MMPs基因启动子区5个SNPs与河南汉族群体CHF遗传易感性的关联研究

发布时间：2018-07-03 01:16

本文选题：单核苷酸多态性 + 限制性扩增片段长度多态性　；参考：《新乡医学院》2016年硕士论文

【摘要】：背景慢性充血性心力衰竭(chronic heart failure,CHF)是一种严重影响人类健康的一种疾病。现在人们正在深入探索该疾病发生的分子机制。基质金属蛋白酶(matrix metalloproteinases,MMPs)活性的增强与表达的改变,对脉管系统及心脏组织的重构产生严重的影响,造成心血管系统的损害,结果将会造成心功能恶化甚至心衰。目的研究MMPs基因启动子区5个单核苷酸多态性(single nucleotide polymorphisms,SNPs)与河南汉族群体慢性充血性心力衰竭的关系,以探讨河南汉族群体慢性充血性心力衰竭发病的分子机制。方法1、样本收集与基因组DNA提取:根据知情同意的原则,收集CHF患者160例,正常对照186例,酚/氯仿方法提取基因组DNA。2、SNPs位点检测及分型方法:采用限制性扩增片段长度多态性(polymerase chain reaction restriction fragment length polymorphism,PCR-RFLP)的方法检测MMPs基因rs2285053,rs243865,rs11568818,rs11225395及rs225207共5个单核苷酸多态性。3、统计学原理分析:将不同的每组基因型及等位基因制成Excle表格,采用病例-对照分析,运用SHEsis软件,将病例组与对照组间频数及频率分布进行比较分析、Hardy-Weinberg平衡检验、单体型推断、比值比(odd ratio,OR)及95%置信区间(confidence interval,CI)计算;并分析3个多态连锁不平衡(linkage d isequilibrium,LD)紧密程度。所有统计检验均为双侧检验,检验水准α=0.05。如果计算结果不符合统计学中的正态分布,通过平方根转换后,进行t检验。结果1、MMPs基因型分布Hardy-Weinberg平衡检验:rs2285053、rs11225395及rs225207 3个SNPs位点的基因及基因型频率在两组人群中符合Hardy-Weinberg平衡(P0.05)。rs243865位点基因型全部为CC,rs11568818位点基因型全部为AA,有可能rs243865及rs11568818在河南汉族人群中没有多态性,因此不予分析。2、MMPs基因3个SNPs与CHF之间的相关性:心衰组rs2285053位点的T等位基因频率降低(P=0.043);心衰组rs11225395位点的T等位基因的频率显著降低(P=0.004),OR=1.966[95%CI(1.236-3.129)],TT基因型频率与对照组相比也显著降低(P=0.009);rs225207的G等位基因频率显著增加(P=0.002),OR=0.499[95%CI(0.320-3.779)],GG基因型频率与对照组相比也显著增加(P=0.007)。3、MMPs基因启动区3个SNPs单体型分析:CCG单体型的频率在病例组显著高于对照组(P0.01),OR=3.436[95%CI(1.888~6.253)]。结论1、与对照组相比,心衰组rs2285053及rs11225395位点的T等位基因频率降低,rs225207位点的G等位基因及单体型CCG的频率升高;2、rs2285053及rs11225395位点的T等位基因可能会降低个体慢性充血性心力衰竭的患病风险,rs225207位点的G等位基因及单体型CCG可能会增加个体慢性充血性心力衰竭的患病风险。
[Abstract]:Background chronic congestive heart failure (chronic heart) is a serious disease affecting human health. The molecular mechanism of the disease is now being explored in depth. The enhancement and expression of matrix metalloproteinases (MMPs) have a serious effect on the remodeling of vascular system and heart tissue, resulting in the damage of cardiovascular system, resulting in the deterioration of cardiac function and even heart failure. Objective to study the relationship between five single nucleotide polymorphisms (single nucleotide polymorphismsns) of MMPs gene promoter region and chronic congestive heart failure (CHF) in Henan Han population, and to explore the molecular mechanism of CHF in Henan Han population. Methods 1. Sample collection and genomic DNA extraction: 160 CHF patients and 186 normal controls were collected according to the principle of informed consent. Detection and typing of genomic DNA.2SNPs by phenol / chloroform method: restriction amplified fragment length polymorphism (polymerase chain reaction) was used to detect 5 single nucleotide polymorphisms (s2285053rs243865rs11568818rs11225395 and rs225207). Analysis: make different groups of genotypes and alleles into an Excle table, The frequency and frequency distribution between the case group and the control group were compared and analyzed by case-control analysis and SHEsis software. The Hardy-Weinberg equilibrium test, haplotype inference, odd ratioor and 95% confidence interval (confidence intervalval CI) were calculated. The tightness of three polymorphic linkage disequilibrium (linkage d isequilibrium LD) was analyzed. All the statistical tests were bilateral, and the test level was 0. 05. If the calculated results do not conform to the normal distribution in statistics, the t-test is carried out after the square root conversion. Results 1. Hardy-Weinberg equilibrium test showed that the gene and genotype frequencies of the three SNPs loci in the two groups were in accordance with the Hardy-Weinberg equilibrium (P0.05) .rs243865 alleles were all CCRs11568818 alleles. It may be that rs243865 and rs11568818 are all AAs in Han Han, Henan Province. There is no polymorphism in the ethnic group, Therefore, the correlation between three SNPs of MMPs gene and CHF was not analyzed: the frequency of T allele at rs2285053 locus in heart failure group was decreased (P0.043), and the frequency of T allele at rs11225395 locus in heart failure group was significantly lower than that in control group (P0. 004), and the frequency of TT genotype was also significantly lower than that in control group (P0. 004). The G allele frequency of Rs225207 decreased significantly (P0. 002) ORO 0.499 [95CI (0.320-3.779)] GG genotype frequency was also significantly increased compared with the control group (P0. 007). 3 SNPs haplotype analysis of three SNPs haplotypes of MMPs gene promoter region in the case group was significantly higher than that in the control group (P0.01) OR3.436 [95CI (1. 888m6.253)]. Conclusion 1.Compared with the control group, Lower T allele frequency at rs2285053 and rs11225395 loci in heart failure group A decrease in G allele at rs225207 locus and an increase in haplotype CCG allele T alleles at rs2285053 and rs11225395 locus may reduce the risk of chronic congestive heart failure (CHF) in individuals with chronic congestive heart failure (CHF). G allele and haplotype CCG may increase the risk of chronic congestive heart failure.
【学位授予单位】：新乡医学院
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R541.6

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