cè)TLR4基因上16個(gè)SNP位點(diǎn)的基因型;2、采用病例對(duì)照法,借助SPSS17.0軟件、SHEsis等在線分析工具,以卡方檢驗(yàn)、t檢驗(yàn)、logistic回歸分析等方法對(duì)統(tǒng)計(jì)數(shù)據(jù)進(jìn)行分析,包括RA病例組與正常對(duì)照組間16個(gè)SNP位點(diǎn)Hardy-Weinberg平衡檢驗(yàn)、基因型和等位基因與RA的關(guān)聯(lián)分析、位點(diǎn)間的連鎖不平衡(LD)分析、單倍型和單倍型塊的分析等。結(jié)果:1、單個(gè)SNP位點(diǎn)的研究:以性別、年齡等因素對(duì)分析結(jié)果進(jìn)行校正后,位點(diǎn)rs7873784、rs7037117、rs10116253和rs10759930在RA患者組與正常對(duì)照組之間的差異具有統(tǒng)計(jì)學(xué)意義,結(jié)果如下:(1)rs7873784和rs7037117位于基因的3’末端,具有高致病風(fēng)險(xiǎn)的基因型均為GG型(P=0.011,OR=9.495,95%CI=1.169-77.131;P=0.009,OR=3.141,95%CI=1.285-7.681);分析結(jié)果未顯示前者等位基因C會(huì)增加個(gè)體的患病風(fēng)險(xiǎn),后者等位基因G具有較高致病風(fēng)險(xiǎn)(A vs.G,P=0.005,OR=1.708,95%CI=1.175-2.484);(2)rs10116253和rs10759930位于基因的5’末端,具有高致病風(fēng)險(xiǎn)的基因型均為CC型(P=0.015,OR=2.230,95%CI=1.162-4.280;P=0.035,OR=2.142,95%CI=1.050-4.371);二者等位基因C均具有較高的治病風(fēng)險(xiǎn)(T vs.C,P=0.030,OR=1.439,95%CI=1.036-2.000;T vs.C,P=0.027,OR=1.446,95%CI=1.042-2.008);2、多個(gè)SNP位點(diǎn)的研究:分別對(duì)RA病例組、正常對(duì)照組進(jìn)行LD分析,結(jié)合與RA有相關(guān)性的位點(diǎn),D’值分析中,rs7873784與rs10759930位點(diǎn)在RA組中LD較低(RA組與正常組D’值分別為0.68和0.92),r2值分析中,rs10116253和rs10759930位點(diǎn)在RA組中LD較低(RA組與正常組r2值分別為0.59和0.90);單倍型分析中,單倍體型AAGGCATTACGACGGC*(P=0.042,OR=0.530,95%CI=0.285-0.984)和AGGGCATTACGACGGC*(P=0.036,OR=1.897,95%CI=1.035-3.478)在RA病例組和正常對(duì)照組之間的差異具有統(tǒng)計(jì)學(xué)意義,各等位基因與位點(diǎn)對(duì)應(yīng)分析,其中rs7037117位點(diǎn)與RA的關(guān)系最為密切。結(jié)論:1、本研究中選取的16個(gè)SNP位點(diǎn),經(jīng)過校正有4個(gè)與RA的發(fā)病有關(guān),根據(jù)位點(diǎn)所在位置,提示基因上不同位置的SNP由于功能的不同,在RA的發(fā)病中存在差異;2、本研究中通過將16個(gè)SNP位點(diǎn)聯(lián)合分析,分析結(jié)果提示兩組的LD關(guān)系存在差異;單倍型分析中找出了這些SNP位點(diǎn)中與RA的發(fā)病關(guān)系最為密切的位點(diǎn),提示多個(gè)位點(diǎn)聯(lián)合分析的方法較單個(gè)位點(diǎn)的研究更為系統(tǒng)、實(shí)用。
[Abstract]:Aim: to investigate the association between TLR 4 gene single nucleotide polymorphism (SNP) and rheumatoid arthritis (RA) (RA) by genotyping and combined analysis of 16 SNP loci on Toll like receptor 4 (TLR 4) gene. Methods: 1. Blood samples were collected from 135 RA patients and 160 normal controls in Quanzhou, Fujian Province. The genotypes of 16 SNP loci on TLR4 gene were detected by whole blood allele-specific amplification (AS-PCR). 2. The statistical data were analyzed by Chi-square test, t-test, logistic regression analysis and other on-line analysis tools such as SPSS17.0 software, SHEsis and so on, using case-control method. Hardy-Weinberg equilibrium test of 16 SNP loci, association analysis of genotype and allele with RA, linkage disequilibrium (LD) analysis among loci, haplotype and haplotype block analysis of 16 SNP loci in RA group and normal control group were included. Results: 1. The study of single SNP locus: the difference of rs7873784,rs7037117,rs10116253 and rs10759930 between RA group and normal control group was statistically significant after correcting the analysis results by sex, age and other factors. The results were as follows: (1) rs7873784 and rs7037117 were located at the 3 'end of the gene, and the genotypes with high risk of disease were GG genotype (P = 0.011, OR 9.495, 95% CI = 1.169 * 77.131; P = 0.011, OR = 9.495,95% CI = 1.169 脳 77.131). (P) 0.009, OR 3.141, 95% CI 1.285 / 7.681); The results did not show that the former allele C increased the risk of disease, while the latter allele G had a higher risk of disease (A vs.G,P=0.005,OR=1.708,95%CI=1.175-2.484). (2) both rs10116253 and rs10759930 were located at the 5 'end of the gene, and the genotype with high risk of disease was CC genotype (P < 0.015, OR 2.230, 95% CI 1.162 / 4.280, OR 2.142, 95% CI 1.050 / 4.371), and the genotype with high risk of disease was CC genotype (P = 0.015, OR = 2.230, 95% CI = 1.162 / 4.280, OR = 2.142,95% CI = 1.050). Both allele C had a higher risk of treatment (T vs.C,P=0.030,OR=1.439,95%CI=1.036-2.000;T vs.C,P=0.027,OR=1.446,95%CI=1.042-2.008). 2, study of multiple SNP loci: LD analysis was carried out in RA case group and normal control group, combined with RA-related loci, D 'value analysis was carried out. LD of rs7873784 and rs10759930 loci were lower in RA group (D 'values of RA group and normal group were 0.68 and 0.92, respectively). In R2 analysis, LD of rs10116253 and rs10759930 loci were lower in RA group (R2 values of RA group and normal group were 0.59 and 0.90, respectively). In haplotype analysis, the difference between haplotype AAGGCATTACGACGGC* (P = 0.042, OR 0.530, 95% CI = 0.285 / 0.984) and AGGGCATTACGACGGC* (P = 0.036, OR = 1.897, 95% CI = 1.035 / 3.478) was statistically significant between RA patients and normal controls. The relationship between rs7037117 locus and RA was the most closely related to the corresponding analysis of alleles and loci. Conclusion: (1) of the 16 SNP loci selected in this study, 4 were correlated with the pathogenesis of RA after correction. According to the location of the locus, it was suggested that there were differences in the pathogenesis of RA due to the different functions of the SNP sites in the gene. 2. In this study, 16 SNP loci were analyzed and the results showed that there were differences in LD relationship between the two groups. In haplotype analysis, the most closely related loci of these SNP loci were found to be related to the pathogenesis of RA, suggesting that the method of joint analysis of multiple loci is more systematic and practical than that of single locus.
【學(xué)位授予單位】：華僑大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R593.22

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