miRNAs、MTHFR基因多态性与中国汉族人群缺血性脑卒中的关联研究

发布时间：2018-03-24 03:26

本文选题：miRNAs　切入点：MTHFR　出处：《青岛大学》2016年博士论文

【摘要】：背景和目的：缺血性脑卒中(Ischemic stroke, IS)是一类由基因和环境因素共同作用而导致的复杂性疾病。MicroRNAs (miRNAs)和亚甲基四氢叶酸还原酶(methylenetetrahydro-folate reductase, MTHFR)在缺血性脑卒中的发生和发展中起到重要作用。MicroRNAs和MTHFR基因单核苷酸多态性(single nucleotide polymorphisms, SNPs)与人类很多疾病易感性有关。近年的研究表明,miR-146aCG(rs2910164)、miR-196a2TC (rs11614913)、 miR-499 AG (rs3746444)和miR-149TC (rs2292832)已经被发现与多种疾病发病易感性有关；然而,它们对缺血性脑卒中发病易感性影响的研究报道很少,与亚型关联研究更少,且结果不一。MTHFR 3'-非翻译区(3'-untranslated region,3'-UTR) SNPs能够改变基因表达,影响与miRNAs结合,增加疾病发生风险,而其3'-UTR(rs4846049) GT多态性与缺血性脑卒中的关联及其与miRNAs基因-基因交互作用,目前尚未检索到相关报道。本研究探讨miRNAs、MTHFR基因多态性与中国汉族人群缺血性脑卒中的关联性。方法：本研究采用病例-对照设计,随机选取符合研究标准的396名缺血性脑卒中病人为病例组,对照组为378名健康人群。按照TOAST分型标准分为：大动脉粥样硬化型(large artery atherosclerosis, LAA)和小动脉闭塞型(small artery occlusion, SAO),进行亚组分析。采用聚合酶链反应-限制性片段长度多态性(Pol ym erase chain reaction-Restriction fragment length polymorphism, PCR-RFLP)和直接测序法对miR-146aCG(rs2910164)、miR-196a2 TC (rs11614913)、miR-499 AG(rs3746444)、miR-149 TC (rs2292832)和MTHFR GT(rs4846049)五个SNPs在病例组及对照组进行基因分型分析。采用拟合优度卡方检验分析Hardy-Weinberg遗传平衡(Hardy-Weinberg equilibrium, HWE);采用logistic回归模型分析SNPs及其与传统危险因素(吸烟、饮酒、糖尿病、高血压病)交互作用对缺血性脑卒中的影响；采用多因子降维法(multifactor dimensionality reduction, MDR)分析基因-基因交互作用对缺血性脑卒中的影响；采用广义线性模型分析与缺血性脑卒中相关SNPs对血脂的影响。结果：1、所有研究对象每个SNP位点基因型分布均符合Hardy-Weinberg平衡(P0.05),表示所选取样本具有人群代表性,适合行遗传学分析。2、miR-146 a CG (rs2910164) GG基因型可显著增加缺血性脑卒中发病风险(OR=1.86,95% CI:1.19-2.88,P=0.006);显性模型示：CG+GG基因型较CC基因型可增加39%的发病风险(OR=1.39,95% CI:1.04-1.86,P=0.027);隐性模型示：GG基因型较CC+CG基因型可增加62%的发病风险(OR=1.62,95% CI:1.08-2.42, P=0.020);加性模型示：每增加1个G等位基因,其发病风险增加34%(OR=1.34,95% CI:1.09-1.65, P=0.006);与C等位基因相比,G等位基因可显著增加缺血性脑卒中发病风险(OR=1.33,95% CI:1.09-1.64,P=0.006)。亚组分析示,rs2910164 CG可显著增加LAA型发病风险(P0.05),仅在校正传统危险因素后,GG基因型可增加SAO型发病风险(P0.05)。3、miR-149 TC (rs2292832) CC基因型可显著增加缺血性脑卒中的发病风险(OR=2.00,95% CI:1.22-3.28, P=0.006);显性模型示：TC+CC基因型较TT基因型可显著增加42%的发病风险(OR=1.42,95% CI:1.07-1.88, P=0.017);隐性模型示：CC基因型较TT+TC基因型可增加75%的发病风险(OR=1.75,95% CI:1.09-2.82, P=0.020);加性模型示：每增加1个C等位基因,其发病风险增加37%(OR=1.37,95% CI:1.11-1.7, P=0.004);与T等位基因相比,C等位基因可显著增加缺血性脑卒中发病风险(OR=1.37,95% CI:1.11-1.7,P=0.004)。亚组分析示,rs2292832 TC可显著增加LAA型发病风险(P0.05),未发现与SAO型发病风险显著相关性(P0.05)4、MTHFR GT (rs4846049) TT基因型可显著增加缺血性脑卒中发病风险(OR=2.87,95% CI:1.43-5.76, P=0.003);显性模型示：GT+TT基因型携带者发病风险是GG基因型的1.71倍(OR=1.71,95% CI:1.28-2.28, P0.001);隐性模型示：TT基因型携带者发病风险是GG+GT基因型的2.41倍(OR=2.41,95% CI:1.21-4.8, P=0.012);加性模型示：每增加一个T等位基因,其发病风险增加63%(OR=1.63,95% CI:1.28-2.08, P0.001);与G等位基因相比,T等位基因可显著其发病风险(OR=1.62,95% CI:1.28-2.06, P0.001)。亚组分析示,rs4846049 GT可显著增加LAA型发病风险(P0.05)；也可增加SAO型发病风险(P0.05),但校正传统危险因素后,未达到显著统计学意义(P0.05)5、未发现miR-196a2 TC (rs11614913)和miR-499 AG (rs3746444)多态性与缺血性脑卒中发病风险有显著相关性(P0.05)；亚组分析也未发现两基因位点多态性与LAA型和SAO型缺血性脑卒中的发病风险有显著关联性(P0.05)6、未发现miR-146a CG (rs2910164)、miR-149 TC (rs2292832)和MTHFR GT(rs4846049)与吸烟、饮酒、高血压病、糖尿病危险因素交互作用有显著统计学意义(P0.05)。7、miR-196a2 TC (rs11614913)、miR-499 AG (rs3746444)和MTHFR GT (rs4846049)有基因-基因交互作用(P0.05)。8、miR-149 TC (rs2292832)和MTHFR GT (rs4846049)与总胆固醇水平升高显著相关(P0.05),未发现miR-146a CG (rs2910164)多态性位点与血脂水平有显著关联性(P0.05)。结论：1、miR-146a CG (rs2910164)和miR-149 TC (rs2292832)多态性可能与中国汉族人群缺血性脑卒中发病易感性增加有关；其多态性可能主要增加LAA型缺血性脑卒中发病风险。2、我们首次发现MTHFR GT (rs4846049)多态性可能与中国汉族人群缺血性脑卒中发病易感性增加有关,可能主要增加LAA型缺血性脑卒中发病易感性。3、miR-196a2 TC (rs11614913)和miR-499 AG (rs3746444)多态性可能与中国汉族人群缺血性脑卒中发病风险无相关性。4、miR-146a CG (rs2910164)、miR-149 TC (rs2292832)和MTHFR GT (rs4846049)多态性影响中国人群缺血性脑卒中的发病易感性,可能不是通过与吸烟、饮酒、高血压病、糖尿病存在交互作用所致。5、miR-196a2 TC(rs11614913)、miR-499 AG (rs3746444)和MTHFR GT(rs4846049)之间可能存在基因-基因交互作用,影响中国汉族人群缺血性脑卒中的发病易感性。6、miR-149 TC (rs2292832)和MTHFR GT (rs4846049)多态性增加中国汉族人群缺血性脑卒中发病风险,可能与影响血脂代谢水平有关。背景和目的：亚甲基四氢叶酸还原酶(methylenetetrahydrofolate reductase,MTHFR)基因C677T多态性与缺血性脑卒中之间的关系已被广泛研究；然而,在不同的人群中结果是不一致的,特别是在中国人群中。综合评价MTHFR基因C677T多态性与缺血性脑卒中之间关联性是非常有必要的。因此,我们的研究目的是进一步去评估MTHFR基因C677T多态性与中国人群缺血性脑卒中发病风险的相关性。方法：我们从Pubmed、OVID、Embase、万方数据库、相关期刊论文(CNKI)、重庆维普数据库(VIP)和中国生物医学文献数据库(CBM),收集了所有截至到2014年8月的相关研究。汇总分析采用Stata12.0统计软件,利用OR及其95% CI评价基因多态性与缺血性脑卒中的关联强度,采用Q-检验和I2检验评价各文献之间异质性。采用不同的基因效应模型、亚组分析、发表偏倚和敏感性分析,来提高分析结果的可靠性。结果：汇总分析结果显示,MTHFR基因C677T多态性在总人群六个基因模型均存在显著关联性(加性模型OR=1.34,95%CI:1.17-1.54,P0.001;显性模型：OR=1.44,95%CI:1.26-1.64,P0.001；隐性模型：OR=1.45,95%CI:1.15-1.83,P=0.001;杂合子模型：OR=1.35,95%CI:1.18-1.55,P0.001;纯合子模型：OR=1.80,95%CI:1.34-2.41,P0.001;等位基因模型：OR=1.34,95%CI:1.17-1.53,P0.001);亚组分析中也得到同样的结果。敏感性分析显示,此分析中的各项研究具有高质量性。结论：MTHFR基因C677T多态性与中国人群缺血性脑卒中之间存在显著关联性,T等位基因可能是中国人群缺血性脑卒中一个发病危险因素。
[Abstract]:Background and purpose: ischemic stroke (Ischemic stroke IS) is a kind of gene and environmental factors lead to the complexity of.MicroRNAs disease (miRNAs) and methylenetetrahydrofolate reductase (methylenetetrahydro-folate, reductase, MTHFR) the important role of.MicroRNAs and MTHFR gene single nucleotide polymorphism plays in the occurrence and development of ischemic stroke (single nucleotide polymorphisms, SNPs) is related to many human disease susceptibility. Recent studies show that miR-146aCG (rs2910164), miR-196a2TC (rs11614913), miR-499 AG (rs3746444) and miR-149TC (rs2292832) has been found to be associated with susceptibility to various diseases; however, they reported that influence susceptibility to ischemic stroke is rare, and Asia association study is less, and the result is not a.MTHFR 3'- translation (3'-untranslated region, 3'-UTR District SNPs) can change the gene expression and effect of miRNAs combination, increase the risk, and the 3'-UTR (rs4846049) GT polymorphism and ischemic stroke and miRNAs gene gene interaction, has not yet retrieved relevant reports. This study investigated the miRNAs relationship between MTHFR gene polymorphism and ischemic stroke Chinese Han population. Methods: This study used a case-control design, randomly selected 396 ischemic stroke patients met the study criteria for the case group, the control group of 378 healthy people. According to the TOAST classification standard is divided into: large artery atherosclerosis (large artery, atherosclerosis, LAA) and artery occlusion (small type small artery occlusion, SAO), subgroup analysis. Using polymerase chain reaction restriction fragment length polymorphism (Pol YM erase chain reaction-Restriction fragment length polymorphism, PCR-RFLP) and direct sequencing of miR-146aCG (rs2910164), miR-196a2 TC (rs11614913), miR-499 AG (rs3746444), miR-149 TC (rs2292832) and MTHFR GT (rs4846049) five SNPs genotyping analyses were performed in case group and control group. Using the chi square goodness of fit test analysis of Hardy-Weinberg genetic equilibrium (Hardy-Weinberg equilibrium, HWE); logistic regression model was used to analyze SNPs and its risk factors and the traditional (smoking, drinking, diabetes, hypertension) interaction effects on ischemic stroke; using multifactor dimensionality reduction (multifactor dimensionality, reduction, MDR) analysis of gene gene interactions on ischemic stroke; the influence of generalized linear model of ischemic stroke and SNPs on blood lipids. Results: 1, all of the subjects of each SNP genotype distribution was in accordance with Hardy-Weinberg balance (P0. 05), suggested that the selected samples could represent the population genetics analysis, suitable for.2, miR-146 a CG (rs2910164) GG genotype significantly increased the risk of ischemic stroke (OR=1.86,95% CI:1.19-2.88, P=0.006); dominant model: CG+GG genotype compared with CC genotype could increase the risk of 39% (OR=1.39,95% CI:1.04-1.86, P=0.027); recessive model: GG genotype compared with CC+CG genotype could increase the risk of 62% (OR=1.62,95%, CI:1.08-2.42, P=0.020); the additive model showed that every increase of 1 G alleles, the risk increased by 34% (OR=1.34,95%, CI:1.09-1.65, P=0.006); compared with C allele, G allele can significantly increase the risk of ischemic stroke (OR=1.33,95% CI:1.09-1.64, P=0.006). Subgroup analysis showed that rs2910164 CG significantly increased the risk of type LAA (P0.05), only after adjustment for conventional risk factors, the GG genotype can increase With the risk of type SAO (P0.05).3 miR-149 TC (rs2292832) CC genotype significantly increased the risk of ischemic stroke (OR=2.00,95% CI:1.22-3.28, P=0.006); dominant model: TC+CC genotype compared to TT genotype significantly increased the risk of 42% (OR= 1.42,95%, CI:1.07-1.88, P=0.017); the recessive model shows: compared with the CC genotype TT+TC genotype could increase the risk of 75% (OR=1.75,95%, CI:1.09-2.82, P=0.020); the additive model showed that every increase of 1 C alleles, the risk increased by 37% (OR=1.37,95% CI:1.11-1.7, P=0.004); compared with T allele, C allele significantly increased the risk of ischemic stroke stroke (OR=1.37,95% CI:1.11-1.7, P=0.004). Subgroup analysis showed that rs2292832 TC significantly increased the risk of type LAA (P0.05), and the risk of type SAO not found significant correlation (P0.05 4), MTHFR GT (rs4846049) TT genotype Can significantly increase the risk of ischemic stroke (OR=2.87,95% CI:1.43-5.76, P=0.003); the dominant model shows: the risk of GT+TT genotype carriers was 1.71 times of the GG genotype (OR=1.71,95%, CI:1.28-2.28, P0.001); the recessive model showed risk carriers of TT genotype was 2.41 times than that of GG+GT genotype (OR=2.41,95% CI:1.21-4.8, P=0.012); and the model shows: every increase of one T allele, the risk increased by 63% (OR=1.63,95% CI:1.28-2.08, P0.001); compared with G allele, T allele was the risk (OR=1.62,95% CI:1.28-2.06, P0.001). Subgroup analysis showed that rs4846049 GT significantly increased the risk of type LAA (P0.05); also can increase the risk of type SAO (P0.05), but after adjustment for conventional risk factors did not reach statistical significance (P0.05) 5, miR-196a2 was not found in TC (rs11614913) and miR-499 AG (rs3746444) - There was a significant correlation between normal and ischemic stroke risk (P0.05); subgroup analysis has not been found because of the risk of two polymorphisms and LAA type and SAO type of ischemic stroke have significant correlation (P0.05) 6, miR-146a was not found in CG (rs2910164), miR-149 TC (rs2292832) and MTHFR (GT rs4846049) and smoking, drinking, hypertension, there was significant interaction between risk factors of diabetes mellitus (P0.05).7 miR-196a2 TC (rs11614913), miR-499 AG (rs3746444) and MTHFR GT (rs4846049) gene - gene interactions (P0.05.8), miR-149 TC (rs2292832) and MTHFR GT (rs4846049) was associated with elevated levels of total cholesterol (P0.05), miR-146a was not found in CG (rs2910164) polymorphism and serum lipid levels have significant correlation (P0.05). Conclusion: 1. MiR-146a CG (rs2910164) and miR-149 TC (rs2292832) polymorphism and China Han people The group of ischemic stroke susceptibility; the polymorphism may increase LAA ischemic stroke risk.2, we first found that MTHFR GT (rs4846049) polymorphism may be associated with increased Chinese Han population of ischemic stroke may increase the susceptibility of LAA type of cerebral ischemic stroke susceptibility.3, miR-196a2 TC (rs11614913) miR-499 and AG (rs3746444) polymorphism may not be associated with.4 in Han population Chinese ischemic stroke risk, miR-146a CG (rs2910164), miR-149 TC (rs2292832) and MTHFR GT (rs4846049) Chinese population susceptibility to ischemic stroke polymorphism may not influence, with smoking, drinking, hypertension, diabetes the interaction caused by the.5 miR-196a2 TC (rs11614913), miR-499 AG (rs3746444) and MTHFR GT (rs4846049) may exist between genes The interaction effect of the susceptibility to.6 in Han population China ischemic stroke, miR-149 TC (rs2292832) and MTHFR GT (rs4846049) polymorphism in Han population of Chinese increased ischemic stroke risk may be related to lipid metabolism. Background and purpose: methylenetetrahydrofolate reductase (methylenetetrahydrofolate, reductase, MTHFR) the relationship between C677T gene polymorphism ischemic stroke has been studied extensively; however, in different populations results is inconsistent, especially in the Chinese population. A comprehensive evaluation of the association between MTHFR gene C677T polymorphism and ischemic stroke is necessary. Therefore, the aim of our study is to further evaluate the relationship between MTHFR gene C677T China population polymorphism and risk of ischemic stroke. Methods: from Pubmed, OVID, Embase, Wanfang Data Library, Chinese Journal Full-text Database (CNKI), Chongqing VIP database (VIP) and Chinese biomedical literature database (CBM), collected all the relevant research by August 2014. Analyzed by Stata12.0 statistical software, using the correlation intensity OR and evaluation of 95% CI gene polymorphism and ischemic stroke, by Q- test and I2 the testing and evaluation of the literature. The gene effect heterogeneity among different models, subgroup analysis, publication bias and sensitivity analysis, to improve the reliability of the analysis results. Results: the pooled analysis showed that MTHFR gene C677T polymorphism in model six genes total population there were significant correlation (additive model OR=1.34,95%CI:1.17-1.54, P0.001; dominant model: OR=1.44,95%CI:1.26-1.64, P0.001; OR=1.45,95%CI:1.15-1.83, P=0.001; the recessive model: heterozygous model: OR= 1.35,95%CI:1.18-1.55, P0.001; pure Zygotic model: OR=1.80,95%CI:1.34-2.41, P0.001; allele model: OR=1.34,95%CI:1.17-1.53, P0.001); also the results of the subgroup analysis. The sensitivity analysis shows that this research in the analysis of high quality. Conclusion: there was significant correlation between MTHFR gene C677T polymorphism and ischemic stroke China population, T allele Chinese gene may be a population of ischemic stroke risk factors.

【学位授予单位】：青岛大学
【学位级别】：博士
【学位授予年份】：2016
【分类号】：R743.3

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