TLR4基因多态性与类风湿性关节炎发病相关性的研究

发布时间：2019-04-04 20:28

【摘要】：目的:本研究通过对Toll样受体4(TLR 4)基因上16个SNP位点进行基因型检测和联合分析,探讨TLR 4基因单核苷酸多态性(SNP)与类风湿性关节炎(RA)发病的相关性。方法:1、收集福建泉州地区RA患者135例和正常对照160例外周血标本作为研究对象,采用全血等位基因特异性扩增(AS-PCR)技术检测TLR4基因上16个SNP位点的基因型;2、采用病例对照法,借助SPSS17.0软件、SHEsis等在线分析工具,以卡方检验、t检验、logistic回归分析等方法对统计数据进行分析,包括RA病例组与正常对照组间16个SNP位点Hardy-Weinberg平衡检验、基因型和等位基因与RA的关联分析、位点间的连锁不平衡(LD)分析、单倍型和单倍型块的分析等。结果:1、单个SNP位点的研究:以性别、年龄等因素对分析结果进行校正后,位点rs7873784、rs7037117、rs10116253和rs10759930在RA患者组与正常对照组之间的差异具有统计学意义,结果如下:(1)rs7873784和rs7037117位于基因的3’末端,具有高致病风险的基因型均为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);分析结果未显示前者等位基因C会增加个体的患病风险,后者等位基因G具有较高致病风险(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,具有高致病风险的基因型均为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均具有较高的治病风险(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、多个SNP位点的研究:分别对RA病例组、正常对照组进行LD分析,结合与RA有相关性的位点,D’值分析中,rs7873784与rs10759930位点在RA组中LD较低(RA组与正常组D’值分别为0.68和0.92),r2值分析中,rs10116253和rs10759930位点在RA组中LD较低(RA组与正常组r2值分别为0.59和0.90);单倍型分析中,单倍体型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病例组和正常对照组之间的差异具有统计学意义,各等位基因与位点对应分析,其中rs7037117位点与RA的关系最为密切。结论:1、本研究中选取的16个SNP位点,经过校正有4个与RA的发病有关,根据位点所在位置,提示基因上不同位置的SNP由于功能的不同,在RA的发病中存在差异;2、本研究中通过将16个SNP位点联合分析,分析结果提示两组的LD关系存在差异;单倍型分析中找出了这些SNP位点中与RA的发病关系最为密切的位点,提示多个位点联合分析的方法较单个位点的研究更为系统、实用。
[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.
【学位授予单位】：华侨大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R593.22

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